grafting frame holder
Vennalaganti P, et al. For plasma samples, raw mRNA capture sequencing reads were used to identify circRNAs based on back-splice junctions using CIRCexplorer2 (v2.3.3). (A) Plastic base mount cell cup. For circRNA detection, the CircExplorer2 manual was followed as described in the Methods section. In this study, we have identified several circRNAs in plasma of patients with EAC, HGD and NDB. Ferlay J, et al. Afterwards, variants were filtered differently depending on the tissue of origin. The concentration ranged from 16.3 to 2,210ng/l, with sample ID43_EAC (disease tissue) having the lowest concentration (Supplementary Table2). Description. To graft, lower the grafting tool behind the curve of the larva, maneuver the tool under the larva and the small pool of royal jelly, and gently lift and transfer the larva to the center of the cell cup (Figure 6). There are currently no studies reporting on mRNAs in plasma of patients with EAC, HGD or NDB. This helped identify and filter out variants introduced by misaligned reads. All code used for pre-processing mRNA and miRNA sequencing data is publicly available on GitHub (https://github.com/OncoRNALab/exRNAQC/blob/main/Preprocessing)61. 2021. Each cell bar holds 15 JZ/BZ cell cups for a total of 45 possible queen cells per grafting frame. Mechanisms that drive EAC development remain poorly understood. Annotation was based on GRCh38, UCSC Genome Browser (reference genome) and GENCODE v20, Ensembl 84 (reference transcriptome). Pleasantly surprised. The mean sequencing quality per base (raw data) of the small RNA sequencing data (tissue and plasma) is higher than 28 for the first 60bp in all samples (Fig. For instance, microRNAs (miRNAs) have been identified in tissue biopsies as potential biomarkers for EAC, HGD and NDB10. Regular price $13.95 Plastic Queen Cage. Esophageal cancer is the sixth most common cause of cancer-related death worldwide1. While several differential miRNA abundance patterns in EAC or NDB plasma have been reported21,5358, there is only one overlapping miRNA (miR-194-5p) among these studies54,56. Queen Bee Grafting Stand/Station and Frame Holder - Buzzbee These GSEA results (FDR<25%) are available in Supplementary Table3. Analysis of tissue and circulating microRNA expression during metaplastic transformation of the esophagus. Additionally, there is a low inter-observer agreement among pathologists in grading both low- and high-grade dysplasia, leading to over- and under-diagnosis7,8. Queen Catcher/Holder. Changes of microRNAs-192, 196a and 203 correlate with Barretts esophagus diagnosis and its progression compared to normal healthy individuals. The .gov means its official. Predictive value of microRNAs in the progression of Barrett esophagus to adenocarcinoma in a long-term follow-up study. When using the German grafting tool, surface tension from the priming liquid helps to transfer the larva from the grafting tool to the cell cup. JavaScript is disabled. 1 review. Double End Angled Stainless-Steel Grafting Tool for Queen Rearing. (careful edges are sharp) Ask us a question, when it's convenient for you. Information about cell builders can be found in the article Cell Builder Basics. BEEMAN. Grafting Frame - YouTube building the Grafting Frame (Multi use Frame) building the Grafting Frame (Multi use Frame) AboutPressCopyrightContact. A.P. Completely assembled 9 Deep frame designed to hold two JZBZ (JZ-BZ) Plastic Cell Bars or two Wood Cell Bars. Do not wait until the end of the graft or you will have a lower acceptance. (C) Egg. Blood samples were collected in a 6ml EDTA waste tube followed by a 9ml sodium citrate (3.2%) VACUETTE blood tube (Greiner Bio-One). document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); 2018 Barnyard Bees. Per patient, 1-7 variants were found, but no overlap was observed within a disease group or between groups. RNA concentration was measured with the Qubit 2.0 fluorometer (Thermo Fisher Scientific). In our analyses, no significantly differentially expressed circRNAs were identified (Table7). Too much liquid, however, can drown the larvae. Come join the discussion about breeding, honey production, health, behavior, hives, housing, adopting, care, classifieds, and more! (B) Cell bar with Nicot components. If using the Chinese grafting tool, simply deposit the larva and royal jelly in the center of the cell cup. Queen Rearing Grafting Frame - Barnyard Bees Spray with sugar solution. This comprehensive dataset includes 17 patients with EAC, HGD or NDB. With our analysis pipeline, no differentially abundant miRNAs between the plasma samples of the different groups were identified (Table7). A later subframe . 