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Research Article | Open Access
Prokaryotic Expression of α-13 Giardin Gene and Its Intracellular Localization in Giardia lamblia
To study prokaryotic expression and subcellular localization of α-13 giardin in Giardia lamblia trophozoites, α-13 giardin gene was amplified and cloned into prokaryotic expression vector pET-28a(+). The positive recombinant plasmid was transformed into E. coli BL21(DE3) for expression by using IPTG and autoinduction expression system (ZYM-5052). The target protein was validated by SDS-PAGE and Western blotting and purified by Ni-NTA Resin. Rabbits were immunized with purified fusion proteins for preparation of polyclonal antibody; then the intracellular location of α-13 giardin was determined by fluorescence immunoassay. The results showed that the length of α-13 giardin gene was 1038 bp, encoding a polypeptide of 345 amino acids. The expressed product was a fusion protein with about 40 kDa largely present in soluble form. The target protein accounted for 21.0% of total proteins after being induced with IPTG, while it accounted for 28.8% with ZYM-5052. The anti-α13-giardin polyclonal antibody possessed good antigenic specificity as well as excellent binding activity with recombinant α-13 giardin. Immunofluorescence assays revealed that α-13 giardin was localized in the cytoplasm of G. lamblia trophozoite, suggesting that it is a cytoplasm-associated protein. The present study may lay a foundation for further functional research on α-13 giardin of G. lamblia.
1. Introduction
Giardia lamblia (syn. G. intestinalis, G. duodenalis) is an important zoonotic parasite that infects many mammals, including cats, dogs, and humans. G. lamblia assemblage A and B infection in individuals can lead to abdominal cramps, acute or chronic diarrhea, and malabsorption [1]. Since the 1970s, the disease has been epidemic or outbreak all over the world; more than 280 million of human symptomatic infections each year only in Africa, Asia, and America are estimated by the World Health Organization (WHO) [2, 3].
G. lamblia has two stages of life cycle: an infective, immotile cyst and a vegetative, motile trophozoite. The latter (trophozoite) has a pear-shaped body, which is ventrally flattened and bilaterally symmetric. Besides, it has two adhesive disks on the ventral surface, a microtubular median body, two nuclei, and four pairs of flagella: anterior, ventral, lateroposterior, and caudal [4]. Giardins, unique component of the cytoskeleton of Giardia, have been identified as four major classes: alpha (α), beta (β), gamma (γ), and delta (δ) [5]. Among them, 21 α-giardins numbered from α-1 to α-19 giardin (α-7 giardin subdivided as α-7.1, α-7.2, and α-7.3) have been discovered; their relative molecular masses are about 29~38 kDa, with common characteristic of calcium dependency membrane bound proteins containing acidic phospholipid. They are often associated with the plasma membrane and membrane systems and participate in the movement of the cytoskeleton and signal transduction in the cell, as well as regulating the growth and proliferation of cells and forming the atypical Ca2+ channels [6, 7]. In addition, some α-giardins also participate in the encystation and excystation process of Giardia cysts, preventing their leakage of the cell membrane and disintegration of the cellular structure under the environment of high intestinal bile salts [6].
In recent years, numerous researches on the subcellular localization of α-giardins have been reported. Weiland et al. [6] identified 14 coding genes for α-giardins (α-4 to α-6, α-8 to α-13, and α-15 to α-19) in G. lamblia. The localization experiments showed that a few α-giardins (α-3, α-5, and α-17) are localized in the adhesive disc; α-15 and α-16 giardins had a patchy distribution along the plasma membrane, while the others (α-1, α-2, α-6, α-7.2, α-7.3, α-9, α-10, and α-14) were mainly distributed in plasma membrane, adhesive disc, flagella, or cytoplasm. Alpha 1 (α-1) giardin was identified mainly present in plasma membrane in strains of assemblages A but also in assemblages B and E [8]. Alpha 8 (α-8) and α-4 giardins were found to be mainly located on the flagella and plasma membrane (the former) and flagella by specific polypeptide, respectively [9, 10]. The immunofluorescence assay revealed that α-11 giardin is mainly localized in the plasma membrane and basal body of the anterior flagella of G. lamblia trophozoite [11]. Recently we have successes on sequence analysis and prokaryotic expression of G. lambliaα-18 giardin gene [12]. And we founded that α18- and α12-giardin proteins were mainly localized in the flagella and cytoplasm of trophozoites, respectively [13]. However, there are no reports about α-13 giardin to date.
In this research work, we proceed to provide new reports on prokaryotic expression of α-13 giardin gene and its intracellular localization in G. lamblia assemblage A to lay a foundation for further functional study of α-13 giardin.
2. Materials and Methods
2.1. Ethics
Animal experiments involved in this study were treated in accordance with the guidelines of the South China Agricultural University Animal Care and Use Committee, which operates under the Animal Welfare Law and Regulations of the Department of Health and Human Services. The South China Agricultural University Animal Care and Use Committee has approved all protocols of this study.
2.2. Cell Culture
Trophozoites of G. lamblia strain WBC6 (ATCC 50803) were cultured in Keister's modified TYI-S-33 medium [14] at 37°C, which was placed in 10 mL glass tubes. The cultures were decanted into 15 mL centrifuge tubes at the end of the logarithmic phase (after about 3 days), and trophozoites were harvested at 5400 ×g for 10 min at 4°C.
2.3. PCR Amplification
Total DNA was extracted from trophozoites using a commercial DNA Extraction Kit (Promega, Madison, USA) and quantified by spectrophotometry at 260 nm. Two primers specific to α-13 giardin gene, A13E (5′-CGG GAT CCA TGC CTG TTC TGA CCC C-3′) with Bam HI restriction site (underlined) and A13F (5′-CCC AAG CTT CTA ATC CAC ATC CCA GAG CC-3′) with Hind III restriction site (underlined), were designed based on the G. lamblia nucleotide sequence (GL50803_1076). The predicted amplification fragment was 1038 bp. A 25 μL of PCR reaction mix contained 17.3 μL of ddH2O, 2.5 μL of 10x PCR Buffer (Mg2+ plus), 2.5 μL dNTP, 0.2 μL of upstream and downstream primers (50 pmol/L), 0.3 μL of ExTaq DNA polymerase, and 2 μL of DNA sample. The thermocycler program consisted of 95°C for 5 min, followed by 35 cycles of 94°C for 30 s, 55.2°C for 30 s, and 72°C for 2 min, and a final extension at 72°C for 7 min. The PCR products were analyzed by electrophoresis in 1% agarose gels, followed by ethidium bromide staining and photographed under UV light.
2.4. Construction of Recombinant Plasmids
The obtained PCR products were purified by using DNA gel Extraction Kit (Omega, Guangzhou, China) and cloned into pMD18-T (TaKaRa, Dalian, China) according to the manufacturer's instructions. Then the PMD18-T-g-α13 plasmid was transformed into JM109 competent cells. The positive colonies carrying the target gene were selected by fresh LB agar plates supplemented with 100 mg/mL of ampicillin after growing for 12 h at 37°C; then the PMD18-T-g-α13 was purified using the Plasmid Mini Kit (Omega, Guangzhou, China). The obtained plasmid and pET-28a (+) were digested with Bam HI/Hind III (TaKaRa, Dalian, China) and connected by T4 DNA ligase (TaKaRa, Dalian, China) to create recombinant plasmids which were confirmed by PCR and Bam HI/Hind III digestion and then sequenced by Sangon Biotech Company (Shanghai, China). The correct recombinant prokaryotic expression plasmids were named as pET-28a(+)-g-α13.
2.5. Expression of α-13 Giardin
The recombinant plasmids were transformed into E. coli BL21(DE3) (TransGen Biotech, Beijing, China). Freshly transformed bacteria were inoculated into LB medium (50 μg/mL kanamycin) and grew at 37°C with continuous shaking cultivation at 200 rpm until the absorbance at 600 nm reached 0.4~0.6. The α-13 giardin fusion protein was expressed under the induction of isopropyl-1-thio-β-D-galactopyranoside (IPTG); firstly, the expression conditions were optimized by adjusting IPTG concentrations (0.2, 0.4, 0.6, 0.8, 1.0, and 1.2 mM), induction time (1, 3, 5, and 7 h), and culturing temperature (16, 20, 25, 30, and 37°C). Then, they were expressed by autoinduction expression system (ZYM-5052); the bacterium solutions cultivated by ZYM-5052 for 3~16 h were collected, respectively. For details, refer to Studier [15] with few modifications. The cultures were harvested at 9600 ×g for 10 min at 4°C after washing the cell pellet in phosphate buffer (containing 0.1% Tween-20).
