The new PMC design is here! Learn more about navigating our updated article rfee. The PMC legacy view will also be available for a limited time. Federal government websites often end in. The site is secure. There are considerable differences in tumour biology between adult and paediatric cancers.

Identification of such cells hazel 4d linkedin free promote targeted therapies for paediatric solid tumours. Nevertheless, the cancer stem cell model has yet been applied for WT. In contrast, sorting according to CD, a marker proposed to identify embryonic renal progenitors and paediatric CICs Gillespie, ; Pode-Shakked et al, ; Ronconi et al,failed to generate a similar cell phenotype. Nevertheless, in vivo assays, e. Furthermore, tumourigenic favourable histology WT cell lines are not available.

Therefore, taking into account these inherent limitations of WT and the fact that cells within WT, as with other paediatric solid tumours, are less accessible compared to adult carcinomasmodel systems that allow for in vivo studies of WT at the single cell level are warranted. The generation of xenografts carrying cell-repopulating human developmental malignancies hazdl aggressive phenotypes may serve as a more reliable platform compared to cell-line-based xenografts or in vitro propagation for the discovery of CICs and future hazel 4d linkedin free therapies.

These numbers are in line with previous reports Wen et al, Xenografts are critical for this study because of the difficulty to routinely obtain paediatric primary solid tumours, which linkrdin less frequent than adult cancers.

Слова. citrix workspace password 1 password 2 это established 1st generation Xns from tissue samples obtained from eight different WT sources, we then analysed whether these Xns could be initiated gree propagated following dissociation of Xn tissue and injection of the derived hazel 4d linkedin free cell suspension.

Interestingly, we found that 1st generation Xns established from four WT sources W, W, W and W could be readily initiated and propagated in mice through the injection of 0. Having observed that p-WT Xn harbour enhanced WT-initiating activity, we next determined their characteristics in comparison with parental tumours. Histological examination of p-WT Xn W revealed all Freee tri-phasic components blastema, stroma, epithelium but with enriched blastema compared to parental tumour, indicating the generation of a blastema-predominant WT.

Interestingly, patients with recurrent frew WT have been classified at very high risk with low survival rates Reinhard et al, We freee immunostaining of p-WT Xn and parental WT for the following: WT1, a specific clinical diagnostic marker for WT; NCAM1, a putative marker for a tumour progenitor population previously shown in in vitro assays of primary cell cultures of WT Pode-Shakked et al, ; and Ki, a cell proliferation marker.

We further interrogated p-WT Xn at both gene читать полностью protein levels. In contrast, fold fewer CD negative cells were required to propagate and sustain late generation Xns of p-WT.

Since we observed that culture of WT cells derived from primary tumours abrogates their tumour-initiating capacity, we determined the effects of short-term hazel 4d linkedin free of p-WT Xn cells as monolayer or spheres on in vivo WT formation Supporting Information Rree S1, S2 and Tables S2 and S3.

Human CICs can hazel 4d linkedin free functionally identified by immunosorting according to hazel 4d linkedin free markers followed by transplantation in mice ljnkedin hazel 4d linkedin free dilutions Al-Hajj et al, ; Clarke et al, ; Lapidot et al, ; Pode-Shakked et al, We considered linkwdin expressed proteins only when at least a fivefold change appeared among samples.

These results implicated NCAM as a useful marker for prospective isolation of a cell sub-population enriched for WT-cancer initiating activity. Histology and immunohistochemistry revealed that these tumours recapitulated the tri-phasic WT phenotype and demonstrated widespread expression of the cell proliferation marker Ki Fig 3B. In the human foetal kidney, NCAM1, as 4 reported, was found to predominantly localized to the capping mesenchyme of the nephrogenic zone Roth et al, ab.

In contrast, ALDH1 localizes hazel 4d linkedin free mature tubules and is absent from the nephrogenic zone in the human foetal kidney Fig 3F upper panelwhile in WT it shares localization in both distinct cells of blastema as well as differentiated tubular elements Fig 3Flower panels. Nondenaturing PAGE and immunoblot analysis for the expression of proteasomal subunits 19s and 20s revealed overexpression of the subunits in comparison hazel 4d linkedin free both tumour xenografts generated from unsorted p-WT Xn cells and parental WT Fig 4A.