5.0 out of 5 stars Perfect. Holding up to a full-sized deep frame, which is positioned on the table surface is raised behind slightly to protect the brood on the reverse side. Zhang XO, et al. More to explore : Queen Bee Beekeeping Supplies, Bee Brush Beekeeping . Photos: Kate Anton, Penn State. Biomarker identification and trans-regulatory network analyses in esophageal adenocarcinoma and Barretts esophagus. Photos: Kate Anton, Penn State. Mallick R, Patnaik SK, Wani S, Bansal A. Liberzon A, et al. Stainless Steel Frame Bee Breeding Honeycomb Shelf For Bee Transferring https://www.amazon.com/innuoo-Micro+Intelligent+Microscope+HD+600x+Magnification, https://www.amazon.com/gp/product/B000784HKY/ref=oh_aui_detailpage_o03_s00?ie=UTF8&psc=1, http://m.ebay.com/itm/Rechargeable-%3Acd6660ee15a0a786c35161b6fff6e4ab%7Ciid%3A2. Queen Bee Rearing - Pt1 - Building the Grafting Frame A Systematic Review of Esophageal MicroRNA Markers for Diagnosis and Monitoring of Barretts Esophagus. Queen Rearing Frame Kit with JZBZ | Ango Apiculture For further downstream analyses, sample ID40_NDB was excluded due to the lower library yield (measured as described above) of the disease tissue sample (data not shown) and the lower percentage (68%) of reads with a quality score higher than 30, compared to all other tissue samples (85% on average). Deregulation of MYC is known to play a key role in the development of EAC36,37, indicating that tumor signal may be present in plasma. Profiling of circulating microRNAs in patients with Barretts esophagus and esophageal adenocarcinoma. Tomizawa Y, Wu TT, Wang KK. This holds the viewer on the longer top board so you can look at the whole frame. The queen lays eggs directly into a 10 x 11 grid of removable brown cell cups (Figure 8). Despite improved treatment strategies, the five-year survival rate remains unacceptably low (1025%)3,4. Would the closer focal length of the 5 diopter lens be a hindrance with your setup? Garzini | LinkedIn For this publication, raw data was pre-processed using in-house optimized pipelines (Biogazelle and Ghent University), resulting in 119 expression profiles: 34 mRNA and 34 miRNA expression profiles from healthy and disease tissue samples, 17 mRNA and 17 miRNA expression profiles from plasma, and 17 circRNA expression profiles (based on mRNA sequencing data) from plasma. Serum microRNA profiles as prognostic/predictive markers in the multimodality therapy of locally advanced adenocarcinomas of the gastroesophageal junction. Comparing EAC with NDB tissue samples, the WANG_BARRETTS_ESOPHAGUS_UP35 gene set was significantly negatively enriched in EAC (FDR<1%). For each patient, disease specific variants were identified using strict filtering as described in the methods section. The raw sequencing data available at EGA were not part of the peer-reviewed content of this manuscript. For example, comparing EAC tissue with healthy tissue the following relevant gene sets were significantly (FDR<1%) positively enriched in EAC: HALLMARK_EPITHELIAL_MESENCHYMAL_TRANSITION, HALLMARK_KRAS_SIGNALING_UP and WANG_ESOPHAGUS_CANCER_VS_NORMAL_UP35. Tissue and plasma samples were single-end sequenced with a 75bp read length on a NextSeq 500 (Illumina) instrument according to the manufacturers instructions. Get a sheet of foundation and cut a couple of pieces to fit either side. This was likely due to the low concentration (16.3ng/l) and low RNA quality score (3.4) (Supplementary Table2). De Preter K, Vandesompele J. This scale of queen production, along with the flexible behaviors inherent to honey bee colonies, are ultimately responsible for making honey bees the dominant pollinator in many landscapes. Before grafting, many beekeepers prime their cell cups with a small amount of royal jelly mixed with water, although water alone can be used. These authors contributed equally: Kathleen Schoofs, Annouck Philippron. Grafting Frame - BLB Honey & Beekeeping Supplies Excellent when working with the Technosetbee trifold frames in the Q8 Queen Mating Nucs. After using a grafting tool to remove larvae from brood cells, they are placed inside a queen cell cup which has been prefilled with royal jelly and is housed inside a cell bar frame. Mastering this technique takes practice and repetition. The beekeeper who only grafts occasionally will also appreciate this system. Subramanian A, et al. Queen Cell Production: Grafting and Graft-Free Methods Math EA, et al. Once this process is started, the queen cells should be handled gently, and care should be taken to avoid inverting queen cells again for the duration of development. These