2.6. SDS-PAGE and Western Blotting Analysis
Samples containing 40 μL of total proteins were boiled for 10 min after adding 10 μL 5x SDS loading sample buffer, then stored for 5 min at −20°C, and centrifuged at 16200 ×g for 5 min. Then, 10 μL supernatant of each sample was analyzed by SDS-PAGE performed in Tris-glycine system, using 12% gels. Proteins were visualized by staining with Coomassie brilliant blue G-250. The target protein expression was detected by LiCor Odyssey infrared Imaging system (LiCor, Nebraska, USA) and analyzed by Quantity One 1-D Analysis Software (Bio-Rad, Hercules, USA). Ten microliters of each preprocessed protein sample was separated on a 12% SDS-PAGE at 120 V and transferred to nitrocellulose membrane (Roche, Indianapolis, USA) for 45 min at 120 mA. The membrane was blocked overnight at 4°C in 0.1% Tween-20-PBS containing 5% nonfat milk. The α-13 giardin proteins were detected using the ECL Plus Western blotting detection system (Tiannon, Shanghai, China) by rabbit anti-His tag monoclonal antibody (diluted 1 : 2000) and horseradish peroxidase- (HRP-) labeled goat anti-rabbit IgG antibody (Transgen, Beijing, China) at dilution of 1 : 3000.
2.7. Protein Purification
For purification of soluble α-13 giardins, BL21 (DE3) cells harboring pET-28a(+)-g-α13 were cultured in 1000 mL of LB medium for 5 h at 25°C with 50 μg/mL kanamycin. The cultivated cells were harvested by centrifugation at 8,000 ×g for 10 min, resuspended in PBS, and subjected to sonication (SCIENTZ, Ningbo, China). Soluble and insoluble fractions were separated by centrifugation at 10000 ×g for 30 min at 4°C. The supernatant was applied on a Ni2+ nitrilotriacetic acid column in binding buffer (0.3 M NaCl, 50 mM NaH2PO4, 10 mM Tris-base, and pH 8.0), eluted with different concentrations of imidazole (0.01, 0.02, 0.05, 0.1, 0.15, and 0.2 M), and analyzed by SDS-PAGE. The total protein concentrations in the supernatant were determined by using a Bicinchoninic Acid (BCA) Protein Assay Kit (ComWin Biotech, Beijing, China) according to the manufacturer's instructions.
2.8. Preparation of Anti-α13-Giardin Polyclonal Antibody
The anti-α13-giardin polyclonal antibody was prepared as described [16] with some modifications. Briefly, two male New Zealand white rabbits (2.5 kg) were inoculated with 1 mL (1 mg/mL) purified anti-α13-giardin protein emulsified with an equal amount of Freund's complete adjuvant via subcutaneous injection. Immunization of each rabbit was boosted four times by inoculation of 1 mL antigen mixed with an equal volume of Freund's incomplete adjuvant at 1-week intervals. Blood sample was collected 7 days after final injection; the sera were obtained to determine the antibody titer by indirect enzyme linked immunosorbent assay (ELISA). Prior to first immunization, a certain amount of serum was taken from ear marginal veins of each rabbit as negative control.
2.9. Determination of Antibody Titers by ELISA
The antibody titers were determined using indirect ELISA as described [17]. Briefly, purified α13-giardin proteins were coated on plates at 200 μL per well in 96-well plates at 4°C overnight. The plates were washed three times with PBS containing 0.1% Tween-20 (PBST). The coated wells were blocked for 1 h at 37°C with 200 μL of 5% nonfat milk in PBST, then washed three times with PBST, and incubated with 100 μL of anti-α13-giardin serum by serial dilutions (from 1 : 100 to 1 : 243,000, normal serum as negative control). After incubation for 1 h at 37°C, the wells were washed and incubated with 200 μL of HRP-conjugated goat anti-rabbit IgG antibody (dilution, 1 : 4,000; Transgen, Beijing, China). The reaction was terminated with the stop buffer, and the absorbance at 450 nm was measured using a microplate reader (Allsheng, Hangzhou, China).
2.10. Determination of Antisera Specificity
G. lamblia trophozoite lysates were extracted using Total Protein Extraction Kit (Vazyme, Nanjing, China) according to manufacturer's instructions. Total trophozoite extract and the purified α-13 giardin fusion protein were subjected to 12% SDS-PAGE and transferred to nitrocellulose membrane. The membrane was blocked at 37°C for 1 h with 5% nonfat dry milk in PBST, followed by incubation with the anti-α13 giardin antibody (1 : 200 dilution) at 4°C overnight. After three washes with PBST (containing 0.1% Tween-20), the membrane was incubated for 1 h at 37°C with HRP-conjugated goat anti-rabbit IgG (1 : 2000 dilution). Then the membrane was rinsed three times, and the protein bands were visualized with an enhanced chemiluminescence protocol and the Western blotting detection system (Tiannon, Shanghai, China).
2.11. Immunofluorescence (IF) Staining of G. lamblia Trophozoites
The immunofluorescence staining of G. lamblia trophozoites was carried out as described in previous reports [11, 18] with some modifications. Briefly, G. lamblia trophozoites were allowed to attach to sterile glass coverslips, fixed for 15 min with methanol, and permeabilized for 10 min at −20°C with 0.5%–1% Trion X-100/PBS. The cells were rinsed three times with PBS for 5 min each time. And they were incubated for 60 min with blocking buffer (5% goat serum) (ComWin Biotech, Beijing, China). Primary antibody was diluted in dilution buffer (at 1 : 500), after reacting with the rabbit anti-α13-giardin antiserum for 1 h; the samples were washed three times with PBS and then incubated for 1 h in the dark with Alexa Fluor 488 anti-rabbit IgG (H+L) from goat (1 : 200) (Beyotime, Jiangsu, China) at room temperature. Coverslips were kept in a humidified chamber to avoid evaporation. They were rinsed three times with PBS before counterstaining with DAPI (4′,6′-diamidino-2-phenylindole; Genecopoeia, Rockville, USA). After five washes with PBS, fluorescence staining was visualized by using Nikon DS-Qi2 Imaging fluorescence microscope (Nikon, Tokyo, Japan), with 100x oil immersion objectives (Size: 1608 × 1608), and images were processed using NIS Elements (version 4.50) Imaging software.
3. Results
3.1. Construction of Recombinant Plasmid
PCR amplification from α-13 giardin gene showed that the specific fragment was about 1000 bp, which was consistent to the expected size (Figure 1(a)). The identification of recombinant plasmid pET-28a(+)-g-α13 digested with Bam HI and Hind III was proved to be successful (Figure 1(b)).
3.2. The Expression of α-13 Giardin after Induction
The results showed that the target protein reached the maximum and accounted for 21.0% of the total cell proteins after induction with 0.6 mmol/L IPTG for 5 h at 25°C. Comparing the induction effect by ZYM-5052 with conventional IPTG induction at the same temperature, the results showed that the target protein reached the maximum and accounted for 28.8% of the total cell proteins after being induced by ZYM-5052 for 14 h at 37°C, while it accounted for 6.7% of the total cell proteins after being induced with 0.6 mmol/L IPTG for 5 h at 37°C (Figure 2(a)).
3.3. Identification and Purification of α-13 Giardin
SDS-PAGE results showed that both the supernatant and precipitate contained the target protein, but the supernatant had a higher amount of the target protein than the precipitate, indicating that the recombinant proteins mainly existed in soluble form (Figure 2(b)). The Western blotting analysis showed that the rabbit anti-His tag monoclonal antibody and HRP-labeled goat anti-rabbit IgG antibody reacted with a protein of about 40 kDa (Figure 2(b)), which is consistent with the theoretical value of the α-13 giardin. SDS-PAGE analysis indicated that the α-13 giardin fusion protein could be eluted with 0.1, 0.15, and 0.2 mM imidazole (Figure 3). However, the eluate with 0.1 mM imidazole contained several impurity proteins, while the one with 0.2 mM imidazole had loss of the target protein, so the eluate with 0.15 mM imidazole was more satisfactory. The protein detecting results revealed that the total protein concentration was 1.17 mg/mL.
3.4. Specificity of Anti-α13 Giardin Polyclonal Antibody
The anti-α13 giardin polyclonal antibody titer detected by ELISA was more than 1 : 243,000. The candidate antisera reacted with recombinant α-13 giardin expressed in E. coli as a polyhistidine fusion protein (Figure 4(a)) and were specific to a 40 kDa protein band with total trophozoite lysates in Western blot assay (Figure 4(b)). Results showed that the polyclonal antibody was specific to the α-13 giardin of trophozoites.