Importantly, the most significantly reduced microRNAs families: frree andas well as bazel, are related to control of epithelial cell differentiation and epithelial—mesenchymal transition EMT; Gregory et al, ; Li et al, ; Paterson et al, ; Peter, ; Wang et al, while miR has been linked to apoptosis, Ovcharenko et al, osteoblast likedin and was shown to be epigenetically silenced in oral cancers Kozaki et al, WT is currently treated both pre- and post-operatively with a combination of the two chemotherapic drugs vincristine and actinomycin D, to likedin other drugs may be added Green et al, Relapsing WT patients have reduced survival rates and often show resistance to conventional first line treatment Pinkerton et al, A combination of the topoisomerase II inhibitor, etoposide and cisplatin improves survival rates in these patients Sparano et al, WT cell death was significantly higher following application of lorvotuzumab-mertansine in comparison to treatment with the unconjugated antibody HuN or with saline only Fig 5A.

In accordance, there was a significant reduction in colony forming capacity of the WT cell cultures following treatment Fig 5Ca and fewer cells per colony Fig 5Ca lower panel and cast extension for windows 10. All groups were treated intravenously once a week ссылка на страницу three consecutive weeks and monitored for dree growth once-twice hazel 4d linkedin free week.

Tumour treated with lorvotuzumab-mertansine were completely eradicated following 3 hazel 4d linkedin free of treatment limkedin tumours treated with HuN or saline showed rapid tumour growth Fig 5Ea.

Comparison of mice weights revealed that while weight increased dramatically in both saline and unconjugated Ab treatment groups as a result of rapid tumour growth, immunoconjugate treated mice underwent weight loss during the first 2 weeks following total eradication of 4f hazel 4d linkedin free. However, in the 3rd week weights stabilized, indicating the lack hazel 4d linkedin free linoedin effects caused by the treatment Fig 5Eb. Intriguingly, of all mice treated with lorvotuzumab-mertansine two tumours relapsed after response.

Fdee generated WT xenografts that are continually initiated and propagated in mice by serial cell transplantation and recapitulated the primary tumour’s tri-lineage morphology. While libkedin able to form all lineages in fred tumour blastema, stroma and epitheliumclose examination of the propagatable xenografts revealed a shift towards highly vree blastemal predominant tumours that overexpressed renal stemness genes and those known to predict poor outcome in WT patients.

Thus, the establishment of human WT in immunodeficient mice selected and expanded a more aggressively malignant phenotype, reminiscent of the process of aggressive disease and relapse in patients. Indeed, divergent and high frequencies of tumourigenic cells with linkedun cancer-initiating activity have been suggested for certain malignancies Quintana et al, High ALDH1 activity levels have been used to better identify CSCs hzel a variety of tumours including linkedn Ma et al,head and neck Chen et al,colorectal Dylla et al, and breast Ginestier et al, tumour-initiating populations.

The localization of ALDH1 to a fraction of WT blastema cells but not to the undifferentiated blastema of the human foetal kidney located in differentiated structuresindicates that ALDH1 is likely to be associated with stem читать статью activity and CSCs, similarly to other cancers Ginestier et al, ; Ma hazel 4d linkedin free al, ; Pearce et al, ; Silva et al,rather than being a specific renal lineage blastema marker Metsuyanim et al, ; Roth et al, ab. These microRNAs have been closely hazel 4d linkedin free with 4r differentiation or maintenance and regulation of EMT in several malignancies Gregory et al, hazel 4d linkedin free Kong et hazel 4d linkedin free, ; Li et al, EMT has been shown to transform early tumours into /544.txt, metastatic and recurrent neoplasms and to be closely associated with a cancer stem hazel 4d linkedin free phenotype Mani et al, Our findings with respect to the CIC population may provide insight into libkedin cell lineage of origin for WT.

Given our observation that WT Xn CICs can give rise to blastemal, stromal and epithelial elements including tubular and glomeruloid bodies, indicative of abortive nephrogenesisone can assume a common undifferentiated renal mesenchymal cell of origin for all three cell hhazel.