authors jointly supervised this work: Jo Vandesompele, Piet Pattyn, Katleen De Preter. The epidemic of oesophageal carcinoma: Where are we now? Time between blood collection and plasma preparation was less than 4h, except for sample ID2 (6h) and ID20 (7h). Mapped reads were annotated to mature miRNAs as well as other small RNAs, including tRNA, rRNA, sn(o)RNAs and piRNAs. The https:// ensures that you are connecting to the Acidic Bile Salts Induce Epithelial to Mesenchymal Transition via VEGF Signaling in Non-Neoplastic Barretts Cells. Move the frame not the viewer. contributed in the project planning, conceptualization, obtaining ethical approval for the study, sample collection, experimental work (plasma preparation, hemolysis measurement), and project management with Biogazelle. Current perspectives of mi-RNA in oesophageal adenocarcinoma: Roles in predicting carcinogenesis, progression and values in clinical management. Cielik M, Chinnaiyan AM. AboutPressCopyrightContact. In addition, results from variant- and fusion gene analysis are available as supplementary tables (Supplementary Tables4, 5). Matching tissue and blood samples were obtained from four patients with esophageal adenocarcinoma (EAC), five patients with high-grade dysplasia (HGD) and eight patients with non-dysplastic Barretts esophagus (NDB) (Table1). contributed in the data analysis (pre-processing mRNA- and small RNA sequencing data), data curation, and data interpretation. Mapping to the reference genome (Ensembl release 84) was performed with STAR (v2.5.1b) using the 2-pass mode and duplicates were removed with Picard tools (v2.7). We do not store credit card details nor have access to your credit card information. At least one of the tissue samples that was collected from the diseased tissue zone was sent for pathological investigation. On the diagonal line are the number of reported genes in each gene set. 10 Frame to 9 Frame Spacing Tool, Stainless, This frame perch is Beefy and very well made will stand up to heavy use A+. Experimental set-up and overview of the data. Regular price $5.50 German Queen Grafting Tool. The process of EMT enables cancer cells to enter the blood stream and form local and distant metastasis39. They are narrower than regular frames (facilitating movement in and out of the hive), and may be one of several designs. This article describes common grafting and graft-free techniques and equipment for producing queen cells in moveable cell cups (Figure 1). The basics of epithelial-mesenchymal transition. Amin M, Lam AK. Plasma was prepared by centrifugation at 1,800g for 10min (full break and acceleration). The cell bars fit into a specialized frame that can be introduced to a cell builder or starter, as detailed above. The pre-processed data is also uploaded in R2, allowing further exploration and visualization of the dataset. However, the overlap among these reported genes is limited. Grafting frame on a stand with a damp towel to maintain humidity. In total, 24 variants were identified in the plasma of two EAC patients, five HGD patients and four NDB patients (Supplementary Table4). 2021. A Data Transfer Agreement (DTA) and Data Access Agreement (DAA) will have to be signed in order for the data to be transferred (a template can be found in Supplementary File1). By subscribing toBuzzbee International text notifications, you agree to receive automated marketing text messages from us about our products and services at the phone number you provided when you subscribed, and that the messages may be sent via automatic telephone diallingsystem or other technology. Queen production allows beekeepers greater autonomy and independence, enabling individuals to better meet the goals of honey production, pollination, colony production, and genetic selection. If you are unable to perform a graft, you will find these replacement bar frames very handy. Differential MicroRNA signatures in the pathogenesis of barretts esophagus. VerticalScope Inc., 111 Peter Street, Suite 600, Toronto, Ontario, M5V 2H1, Canada. Standard Frame Holders that Suits wood & plastic frames available by Beequip. by using both miRNA and mRNA data, as demonstrated by this example. How about a frame holder that is good for an older back. When comparing EAC with NDB plasma, several relevant gene sets showed positive enrichment in EAC, including HALLMARK_MYC_TARGETS V1 and V2 (FDR<1%). In tissue samples, potential fusion genes were identified in all samples. The average number of reads for mRNA tissue and plasma samples throughout the pre-processing steps is shown in Table2. Delivered straight to your inbox (you can unsubscribe