3.5. Intracellular Location of α-13 Giardin in Trophozoites
Immunofluorescence assay showed that the fluorescence signals of Alexa Fluor 488 anti-rabbit IgG (H+L) from goat were significantly distributed in the cytoplasm of trophozoite but not in the flagella and plasma membranes (Figures 5(a) and 5(b)). The preimmune serum (control) did not react with the trophozoite, and no fluorescence signal appeared (Figure 5(c)). It revealed that α-13 giardin was localized in the cytoplasm in trophozoites of G. lamblia assemblage A.
4. Discussion
Four classes of giardins, α, β, γ, and δ, have been identified as cytoskeleton components of the Giardia. The genome of G. lamblia possesses a total of 21 α-giardin encoding genes, which are α-giardins numbered from α-1 to α-19 giardin (α-7 giardin subdivided as α-7.1, α-7.2, and α-7.3). The different α-giardins have great differences in the protein expression level, the subcellular localization, and biological functions. The overexpression of some α-giardins often affects the division, proliferation, and differentiation of the cells, even leading to death of the parasite, which suggests that the α-giardins possess important biological functions.
This work presents the first report on prokaryotic expression of the α-13 giardin gene of G. lamblia. There are many factors that affect the expression of foreign genes, such as, the target encoding gene, the expression vector, the competent cell, the cultivation condition of the E. coli, and the induction methods [19]. Previous studies [6, 19, 20] have indicated that the genome of the eukaryotic protist Giardia is compact in structure, contains few introns, and has simplified machinery for DNA replication and transcription. And α-giardin genes have been known to lack introns, so α-13 giardin gene was cloned directly using genomic DNAs in the present study. The results showed that the amplification of α-13 giardin gene, construction of pET-28a(+)-g-α13, and the expression of α-13 giardin in BL21(DE3) were successful. However, in the initial stage of experiment, the production of target protein was very low and mainly existed in a form of inclusion body at 37°C as the optimal temperature for bacterial growth. Moreover, some expressing host cells had lost their expression plasmid. To overcome these limitations, we tried two solutions. Firstly, we compared the expression and solubility of fusion protein at 37°C, 30°C, 25°C, 20°C, and 16°C with different IPTG concentrations (0.2, 0.4, 0.6, 0.8, 1.0, and 1.2 mM) and different induction time (1, 3, 5, 7, 9, and 11 h); the optimized induction condition was for 5 h at 25°C with 0.5 mM IPTG to the recombinant BL21 at OD600 0.8. Secondly, they were expressed by autoinduction expression system (ZYM-5052); the results show that the quantity and stability of the functional recombinant plasmid in the autoinduction expression system had been improved. This work demonstrates the importance of selection of heterologous host and induction methods for successful production of soluble recombinant protein.
Different giardins have different subcellular localizations in G. lamblia trophozoites and may play different biological functions. Many α-giardins (α-2, α-4, α-5, α-8, α-9, α-10, α-11, α-14, α-17, and α-19) have been shown to localize in flagella; a few α-giardins such as α-3, α-5, and α-17 were found localized in the disc; some α-giardins such as α-1, α-2, α-7.2, α-7.3, α-8, and α-11 were located in the plasma membrane, and other α-giardins (α-6, α-7.3, α-12, α-15, and α-16) had spotty cytoplasmic distribution [6, 9–11, 13, 18]. Previous studies [6, 7] indicated that α-giardins are mainly involved in the cytoskeleton, flagella motility, membrane stability, and attachment of Giardia. Recent studies [21–23] revealed that α-1 giardin contains an epitope between amino acids 160 and 200 that is highly immunogenic and can stimulate the production of anti-Giardia antibodies (IgA and IgG2a), which is a suitable candidate antigen for a vaccine against giardiasis. In addition, compared with Giardia albendazole- (Abz-) sensitive clones, Abz-resistant clones had upregulated the α-2 giardin, which suggests that α-2 giardin may function as annexin to cope with reactive oxygen species in Abz-resistant clones [24]. These discoveries of the functions of α-giardins will aid in further research on the prevention and control of giardiasis. However, there are no reports about the localization and relevant biological functions of α-13 giardin to date. The present study on α-13 giardin localization extended the list of giardins located in the cytoplasm. Though the precise function of α-13 giardin remains unknown, its specific localization in the cytoplasm suggests that the protein may be associated with a particular cytoplasmic structure like free ribosomes or profiles of the endoplasmic reticulum and it may make contact with the cytoskeleton within the parasite. The exact subcellular location and specific biological function of α-13 giardin need further research in the future.
In conclusion, this work presents the first report on prokaryotic expression of the α-13 giardin gene and its subcellular location of G. lamblia assemblage A (strain WBC6). It may lay the foundation for further functional research on α-13 giardin of zoonotic G. lamblia assemblage A.
Competing Interests
The authors declare that they have no competing interests.
This work was funded by the National Natural Science Foundation of China (Grant nos. 31272551 and 31672541). G. lamblia trophozoite assemblages A were offered by Professor Zhaorong Lun from Southern China Research Center of Parasitic Biology, Sun Yat-Sen University, China. The authors would like to thank him and Dr. Yiting Xie for valuable contributions to this study.
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There are several technologies crucial to the Daily Deals space. Email delivery, Email management, eCommerce platform, analytics, scheduling, reward platform, social marketing, etc. Some of them should be looked at by Daily Deal companies in order to operate a more effective operation. Some future posts will mention several technologies while others will only focus on one. This post will focus on the popular technology of Online Appointment Scheduling.
In December of 2011, Groupon announced the acquisition of Canadian based OpenCal appointment scheduling software. (They bought them in September of 2011) Through Opencal, Groupon is providing a calendaring solution to merchants so they can provide better service to customers and improve the merchant's operational effectiveness. For merchants such as Spas and beauty salons, this is a necessity. Simply managing the scheduling of customers coming in from deal sites alone can be operational hell. An online solution geared towards helping merchants schedule better and perhaps improve customer retention and spend certainly adds value. It also makes sense to offer customers the ability of buying a daily deal and immediately book their appointment. This leads to better customer experience, less waiting times, better management of the schedule on the merchant's side and less returns from the customer.
So now that Groupon is offering such a service, the question is, "are other deal sites following suit?" Online scheduling solutions have become quite popular with merchants over the past couple of years and continue to grow in popularity. Who uses them? Spas, Beauty salons, Dentists, Orthodontists, Doctor Offices, Classes (yoga, dance, etc), Tutors, Pet groomers, etc. Are Daily Deal sites using them to help Merchants and their customers? More and more are. It just makes sense.
However, some of these systems are better than others, more expensive than others, some have APIs…most don't, some have the ability of Deal sites offering it to merchants as a value add, while others needs merchants to pay for it. So where does the evaluation start? There are hundreds of online scheduling software solutions at the moment. Which ones actually work within the Daily Deals space, which ones work in Canada, which ones can work in Quebec in French? Let us look at one leading online scheduling software that has been getting very positive attention in the Daily Deal market lately, Setster.
We have decided to focus on Setster because in April, they announced the launch of their API for Daily Deal companies, making it even more effective and valuable for Deal sites and Merchants to benefit from this type of solution.
Both Deal sites and Merchants find it challenging to properly handle the influx of customers from a Deals campaign. Poorly managed appointment scheduling leads to negative customer reactions and much higher rates of returns and loss of customers. None of which Deal sites or merchants can afford. Booking appointments immediately and easily after purchase is certainly one of the main areas to be improved in this industry. Yes, Groupon purchased their own online scheduling software company but most Daily Deal sites cannot afford the same strategy. Enter Setster's API; enabling Daily Deal companies to easily leverage the power of Online Scheduling for their merchants.
Setster's new API allows daily deal companies to integrate it's online scheduling solution both into their own website, as well as the websites of their merchants. Buyers of deal vouchers can, therefore, schedule an appointment with the merchant as soon as a deal has been purchased. This enables each merchant to allocate specific times and resources to deal with the demand of the deal campaign. As a result, daily deal companies are able to offer easy scheduling for participating merchants and ensure that every voucher purchased is redeemed, thus reducing returns and helping to convert consumers into longer-term clients for their merchants.
So, how can Deal services use Setster's API?
They can re-sell this service as their own through a white label option, or offer it to merchants for FREE, as an added-value incentive of working with them, or simply get the merchants to buy it themselves. Our last post, discussed how more and more Deal sites are looking at offering their merchants value-added technology in order to improve operational and marketing effectiveness. This is another example of that.