anytime). Suits wood and plastic frames. Queen Cell Cups for Base Mounting (50, 100 & 200 Packs) From $24.95. This technique allows beekeepers to create any number of queen cells that are easy to handle and transport. All components are reusable except brown cell cups. Clark RJ, Craig MP, Agrawal S, Kadakia M. microRNA involvement in the onset and progression of Barretts esophagus: A systematic review. Your payment information is processed securely. By gaining a deeper understanding in the different molecular subtypes, a more targeted treatment approach can be explored. EAC=esophageal adenocarcinoma, HGD=high-grade dysplasia, NDB=non-dysplastic Barretts esophagus, M=male, F=female, LGD=low-grade dysplasia, GEJ=gastro-esophageal junction. Craig MP, et al. ArrayExpress update - From bulk to single-cell expression data. Processed polyA+ RNA sequencing data of tissue samples from patients with esophageal adenocarcinoma or precursor lesions. Consent is not a condition of purchase. Lauri in your fourth picture down a close up of the cells there is a slightly curved egg or larvae at the 10 :00 position , is it a egg or larvae and do you have any good shots of the correct age that were looking for . This type of RNA has great potential as circulating biomarker because they are more resistant to RNA degradation by exonucleases due to their circular structure. Please enter your email address below to create account. A damp towel draped over the frame will keep the humidity high and should be used to cover the part of the frame that is not in use. (A) Brown cell cup and Nicot cup holder and socket. F.A.C. with a moist towel. An overview of all available data and access information is provided in Table3. The other disease tissue samples and healthy esopgahus tissue samples (collected from each patient) were preserved in RNAlater (Qiagen) at 4C and transferred to 80C the following day for long-term storage. The prefix p indicates histopathological staging of the resected tumor and y indicates that the patient received neoadjuvant therapy. (A) Grafting frame placement in cell builder. Technical validation of the data. Libraries were normalized and samples were pooled accordingly. Kit works great with our cell Grafting Frame. Maag JLV, et al. The pivot point is very close to the jaws which ensures a very good tight grip of the frame. Cell bars are put into graft-ing frames. for it, in 2012. Placing adequate shipping labels to Buzzbee to the address provided in step 1. I know you're organized enough to use a relatively new comb to graft from, and thus can cut away the cell walls so that you're able to easily get your grafting tool under the larva you're aiming to pick up. Tables5 and and66 show the overlap of differentially expressed genes (adjusted p-value<0.05) between EAC and healthy tissue from three large studies11,12,35 and our own dataset. Choose between Cell Bar Frame with 36 JZs BZs cell cups or Cell Bar Frame without cups. The relationship between miRNA and mRNA expression can thus be studied in our data. D.D.L. You may receive an additional text message confirming your decision to opt-out. In stock, 3 units. This is a holder to put your frame in while inspecting it. A capped queen cell and nurse bee. How to Set up Your Jenter Kit | Beespoke Info Excellent quality, sturdy and well made! Blum AE, et al. Report. $26.25. Clinical information of the 17 patients is available in Table1, including age at diagnosis, tumor stage and/or Barretts segment and follow-up information (if known). For each of the remaining variants, a BLAT (v3.5) analysis was performed to assess the quality of the reads contributing to the variant call31. Karczewski KJ, et al. P.P. Videos Help others learn more about this product by uploading a video! (a) quality plots of the RNA raw reads sequencing data: per base mean quality of mRNA tissue and plasma data (top row), and miRNA tissue and plasma data (bottom row); (b) hierarchical clustering of the mRNA plasma samples based on Pearsons correlation coefficient, generated in R2 (Euclidian distances, average linkage), where the R-value ranging from 1 to 1 represents the negative (1), positive (1) or no (0) relationship. Figure 4. Number of overlapping downregulated genes in EAC tissue compared to healthy tissue. Cell bars with plastic cell cups (60), used for rearing queens. Other useful tools include a grafting needle for precise transfers, and a royal jelly spoon for quick and easy harvest. . I did some grafting a couple years ago and everything was rushed with all the glass members a few of mine were accepted but now I'm not exactly sure what I'm looking for .I thought I would try the punch method this year . contributed in the project supervision, planning, conceptualization, data interpretation, and financial contribution. The Molecular Signatures Database (MSigDB) hallmark gene set collection. MicroRNA expression signatures during malignant progression from Barretts esophagus to esophageal adenocarcinoma. PDF THE GRAFTING METHOD OF CELL PRODUCTION - Wicwas Grafting Frame - YouTube Next, variants identified in the healthy tissue were subtracted from the tumor variant list to obtain a list of tumor-specific variants. Sole C, Arnaiz E, Manterola L, Otaegui D, Lawrie CH. 10 Frame Lyson Polystyrene Hive Components, 8/9 Frame Lyson Polystyrene Hive Components, Queen Bee Catching and Marking Tools and Cages, Honey Harvesting and Processing Equipment, Honey Heaters Dehydrators and Pollen Dryers, Bee Balm Pure and Natural Custom Honey Labels, Pure Honey River Scene Custom Honey Labels, Jar Top, Granulation & Other Honey Labels, Books, DVDs and Educational Tools for Beekeeping. Identification of fusion genes provides valuable insights in the development of EAC and can potentially be used as biomarkers for detection or therapeutic targeting. FOIA We rotate empty or food frames out of the central grafting box as necessary and use them to boost other NUCs or colonies. New frame holder for grafting | Bee keeping, Frame, Grafting - Pinterest Sequencing was done in two runs for all samples to obtain sufficient sequencing depth. Free delivery for many products! The store will not work correctly in the case when cookies are disabled. We offer quality beekeeping tools & equipment New Zealand wide. In addition, variants found in more than one gnomAD32 (v3.1) sample or having allele frequencies below 20 or above 80 percent were removed in the tissue data. Queen rearing schedule | Beesource Beekeeping Forums The upregulation of hsa-miR-194 and SHH, and downregulation of SUFU compared to healthy tissue is also observed in our NDB tissue data as well as in the EAC and HGD tissue samples (Figs. No overlapping fusion gene between disease tissue and plasma samples was observed. The clever design gives you quick and easy access to your cards . 2021. The Development and Validation of an Endoscopic Grading System for Barretts Esophagus: The Prague C & M Criteria. Beekeepers who wish to produce more than a handful of queens in a season typically do so by grafting. Frame Perch - Bee Frame Holder | Mann Lake Bee & Ag Supply Samples were paired-end sequenced with a read length of 275 base pairs (bp) on a NextSeq 500 (Illumina) instrument according to the manufacturers instructions. Ordering of mutations in preinvasive disease stages of esophageal carcinogenesis. Like mRNA, circRNAs have also not yet been reported in plasma from patients with EAC, HGD or NDB. There are many techniques to rear queens without grafting which work well for small scale production. Pre-processing of mRNA sequencing data of plasma and tissue samples included 3-end trimming, adapter removal and filtering (discard reads smaller than 20 nt) using Cutadapt (v1.18). Instead of taking over someone else's thread, I would like to share my grafting setup with everyone. " Find queen and place her on bottom deep and place queen excluder on top of bottom deep. Three variants are known tumor mutations according to the COSMIC database in prostate cancer (COSM5564582), cervix or biliary tract cancer (COSM5493837), or large intestine cancer (COSM5756079). In total, this comprehensive dataset consists of 102 sequenced libraries from 51 samples. Tympanoplasty is an inpatient procedure that is commonly performed under general anesthesia and sometimes local anesthesia. Bus P, et al. Dont forget to pick which cell bars you prefer to use with your Grafting frameWood Cell Bar or JZ-BZ Cell Bar Plastic and how many. Here's a super simple queen grafting frame build using only the parts provided with a standard frame and queen grafting cupholders. Frame grip. Similarly, two of the most reported downregulated miRNAs in EAC, HGD, and/or NDB are hsa-miR-203a-3p4651 and hsa-miR-205-5p4547,5052. If you do not do this, we will not be able to trace the goods to your account and may not be able to provide a refund or will reduce your refunds by the additional costs required to identify and trace the goods. Moreover, structural RNA differences can be identified in this dataset, including protein coding mutations, fusion genes, and circular RNAs. RNA extraction, library preparation and sequencing of all samples was performed by Biogazelle (Zwijnaarde, Belgium) as discussed in the next sections. All pre-processed mRNA, miRNA and circRNA