Here is what Edward Lujan, founder of Setster stated: "While daily deal sites and their promotional buyers are 100% web based, many neighborhood service providers have no web based infrastructure in place to deal with the influx of appointments, communication follow up, even up-selling and future marketing. We want to change this. The ability to transition the promotional buyer to the merchant is critical for the success of both the deal company and the merchant. Done wrong, the business will lose money on the promotion. Done right, the business will earn a happier long-term client: a client they can manage, market to, and provide other services to. It's about seeing a real, measurable return on their marketing investment."
We got authorization from Setster to promote their whitepaper on their API for Daily Deal companies. If you'd like to have a read, please get it here. If not, you can always grab it from Setster's API site.
It will be a good follow up in 6 months from now to see how many deal sites are leveraging the benefits of online scheduling for their merchants.


text: Digital Enfatico Lays Off 8% of Workforce By Joe Ciarallo|February 25, 2009 Share By Joe Ciarallo|February 25, 2009 Share Just several days after a video interview with PRWeek and DM News in which he stated, "We're in the throes of a lot of new business development," Enfatico CEO Torrence Boone announced the agency will decrease their workforce "by approximately 8 percent." From the agency's blog: Persistent cost pressures require that we reorganize our operations to better align our business with what is happening in the global economy. These changes also put us in closer alignment with Dell's new global structure and reduced marketing spend. We acknowledged these moves in an internal memo (see below to download) that we distributed today. AgencySpy is reporting that the cuts equal about 64 people out of Enfactico's 800 strong global workforce. PRWeek's Aarti Shah says the agency currently employs "30 full-time staff on its PR team, which makes up less than 4%" of total staff. No word if PR was among the cuts. Share http://adweek.it/2l0lEpU copy Adweek Adweek


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Back in September I was contacted by the Innovo team to attend the launch of their new product at the lovely Soho Hotel in London.
The event marked the launch of a new campaign which is all around Inner Strength, in particular, the strength of the pelvic floor! I know. I can see you rolling your eyes and hiding away… but please don't.
It's such a shame we tend to discuss everything else but when it comes to our pelvic floor, we don't or are ashamed of doing so. That's why I was even more interested in attending and testing this product, and let you know my thoughts on it.
Pelvic Floor
Pelvic floor weakness affects an estimated 1 in 3 women in the UK, particularly following childbirth, yet many cases of stress urinary incontinence cases can be reduced or eliminated by pelvic floor muscle training.
There are also other benefits to a strong pelvic floor including increased sexual pleasure! Ehm, ehm, ehm. Now I get your attention!
Pelvic floor weakness affects an estimated 1 in 3 women in the UK & 1 in 10 men!
The evening started with Jane Wake, a pelvic floor Pilates instructor, telling us all about the importance of exercising our pelvic floor. She also explained where we should find our muscles and what exercises we can do to maintain our healthy pelvic floor.
Then the INNOVO team presented the newly launched unisex shorts and explained all the features and benefits.
INNOVO uses a patented EMS system to provide a convenient, garment-based solution treating the underlying causes to help women build their inner strength.
The evening was wrapped up with a performance of a stand-up comedian who is back from the Edinburgh Fringe where her show 'Laugh, don't leak' received 5* reviews and no wonder. I hadn't laughed that much for a long time! That was a good test to see how strong my pelvic floor is!
Why INNOVO?
– non-invasively strengthens your pelvic floor
– clinically proven, a long lasting solution to treat the root cause of bladder weakness
– suitable for both men and women
Bladder weakness, urinary incontinence, little accidents, whatever you call it, this is what commonly happens when we cough, sneeze, laugh, or feel a sudden urge and have to go straight away.
A weakened pelvic floor can be caused by pregnancy, surgery, heavy lifting, pelvic/prostate surgery, menopause, high-intensity workout and being overweight. 1 in 3 women and 1 on 10 men suffer from leaks every day!
… But because the pelvic floor is a muscle, you can strenghten it!
As a fitness blogger behind Fitness 4 Mamas, and a mother of two, I know how important it is to exercise my pelvic floor.
On the other hand, I am sad to say I don't do so as much as I should. At the end of the day, every muscle needs to be exercised to be strong and working properly, so pelvic floor should be no exception. It all makes sense, doesn't it?
As a keen runner, I had a few "accidents" while back to running soon after giving birth to my second daughter. Luckily it's improved, but when my girls ask me to join them on the trampoline, I am still reluctant as I feel like it might happen again… Yes, it happens to the best of us and there is really no shame to talk about it!
Soon after the product launch, I was sent a pair of shorts to try. I went for the small size (UK10-12). Then I carefully read the product manual.
I highly recommend this as there are loads of tips of what programme to choose, how to wear the shorts correctly, and how to solve a few issues.
Then I put my shorts on and started getting used to the intensity of the impulses. It took me a few days to reach the recommended intensity for my size, and also to find out which position I was most comfortable to be in while wearing the shorts.
It has been 6 weeks since I have been using shorts. I am so pleased to say I can actually feel how my pelvic muscles are getting stronger.
Before using Innovo shorts I struggled to squeeze and exercise these muscles but this is changing, and I am not stopping. My aim is for my pelvic floor to be so strong to make me confident enough to join my two daughters on the trampoline!
Another great thing about this is that they see me wearing the INNOVO shorts and know why I am doing so. The educating about the importance of pelvic floor has started early in our family, which I believe is a great thing. What do you say?
Featured post


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looking at meraki, you see an RV with its universal patented look: curvy streaks across the body, maroon and gray with the Dalmatian dog panting above "Coachmen Freelander." The ass of mer sticks longer from the rear tires, giving you indication that it's bigger than 26 feet long. The only sign of character you would see is "yes, I am the cat lady," magnet stuck on the cover case of where propane releases when using the heater.
That pretty much reflects the outer part of who we are. If I had it my way, there wouldn't be a sticker of some curvy sorts. Fuck conformity. There wouldn't be that out of breath Dalmatian hovering over the word "Coachmen" (sexist much?). The only thing I am okay with is the term, "Freelander." Otherwise, it's just a manufactured crap telling us its sticker price's worth. She's far more storytelling than that uniformed look.
She's our home and inside that manufactured body, she holds hundreds, if not, thousands of memories that will last forever.
Now, as I sit outside with my coffee and enjoy this gentle Texan breeze, I just thought of an idea. I want to find a decal to add a cape on that dog (that breed is known to be Deaf, did you know that?), with a tiara on its head and to add two letters "w-o" in between the h and m. Then whoever notices those slight creative adjustments would definitely get insight of who I am [we are].
the heart of the deep south
As a liberal Californian mother of three boys who have Jewishness in their blood, going into the Deep South had some reservations from me. Especially when the majority of them voted for Trump. Not just voted, they believed in him. When doing so, they believed in so many things that basically decry on the very humane aspect of love and humanity. When one goes against it, how is it possible for them to have a speck of iota of spirit in kindness (when they are extremely conditional and selective)?
Needless to say, my judgment and reservations toward the people of the Deep South is harsh, filled with bitterness and resentment… and disbelief. Trumpets, I call them. When I call someone a "Trumpet," consider it the deepest level of offence I can offer, with such disdain. To this day, I still hold them accountable for the hatred we have in our society, especially to the people of color, to the people who want to love equally and ultimately, personally close to my heart, to the women.
We were knee deep into the south the past few months. I've come to a conclusion. It's with profound sincerity when I say this, the Deep South has the heart of the country, but it's so flawed. They were the friendliest, sweetest, kindest and charming bunch. They were patient, they were helpful, they handled our Deafness far superior than those people in the blue states. It shocked me to the core. Everywhere I went, be it a drive thrus or at a general store in the middle of nowhere, they all somehow knew sign language. They all called us "honey" or "sweetie." They did not make me feel inferior, but rather treated us as a revered novelty. They welcomed us as if we were family members and offered us kindness even if we were complete strangers.
But, how in the world can they be as heartless as my label, "Trumpet"?
The other day when Oisin made a new friend with a complete stranger, I sat and watched them play and chat together. The boy's mom was also watching. We waved at each other and I went over to her and we started talking. It was beautiful, us getting to know each other, and my son getting to know hers (they were talking about the book titled, "wrinkle in time."). She asked me if Oisin's name was Irish and I almost started telling her that my kids' names all had both ancient Gaelic and Hebrew names because they are of Jewish descendant. I hesitated and decided to jut say, "yes, we chose ancient gaelic names."
She smiled and looked at our boys, the sun was descending, it was super warm and you could see both boys laughing as they were trying to communicate.
It was beautiful, but my fears in telling her that my boys are also Jewish disturbed me. I don't think that the mother realized that she made me feel uncomfortable in the fact that I would have potentially endangered my boys if I told them that they were not just Irish, but also Jewish. To be fair, she probably is a kindred soul and not deserving of my assumption.
That's how it is here, for me, in the Deep South. As sweet as their iced tea are, they coated sugar over their self-serving steeped beliefs into what makes America Great. Being sweet and all, but totally going dark on what really matters and that is, being a humane human.
good-bye south (warmth)
One year isn't even enough. With such a limited time frame, we are pressed to move upwards, into the cold zone. I am trying my best to be appreciative of the fact that we are living as a full-time family, but honestly, the weather was such a crappy friend on most of our trip. I mean, we had deep freeze for most of our stay at our beach resort (this never happens they claimed), then we had the wacky cold weather that started snowing out of no where when we were in DC (that is such a weird weather!! they said)... oh and the south carolina farm? "It's been raining and so cold, which is unusual for us." *eye rolls* Apparently, the cold front pretty much followed us throughout the entire trip.
And now? We are following it because we have to go north. If you hear a frustrated growl emitting from a lady, it's from me.
Well then, if this trip taught time one thing, it's that planning ahead can be a bad idea and not planning well pretty much has the same result. I know that seems confusing, trust me, it's the truest way I can describe the rv-planning. Best thing we can do is to shrug our shoulders and try and make the best out of it.
We headed into eastern parts of Tennessee, fully aware of 20's-30's temps for the week. I've pretty much given up on having any hopes of us sitting outside of Meraki and eating our meals at a picnic table with an idyllic weather.
I knew it'd be cold and that there was a possibility of snow at Crossville, Tennessee. What I did not expect was the beautiful snow that graced us this morning. I was thrilled when I saw thick snowflakes floating down and everything looked so clean and peaceful, so excited that I woke my boys up (Eitan, being a tween he is, went right back to sleep). I am glad I did because it was short-lived. Literally, within an hour the snow was gone.
Then new problems came up.
My boys noticed water dripping from the window frame in my bedroom. We never had that issue with all THOSE rain and cold weather we've had lately, today was the first time it happened. I realized that the snow on top of Mer melted and pooled into one specific area, which obviously got right into a crack in the window frame. I went up on the roof and put a towel there, a very redneck DIY approach, and the leaking stopped immediately.
I thought to myself, it's official, it'll always be a "Today's weather is a 100% chance of Be Shitty to Meraki."
It looks like I gotta take out the window frame (Fulltime Families on Facebook said it's easy to do) to dry it, putty it and caulk it, but I cannot do it because I need someone else to "catch" the window as I pop it out. Errrrmmm. The "resort" we are at (laughing so hard) is in the middle of nowhere and we're the only ones there. Guess I gotta wait until the next resort, as it's much bigger and sorta fancy. I'm sure there will be someone who will help. This will add another notch in my belt of being a handy RV Lady.
Anyway, I bid thee farewell to the south, to any possibilities of warmth, thanks for a fabulous time. And now, onwards to our other half of the journey up into north pole.
six months
And so, just like that, half of the year just arrived. Are we wiser? Travel worn? Wanting more experience? Ready to go home? Not ready to go home? We're all of that.
A dear friend of mine asked me a question in a way that nobody else ever did. I mean, people have asked about why I am doing this trip, how did I think of this trip, etc etc etc. But when he asked me with an emphasis of "you" as what did I REALLY want out of this?
I found myself rambling an answer and finally replied, "me. I want myself, me out of this."
And so far, I think I am succeeding with that.
just a mother who fled from society's constraints and is super excited to wake up to the outdoors, remain braless daily and teach her boys the art of boredom and discovery.
September 2018


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The Brunel University Prize for African Poetry
The brunel University prize for African poetry seeks out the best poems from poets across Africa each poet submits 10 poems, the shortlist was released on the 16th of march 2017, and the winner(s) will be announced on the 2nd of may 2017
Sahro Ali
Her Nationality and Poem
Sahro is from Somalia
In Morocco two girls are jailed for homosexuality.
My mother pulls apart my legs in fear with her
Filching fingers.
Where is my daughter? Is she here?
Mouth shut with hot wax, burbling and yellow
A bee's nest has found itself in her throat & she tries
To cough it into me.
You are my daughter are you not?
Her friend tells her the best way to keep me safe is
To sew me shut, 'a bird cannot fly if you clip its wings.'
My mother's fingers shake around the needle and thread
She's a quiet, polite madness, her cheeks fat with salve
I'm just trying to find my daughter I'm sorry
There's something crawling down my stomach but it can't
Find its way out, I stick my hand up my pussy one last time.
I birth a black dog, and let my mother lick the meat off its bones
A bee pops out onto the surgery table and I try to speak through
The honey in my mouth.
I put my hand on top of her shaking ones, teeth at the thread
Its fine you've found her
Leila Chatti
Her Nationality and Poem
Leila Chatti is from Tunisia
fasting in tunis
Longing, we say, because desire is full
of endless distances.
– Robert Hass
My God taught me hunger
is a gift, it sweetens
the meal. All day, I have gone without
because I know at the end I will
eat and be satisfied. In this way,
my desire is bearable.
I endure this day
as I have endured years of days
without the whole of your affection.
Your desire is one capable of rest.
Mine keeps its eyes open, stalks
through heat that quivers,
waits to be fed.
The sun burns a hole through
the sky and I am patient.
The ocean eats and eats
at the sand and still hungers.
I watch its wide blue tongue, knowing
you are on the other side.
What is greater: the distance between
these bodies, or their need?
Noon gapes, a vacant maw—
there is long to go
until the moon is served, white as a plate.
You are far and still sleeping;
the morning has not yet slunk into your bed,
its dreams so vast and solitary.
Once, long ago,
I touched you,
and I will touch you again—
your mouth a song
I remember, your mouth
a sugar I drink.
Kayo Chingonyi
His Nationality and Poem
Kayo is from Gambia
The Colour of James Browns's Scream
for Stephen McCarthy and Todd Bracey
I have known you by many names
but today, you are Larry Levan,
your hand on the platter, in the smoky
room of a Garage regular's memory.
You are keeping When Dove's Cry
in time, as you swing your hips,
and sweat drips from your hair
the colour of James Brown's scream.
King of King Street, we are still moving
to the same sound, though some
of us don't know it is your grave
we dance on, cutting shapes
machismo lost to the beat–
every road man is a sweetboy
if the DJ plays Heartbroken
at just the right time for these jaded feet.
Teach us to shape-shift, Legba,
you must know I'd know your customary
shuffle, that phantom limp, anywhere;
that I see your hand in the abandon
of a couple, middle of the floor,
sliding quick and slick as a skin-fade
by the hand of a Puerto Rican clipper-man
who wields a cutthroat like a paintbrush.
Let us become like them, an ode
to night, ordering beer in a corporeal
language from a barman who replies
by sweeping his arms in an arc,
Willi Ninja style, to fix a drink our lips
will yearn for, a taste we've been
trying to recreate ever since.
Saddiq Dzukogi
His Nationality and Poem
Saddiq is from Nigeria
father's demise
the opaque face of things
like stone & water
& my extended family fighting
while the village expects us to soak
in a seawater of mourning
father's demise is a dispersing light
I kept grumbling at the moment
of his passing my siblings each
are trying to hide their happiness
my shadow is only good at imitating
my posture only grandmother owned
a genuine grief the moon hanging
by the window is unable to wash off her sadness
night won't penetrate her eyes
my father's brother looks like he is hiding
his schemes intimate like a lonely wife & her pillow
he has always held what is father's
in the same way a best man
looks at the bride he is secretly in love with
my mother once told her friend
he had come to her tiptoeing
wanting to wear my father's shoes
but later found my mother's body
a room too big for his foot
my mother didn't know I broke the meaning
of her metaphor like she breaks kola
for those who have now come to mourn
sad stories are stretching the size
of our sitting room into a market
I locked myself in a dirty toilet
the stench there less
than a family's hypocricy
Yalie Kamara
Her Nationality and Poem
Kalie is from Sierra Leone
Mother's Rules
For my mother
I. If you see me praying in the living room, never sit in front of me. You are not God.
II. When we go to a restaurant and I don't know any foods on the menu, never order me a meal that is spelled with silent letters. I came to eat, not to explore
III. You didn't make food. No. God, did. You cooked food. Watch your English. Watch your faith.
IV. Your Krio is offensive. When you speak, you sound like Shabba Ranks. Your accent is funny, but keep practicing. It is the only way we will be able to gossip in peace while at the supermarket.