expression data for tissue and plasma samples was also uploaded to the R2 Genomics Analysis and Visualization Platform (http://r2.amc.nl), an online genomics data visualization tool. Accepting all major credit-cards including Apple Pay, Google Pay and AfterPay. This method requires grafting. Choose between Cell Bar Frame with 36 JZ's BZ's cell cups or Cell Bar Frame without cups. Due to the ever-rising cost of outbound shipping, any order with backordered item(s) may be held until the backordered item(s) arrives. Circulating Serum Exosomal miRNAs As Potential Biomarkers for Esophageal Adenocarcinoma. Photo: Kate Anton, Penn State. Grafting is delicate work that requires patience, a steady hand and excellent vision. Clumpify (BBMap v38.26) was used for read duplicate removal for plasma samples only, due to the low RNA input. You must arrange and pay for the items to be returned to us within 30 days of receipt. We use the bamboo grafting tool to graftlarvae into queen cups on the cell bar frame. Workers freely enter and exit the box to care for the queen and young larvae. After using a grafting tool to remove larvae from brood cells, they are placed inside a queen cell cup which has been prefilled with royal jelly and is housed inside a cell bar frame. For the purpose of this study, two collections of the Molecular Signatures Database (MSigDB) were used: the hallmark26 and the C2 chemical and genetic perturbations gene sets. contributed in the project supervision, conceptualization, and financial contribution. Frankell AM, et al. Enjoyed seeing and reading your account, thanks! Nicot system viewed from the top and bottom. Results of expression and abundance analyses of tissue samples (19,734 genes and 676 miRNAs included) and plasma samples (11,255 genes, 457 miRNAs and 2,275 circRNAs included). If you have ever knocked over a frame propped against the hive, you know why you need this. Despite high resolution endoscopy and virtual imaging techniques, detecting dysplasia in a long segment of Barretts esophagus remains challenging. Message frequency is recurring. Transcriptional profiling suggests that Barretts metaplasia is an early intermediate stage in esophageal adenocarcinogenesis. Grafting a later front clip onto an early chassis replaces outdated suspension components to give your classic chassis better geometry for better road control and driving comfort. With a value proposition focused on anticipating and responding to the latest trends of the industry . The quality and the size distribution of the libraries was validated on the Fragment Analyzer (Advanced Analytical Technologies) and quantification was done using the Qubit fluorometer (Life Technologies). Further validation of these potentially relevant fusion genes is required. (sold separately) Complete Kit Includes Components to Produce 10 Queens: 1 Cell Grid 110 Brown Cell Cups 10 Hair Roller Cages Suits all the usual Langstroth size frames sold in NZ. It doesn't have remove-ab. 1Translational Oncogenomics and Bioinformatics Lab, Cancer Research Institute Ghent (CRIG), Ghent, Belgium, 2OncoRNALab, Cancer Research Institute Ghent (CRIG), Ghent, Belgium, 3 Department of Biomolecular Medicine, Ghent University, Ghent, Belgium, 4Department of Gastro-Intestinal Surgery, University Hospital Ghent, Belgium, 5Department of Oncogenomics, Academic Medical Center (AMC), Amsterdam, The Netherlands, 6Department of Gastroenterology and Hepatology, University Hospital Ghent, Ghent, Belgium, 7Department of Human Structure and Repair, Ghent University, Ghent, Belgium. Samples were paired-end sequenced with a read length of 275bp on a NextSeq 500 (Illumina) instrument according to the manufacturers instructions. Tool choice is subject to individual preference, and beekeepers often create their own from materials such as wire, a paper clip, or other common household items. Performance of RNA purification kits and blood collection tubes in the Extracellular RNA Quality Control (exRNAQC) study. Photo: Kate Anton, Penn State. We cannot accept returns on hazardous materials. (B) Grafting frame, with cell cups facing up, transported to the cell builder. Youll find some useful information to help you shop, different ways to get in touch, and our handy FAQs to help you find answers quickly. From all 51 samples, RNA was isolated that was used for mRNA (polyA+ and capture-based) and small RNA sequencing. We will provide you with the closest return shipping address, Upon receipt of the returned item, we will examine it and notify you via email whether you are entitled to a replacement, If so, we will send you a replacement and refund your cost of shipping.