V. Try to learn the language of your lover and his family. They could be smiling to your face and getting ready to trade you for 6 goats and 3 mules during your first trip to their homeland.
VI. If anyone stares at you for too long (more than 5 seconds), start speaking an imaginary language while maintaining eye contact. They will be the first to look away.
VII. Consider the consequence of purchasing human hair wigs, second hand clothing, and used furniture. Maybe you will feel beautiful, and also save money, but you never know whose bad luck or misfortune will be sitting on your head, body, or in the home in which you sleep. Buy what you can truly afford.
VIII. Your father's Muslim, so you are too (1989-1993).
I am Christian, so you are too (1993-2012).
I am Catholic now, but you keep praying (2012-present).
IX. You laugh at me now. Like I laughed at my mother. Like she laughed at hers. Like your daughters will laugh at you. And I will live long enough to forgive your folly.
X. Just make sure to pray.
Kechi Nomu
Her Nationality and Poem
Kechi is from Nigeria
Note to the Boy Kicking the Stone
The boy's body
to pick the stone
And you see how
his body too is a road
with curves,
too many
Once or twice,
the bones
of his
Here, a story begins
rises, falls
and ends
Richard Oduor Oduku
His Nationality and Poem
Richard is from Kenya
The Old White Man and the Bungalow on 5th Avenue
He walks the pavements
in boxer shorts
too brief for a stuffed groin
and spindly legs
that drop flakes of dry skin
with each laboured step
he never leaves his fez behind
his browline glasses
nor his book, the Tibetan Book of the Dead
but you can tell he is most happy
on Saturdays when he stops
to speak to streets boys
and give them sweets and bread
we are seated on the terrace, nursing whisky
and Shangri-Las' Out in the Streets –
a mix of teen-beat puppy love and nostalgia
sitar drones and psychedelics:
a world of Beatles and porous borders
then he leans forward and tells me a story
"we started from the Atlantic shores
on a forlorn stretch of brick-red outposts
and centuries-old souks
singing on the backs of camels
battling winds and scorpions
and the silence of the Sahara
in the kingdom of sand
the only god is an oasis
and when the pink skies poured over us
we cuddled in desert camps
and emptied our tote bags of want
into each other – under pickled stars
everything was fine, everything!
until that summer of 1984 in Morocco
when I was stirred from the milkshake
of sleep by the hiss of a horned viper
slithering away from a cold body:
the guile of the desert took her away
I left her bones to desiccate in the sands
and escaped the haunt of shared memories
to Kenya – a country I barely knew
to start a new life of abridged joy
in this bungalow on 5th Avenue."
Romeo Oriogun
His Nationality and Poem
Romeo is from Nigeria
Invisible Man
And the voice was a lost bird embedded in a boy
like a word stranded between pages.
He said flee from the heat wrecking your body
and you ran to a place where water
running over pebbles is a whisper of wildness,
where lost boys are birds hiding their heads
under wings as they touch their wetness
in the dark and whisper hallelujah.
The radio said, a father shot his son for loving another man.
Marvin Gaye lives in the heart of a black drag queen
and to be a song of pebbles and water is to run into a city of light
and surrender your throat to the song of a bird.
On the streets of Lagos, a boy searches for himself in mirrors,
he opens his heart and hears the voice of his father
breaking his bones into a prince
collecting burnt teeth lying as warning on holy grounds.
This is how we kill love;
hunting it in the dark when it is soft
like a newborn chick,
breaking its bones till it becomes
a boy filled with dead men.
Rainfall teaches the ground how to breed:
a boy learns about the wetness of his thighs on a cold night.
Poster of boys diving into water holds him in a trance.
A horse hears the coming of speed rising in his blood;
a horse responds to the call of wild hills as water tickles the sky.
Wet dreams:
a boy hears the whisper of another boy deep in his bones
and wonders about the origin of stars,
his body is a lamp learning how to give light
in a place where a boy opens his mouth
from the door of a tomb;
where a boy takes his first breath
and resurrects into life;
where a boy learns how to make honey
out of a skin.
This is how to live:
a resurrected boy hides in dark bars and stare at muscles of hard men.
He is called Joe, he is called John, he is called The Wind
and that is how to be unseen.
And this is real:
a man hides his voice in a throat
before bursting out into songs.
Verbs are boys learning how to kiss,
like you turning your body into a blue sea;
turning your lips into pictures of love.
Like you opening your body into a little island;
opening your skin into a beautiful world.
Verbs are boys learning how to love
in a place where death lives in water.
One step at a time. A boy learns how to dance,
his voice is a stream learning the music of the ocean.
He opens his mouth and paints blue skies with the magic of flying.
He opens his hands and flowers plait the air with music.
One step at a time. A man kisses another man
and hears bullets hitting his windows.
A man kisses another man and hears a mob running on his skin.
A man lies on the edge of bliss and hears the rape of boots on doors,
still we rise with the sun and plant seeds of love in dark places;
still we love and hide and wait for rapture inside a boy's body
as a voice flirts with the birds in his throat,
while a man burns on a street in Lagos for singing too loud
Rasaq Malik
His Nationality and Poem
Rasaq is from Nigeria
After My Grandma's Burial
We sit on the benches at the
facade of our house, my mother
in a dark gown, my father attending
to visitors, the children staring at the
grave of my grandma, their thoughts
too frail to decipher the meaning
of burying a beloved. At night we
remember those who are not lucky
enough to have a decent burial,
those who lie in unknown places,
those who find solace in caressing
the photographs of their lost relatives,
those who are murdered in the cold
of war, those who wake up to see
bullet holes on their doors,
those who learn how to pray as
they wilt in the fire of bombs,
those who pass through the dark
as they carry the burden of war.
At night we listen to the voices
of our beloveds in the walls
of empty rooms, in the leaves
that sway as wind blows, in the
silence that thickens as we
remember how death happens,
how we become dust, scraps in
the kitchen, fossils for earth to
devour, bones that never rise
no matter the force of rain.
Nick Makoha
His Nationality and Poem
Nick is from Uganda
Candidate A
For the record, he loves his own reflection,
this farmer's son from the delta. A splendid type.
Bone from the neck up, trained in wickedness,
born to lead, useful against the Mau Mau.
Unprovoked, he once cracked a cow's skull
with flat palms as the beast stared at him.
Did the same to three cattle herders at Lake Turkana.
Reached into their necks to eat their intestines (allegedly).
He should have been court-martialled,
especially after the assassination attempt.
Such men rise in the ranks and can only be removed
by death or revolt. I suggest we seduce him with wives.
Surround him with ceremony, regulation and rules
even though secretly he feels they do not apply to him.
Feed him with titles: His Excellency, Field Marshall,
Effendi etcetera. His cravings have no limit.
We can use distrust of competition to our advantage.
He will demand acclaim in an unbridled urge to destroy.
Easily compensated, he does not accumulate goods
or possessions for the future, opting for immediate gratification.
While he buys friends, kills citizens without fear of god or religion,
in his effort to be remembered, we will make our mark.
Previous winners of the prize
Warsan Shire
Liyou Libsekal
Safia Elhillo & Nick Makoha
Sudan & Uganda
Gbenga Adesina & Chekwube O. Danladi
Both Nigerians
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She always carries a notebook with her as ideas come to her at the most inopportune moments, once Lisa has one idea then another couple usually stem from that one and she have been known to run back and forward to her craft space while making dinner!
Lisa loves sitting down to develop the ideas, they don't always work out but when something does, she says it's an amazing feeling, especially when someone either comments on it or buys it.
She believes in the handmade and all the love and quality that go into making a piece from scratch. Lisa believes in having beautiful things that are unique and a little different.
Lisa designs and makes all of her products using resources around her. She uses vintage threads, some of which are older than her, passed down from various grandparents and aunts! She upcycles books from local charity shops for her jewellery pieces. She also uses vintage buttons and fabric she have accumulated over the years including Liberty fabric and Lisa buys her Harris Tweed directly from the weaver. Other materials are sourced from mainly Scottish independent companies.
Lisa can be found on Facebook, Twitter and Instagram under @handmademoment5
You can visit her website.


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Add Trim To Round Throw Pillows
Round throw pillows – A well-chosen throwing pillow can instantly transform the dullest looking sofa or armchair into something sophisticated and eye catching. Similarly, a uniform pillow can easily make the exact same furniture even more common. Fortunately, when it will throw pillows, it does not take much to change them. For example, some well-placed trim will immediately make your throwing pillow appear more elegant and exotic. Using trim also means that you do not need to buy new pillows.
When a big round throw pillows with burlap, seat cushion or pillow cushion is just too much bag worm for the bedroom, a boudoir pillow is a modest option. Sack weave collects and tuck lightly, but seems to miscast in voyager and sews down for folds that hold. Boudoir cushions can be small cylinders and squares, narrow rectangles, chubby triangles or simple round discs. The fabric of the fabric, its antique, unbleached and pale, or faux pale, stamp or design should be the center of attention. Keep the pillow design simple, use seam or button close to detail. And throw the pillow in a pile of other rustic textures, matte finish and vegetable-colored colors.
How to add trim to round throw pillows. Measure the perimeter of the pillow. Write down these measurements and cut a piece of trim that is long enough to stretch around the entire perimeter of the pillow, plus an inch. Line your trim up with the seams along the perimeter of the pillow, needle trim each 2 inches. For tasseled trim, rip the seams off the pillow open, one side at a time, and put the top edge of the fringe in between the sides of the fabric, for a fancier look if you want. Whichever option you choose, pick up the trim in place.
Then to add trim to round throw pillows. Check the trim to make sure it is straight. Adjust if necessary. Sew trim directly on the pillow using a sewing machine; remove the staples as you go. If your pillow is too uncomfortable to sew on a sewing machine, sew trim on. And then make small, narrow, almost invisible stitches in a thread that perfectly match the attire. Tips and warnings. You can add extra decorative items when you finish installing trim. For example, add items like buttons, rosettes or beads to the edges or fronts the pillow, and also sew them all by hand.
This gallery is about round throw pillows.
This post topic: Interior


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Photo Album | YouTube | Professional
Theatre Shakespeare
Nathan and Tanya have been season ticket holders at both the Actor's Theatre of Charlotte (ATC) and at Atlanta's Shakespeare Tavern. ATC our favorite place in Charlotte and the Tavern is our favorite overall theatre in the world (even better than The Globe Theatre in London). The Shakespeare Tavern is great, it is not overdone, the players are down-to-earth, the sets are creative, and the atmosphere is a ton of fun.
VA Tech Football
Our entire family is a big fan of Virginia Tech football and fans since 1999. We try to make it to as many games as possible including away games.


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A note from the field
Oct 21, 2018
Autumn and Knitwear Essentials for November's No1 Event:
Every year around this time, the team at Smart Turnout relish in the knowledge that the 5th of November is fast approaching. The days draw in and the temperature falls. Autumn is at last underway, and the smell of oaky bonfires graces our nostrils. With a biting chill in the air it is certainly time that the Gentleman learns to embrace the cold and invest in some knitwear essentials to see him through the crisper months. Too often, style is lost in the bid to remain snug and that, our friends, simply won't do!
Remember Why…
Bonfire Night is a true indication of Great British spirit. Turning its morbid history as one of the most treacherous plots in UK history, in to a celebration of Britain and its ability to prevail in even the bleakest hour. In 1605, Guy Fawkes himself was famously arrested for conspiring to blow up the House of Lords. Consequently, since then, Britons have gathered annually to burn effigies of Fawkes in celebration of the plot's unsuccessful end. Over the years, the skies on the 5th of November have been illuminated by the wonderful multicoloured hues of dazzling fireworks and as a result the evening has become somewhat of a social occasion. The night is enjoyed by families and friends, who gather across all corners of the UK. Amongst the Catherine Wheels, Rockets and Fountains, it has become increasingly difficult to truly steal the spotlight.
Remember How…
The British gentleman, as ever, should come to Bonfire Night prepared. What with Bonfire Night being of such historical significance, is an opportunity to come dressed to the nines.
Looking to stand out amongst the bustling crowds? Longing for socks that will complement the bonfire? Frustrated at neighbours admiring the fireworks rather than your matching hat and scarf? Then you have come to the right place. Below are some go-to men's knitwear essentials , that you should endeavour to remember, remember, this 5th of November:
All of our British Cricket Sweaters are made to order and handmade in England from pure wool. This darker, navy sweater is the perfect thickness for facing the autumnal night, promising to keep you warm without compromising your undoubtedly dapper outfit. The sweaters themselves belong to a tradition just as old as Bonfire Night and are one of our knitwear essentials. A timeless piece for any British gentleman's wardrobe.
Our sweaters come in a range of Autumnal colourways and available to purchase on our site.
This double thickness, hand pressed, Saxony wool scarf will certainly attract admirers. Enviably cosy, this scarf has been made in the Harvard University colours. The dark burgundy will be right at home amongst the autumn leaves and the amber flames, while the white provides a crisp accent. Furthermore, if you are lucky enough to have a warm evening on the 5th, this scarf will look just as good hanging loose over your shoulders.
Understanding the importance of even the smallest details, we at Smart Turnout believe that perfect socks are critical. The Argyll & Highlanders socks, made in a high-quality cotton poly mix, provide the ultimate comfort whilst continuing the traditional autumnal theme of your accessories. Above all, the regiment's colours – navy, yellow, dark green, and red – should match everything from the field below to the fireworks above.
Similarly, the Royal Navy equivalent, made up of thicker, accented stripes of navy, red, and white, epitomise British tradition. Whichever you choose, you are certain to outshine even the burning Fawkes. Feel free to follow our lead and check out our other autumnal favourites for essential men's fashion.
Make this November 5th one to Remember.
We hope you have acquired some new ideas on which knitwear essentials are crucial for the season. Finally, please comment below what you would wear to smarten up your Bonfire Night, we love to hear your own personal styling tips.


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Sorry for the delay, Gentle Readers. An inattentive blogger who will remain nameless neglected to properly schedule the post.
Part One
G.P. Lowery and C.W. Babcock exchanged words with the official printer for the Kansas legislature, John H. Brady. He told them bluntly that Andrew Reeder authored all Kansas' sorrows "and they must have his head, if they had to go to Pennsylvania for it." If the free state embassy let their danger in the presence of bands of men armed, hostile, and likely lubricated, slip their minds for a moment, this surely recalled the peril. Lowery told the Howard Committee that things began to get more heated, so he and Babcock resumed their trip to see Wilson Shannon and deliver Lawrence's letter. Along the way they saw more and more armed Missourians, including a fellow so far in his cups that he had a cornbread breakfast in one hand and a wagon wheel in the other. Duty drove them on and thus whatever story got this man to that point passed unrecorded.
The free staters arrived at Shawnee Mission shortly after sunrise. They, unlike Franklin Coleman's party, found Wilson Shannon present. He took their letter and read it through:
I do not know whether that letter is anywhere in existence now. I wrote the letter, and it was signed by Governor Robinson, Colonel Lane, Mr. Deitzler, myself, and four or five others. The contents were, that he might not be aware that there was a large mob collected on the Wakarusa, who were stopping travellers and goods, and plundering the country; and that we took that means of informing him that was the fact, and that they claimed to be there by his requisition; that we wished to know if that was the fact, that they were there by his authority; and, if so, whether he would remove them, and prevent these depredations, or compel us to do it ourselves, by resorting to other means or higher authority.
If the letter survived, I haven't found it. It doesn't appear in the Executive Minutes where it ought to, nor does Robinson give a copy despite referring to Lowery and Babcock's mission. Whatever happened to the original thereafter, Shannon read it and promised a response. After a while, he called the envoys back and they discussed matters:
He said there had been sixteen houses burned here by free-State men, and women and children driven out of doors. We told him we were sorry that he had not taken pains to inquire into the truth of the matter before he had brought this large force into the country, which, perhaps, he could not get out again; and that his information was wholly and entirely false, as nothing of the kind had happened.
At the time, one imagines that Lowery and Babcock found a more diplomatic way to put that. The free state party often blamed the arsons on persons unknown, or on proslavery men trying to frame them, but they can't have fooled many. Rumors do fly, but clearly Charles Dow knew who fired the cabin on the claim he jumped. The accusation appears in proslavery sources often enough, and with consistent enough numbers, to argue that antislavery Kansans had taken up the habit. Those displaced households, Coleman's included, came from somewhere. If Lowery and Babcock, who hardly lived at the far end of Kansas from Hickory Point, didn't know the fact then they must have worked hard to stay ignorant.
Shannon then, per Lowery, denied knowing anything about Missourians coming into Kansas to work their vengeance on Lawrence. He might not have at this point, but that seems like a stretch given about the time he met with the Lawrence envoys the governor also asked for federal troops to manage the situation. Either way, he held the free state proclamations about the laws of Kansas against them. From the context, it seems Shannon justified his summons of the militia on those grounds.
We explained to him that the Territorial laws had nothing to do with this case; that we were getting ready at Lawrence to fight for our lives, and the only question was, whether he would be perticeps criminis to our murder, or the murder of somebody else, should we all be slaughtered. We explained to him, that the rescue upon which he based his proclamation took place a number of miles from Lawrence; that there were but three persons living in Lawrence who were alleged to have had anything to do with it, and that they had left the town, and were not there at all from what we could judge of the intentions of the force at Wakarusa, at Lecompton, and in the county about, from their own declarations, they intended to destroy the town for a thing in which they had no part or parcel.
Lowery might have lied to Shannon's face, but not about that. The proslavery army, to judge from the words and conduct of its leaders, aimed to rid itself of Lawrence once and for all.


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Central Arizona Landscape Management has been serving the valley since 1979. We specialize in Landscape Design (also known fondly as 'exterior design'), Commercial and Residential Installation, and Commercial Maintenance


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upcoming events
March 2015
tcworld India conference 2015: Putting the spotlight on Indian technical writers and translators
The tcworld India conference 2015 has continued its successful path with the 5th international conference held in Bangalore from March 1213, 2015. More than 300 participants took the opportunity to learn from an impressive line-up of internationally renowned speakers.read more...
November 2014
Visitor record at tcworld conference 2014
From November 11-13, the tcworld conference 2014 and tekom fair welcomed about 4100 participants and fair visitors from all over the world at the new venue at the ICS International Congress Center in Stuttgart. It thus confirms its position as the leading global event and marketplace for technical communication.read more...
October 2014
Proven concept, new location: tcworld conference moves to Stuttgart
For the first time since its launch in 2005, the tcworld conference will be held in the International Congress Center in Stuttgart. The state capital of Baden-Württemberg is renowned for its fast cars, high-quality living and its hard-working, down-to-earth locals.read more...


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Pablo Picasso Art Museum in Münster
The only Picasso museum in Germany exhibits the artist's lithographic works
Germany's only Picasso museum is exhaustive: it contains nearly the entire lithographic portfolio of the world-famous Spanish artist.
Despite its young age, the Pablo Picasso Art Museum in Münster, which opened to the public in 2000, has proved to be a tourist magnet for Picasso fans from around the world. The collection provides comprehensive insights into the lithographic works of the artist – in fact, it houses nearly the entire lithographic oeuvre of the world-famous Spanish artist. It also includes state prints and test prints, which give visitors a clear picture of Picasso's artistic production processes.
Germany's first and only Picasso museum uses its 600 square metres of exhibition space to display regularly changing presentations covering Picasso, his contemporaries and his artist friends. By continuously examining different aspects and excerpts from its extensive collection, the museum constantly sets new thematic priorities and different ways to approach Picasso as an artistic figure.
The museum is located at Picassoplatz on the Königsstraße, once the most prestigious street in the old town. The exhibition rooms are spread across two historic buildings. The Druffel'sche Hof, built between 1784 and 1788, is one of the city's most important examples of Classical architecture. It was attached to the neighbouring Hensenbau building in the 1990s so it could be converted into a museum. The beautifully renovated façades of the two buildings that make up the Picasso Museum conceal an architecturally sophisticated building complex.
The museum space today houses more than 800 prints by Pablo Picasso, most of which come from the Huizinga collection. Thanks to the modern reconstruction of the buildings, Pablo Picasso Museum of Art is wheelchair accessible.
Opening hours:
Tuesday - Sunday and public holidays: 10:00 – 18:00


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My 1 Minute Goodnight Challenge that Made Me Happier
About a year ago I added one simple, quick exercise into my good-night routine and I can honestly say that I am happier and feel more complete because of it. It all started when I heard a quote at church. Henry B. Eyring said, "…I would ponder this question: 'Have I seen the hand of God reaching out to touch us or our children or our family today?' As I kept at it, something began to happen. As I would cast my mind over the day, I would see evidence of what God had done for one of us that I had not recognized in the busy moments of the day."
I went home and tried doing exactly that. Every night I try to find something specific to the day that I am grateful for that God has done for me. I took it a step further and also tried applying it to my husband. And these are the realizations I have made:
Finding Something Specific to Be Grateful for About My Family
Every night I try to think of something specific to thank my husband for before we go to sleep. Sometimes it gets hard and I have to be creative. But as I have tried to be creative, I have made an important discovery about praising and thanking others. Gratitude isn't simply about being thankful for things others do, but also for who they are as a person.
What I mean by it is this: it is easy to say thank you for doing the dishes or to express gratitude for help rocking the baby to sleep. But on days when my husband and I hardly see each other or we weren't our best selves I have started to express gratitude to him for things he simply is. For example, I'm thankful that he's silly and brings smiles to my face (sometimes accompanied by eye-rolling). I've become thankful that he works hard and is ambitious. I am grateful for his gentleness. I have always known these things, but it is different when I consciously remember it at the end of the day and express gratitude to him. I love him more and I feel happier. The unimportant things fall away, and I focus on what matters most.
Be Grateful for Who They Are rather than Only What They Do
I have come to the conclusion that being grateful for being rather than doing is probably far more important. It's nice to receive thanks when we offer a small act of service. But, it feels so much better when we receive thanks for being a supportive friend or a loving spouse or a hard worker. Praise that is about intrinsic traits rather than external things or acts is much more valuable in my opinion. It boosts our confidence and helps us know that we are making a difference to someone's life.
Praising Traits in our Children
Although my child isn't old enough to understand language in its entirety yet, I sometimes have started thanking her for her smiles or silly personality when we play baby games that we play together. The same warm effect comes over me and I find myself happier and less bothered by the unimportant things.
Of course I am far from perfect, but this praise is something I'm going to try to incorporate more and more into parenting. Praise is a core piece of numerous parenting programs. How great would it be that we not only thank our children for cleaning up their room and not back-talking, but also for who they are. Do we praise them enough for their creativity? Their kindness in including their younger siblings in play? Their sensitivity and their sweet spirit? Their ambition and fire in their spirit?
I know that sometimes it can be very difficult to focus on these things when somedays all we seem to hear are complaints and yelling. Some parenting programs tell parents to catch their children being good. I love that phrase. Everyone is good and does so much good. Sometimes we just have to change our focus from the negatives to the positives.
Being Grateful for a Higher Power
Of course, I also took the quote at face value and have made an effort to find something specific to be grateful for in my nightly prayers. I'm always grateful for things like family, but I have tried to be more specific in thanking God for something He blessed me with that day.
I love the example of Corrie Ten Boom. She was a woman living in Holland who headed an underground operation in Nazi occupied Holland to save Jews. She ended up going to a concentration camp and experiencing horrors that I can't begin to fathom. However, she held fast to her faith in Christ. She found the hand of the Lord in her daily life, even in the dark times. She found things to be happy about, even when there was really nothing to be thankful for. She understood this concept of looking deeper than the surface to just be grateful. She found the good in her surroundings and consequently lived a life of hope and joy afterward as she found forgiveness for the Nazis.
Reading her story in The Hiding Place had a profound effect on me. I could not do what she did, but I can start small. I can try harder to keep my bedtime routine of finding the hand of the Lord (or higher power that day) and finding goodness in the lives of my loved ones. Sometimes this requires creativity and looking past the easily seen contributions. But, I have found that as I have done so I have been happier and felt more complete.
*The Hiding Place is an Amazon affiliate link.
Eyring, Henry B. (2007) O Remember, Remember Retrieved from https://www.lds.org/general-conference/2007/10/o-remember-remember?lang=eng
Ten Boom, C., Sherrill, E., & Sherill, E. (1971). The Hiding Place. Grand Rapids, MI: Chosen Books.
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Motion Graphics
Lilypad Startup Promo
Storyboarding, illustration, and 2d animation. Made in collaboration with Adam Kennedy.
Azarga Promo
Color correction, logo animation and minor editing.
HPE Next Next: How lasers can reduce digital energy consumption
Photonic data transmission could cut data center energy use in half. That's the equivalent of decommissioning as many as 250 big power plants. Cut through obstacles with the power of light.
HPE Next Next: The quest to build a brain-like computer
Labs researchers are developing computers that work like the brain. They seek to realize the ancient dream of a thinking machine, and enable outcomes we can barely imagine. What can the human brain teach computer designers?
Style Guide Test 01
Pirate Santa Promo
Used Pirate Santa assets (Illustrated by Juan "Jun" Alvarado) to animate the Pirate Santa iPad app and promo. Created additional illustrations and animations for this sequence.
Animation and motion graphics reel (2012)
Pop-pop "Comic Promo"
Storyboarding, illustration, and 2d animation.