Indeed, it is generally accepted that both the stroma and the nephron progenitors arise from the metanephric mesenchyme Kobayashi et al, ; Oliver et al, Alternatively, Hzzel CICs may relate to the more committed они autodesk autocad 2014 activation code generator free download Вам progenitor cells, a population shown to self-renew and differentiate exclusively to nephron epithelia throughout nephrogenesis, fulfilling the criteria of a true committed stem cell Kobayashi et al, Indeed, only later was the eradication of the cancer cell population they identify, proposed as a means to treat human frree Jin et al, ; Naor et al, This data should be interpreted in light of the fact that the IC50 of these drugs limits escalation of drug dosage in vitrowhile in vivo treatment might allow a higher dosage bearing an effect on the CIC population even by first-line chemotherapies.

Therefore, from a clinical standpoint, a combined regimen involving the eradication of the Hazel 4d linkedin free activity via targeting the NCAM molecule might prove useful in reducing chemotherapy toxicity in these patients, as well as in those that do not respond to conventional treatment or have a relapsing disease.

Our proof-of-principle experiments targeting this human cell fraction with the anti NCAM antibody-drug conjugate lorvotuzumab-mertansine, might ultimately pave the way for such an approach.

We propose that our findings may be relevant to other paediatric solid tumours representing a distinct biological entity, with linkexin similarities among themselves and marked differences linkwdin adult counterparts. The local ethical committee approved all studies linkediin informed consent was given by the legal guardians of the patients involved according to the hazel 4d linkedin free of Helsinki.

Briefly, primary WT tissue was hazel 4d linkedin free into 2—5 mm pieces and implanted subcutaneously in the back of the mice. In all the surgical procedures, mice were anaesthetized with i. Tumours were harvested approximately 3—6 months post implantation or when they reached a size of 1. WT xenograft derived cells thus generated were used for all subsequent assays. Engrafted mice were inspected bi-weekly for tumour appearance by visual observation and palpation.

Mice were sacrificed by CO 2 inhalation at a tumour diameter of 1. This was hazel 4d linkedin free at least three hazel 4d linkedin free. All animal experiments were conducted in accordance with the National Institutes of Health linkrdin for the linedin and use of animals and with an approved animal protocol from the Sheba Medical Center Animal Care and Use Committee. Linkedim cell suspensions from primary WT or WT Xn tissues were grown in two culture conditions as mentioned above: in SFM under vree attachment conditions where they formed free-floating spheres, or in SCM as adherent cultures for in vivo and in vitro assays.

WT, the most frequent tumour of the genitourinary tract in children, has served for decades as a general biologic model hazel 4d linkedin free study the intersection of cancer and development. Given the considerable differences in tumour biology between adult and paediatric cancers, the existence of cancer initiating and propagating cells in linkedi solid tumours including in WT is questionable. Identification of CICs in human Hazel 4d linkedin free is especially challenging due to minimal in vivo tumourigenicity of hazrl WT cells and lack of cell lines, hampering the ability to functionally define CICs via their enriched capacity to regenerate cancers in xenograft mouse models.

Finally, targeting of a cancer cell population enriched for CIC activity leads to dramatic tumour eradication or attenuation in human WT xenograft models.

This both provides molecular insights into tumour biology and opens practical new hzel to explore in the treatment of WT patients; Hazel 4d linkedin free though there have been successes in good prognosis WT treatment, our ultimate goal is to reduce the burden of conventional chemotherapy so as to decrease its long-term complications in this patient group. For the group of patients with relapsing bad prognosis disease treatments developed to eradicate CICs may have direct impact on morbidity and mortality.

Each mouse received bolus injections into the tail vein once a week for three consecutive weeks. Tumour dimensions were measured twice a week from the hazel 4d linkedin free conjugate injection until either hazel 4d linkedin free size reached mm 3 or mouse death. To determine the effect of lorvotuzumab-mertansine on HNWT and LNWT, mice bearing tumours were randomized into two groups, either treated with lorvotuzumab-mertansine or with saline as control.

During the experiment, mice hwzel were measured and documented. Statistical differences between additional data groups were determined using Student’s t -test. S 4 R. BD and NPS designed the experiments. ERG and YM provided the surgical samples. Such documents are peer-reviewed, but not hazfl or typeset.

They hhazel made available as submitted by the authors. Published online Dec Author information Article notes Copyright and License information Disclaimer. Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2. See » The stem and roots of Hazeo tumours » in volume 5 on page 4. This article has been cited by other articles in PMC. Abstract There are considerable differences in tumour biology between adult and paediatric cancers.

Keywords: cancer initiating cells, cancer stem cells, kidney stem cells, renal progenitor cells, targeted therapy. Human WT initiation and propagation in vivo rfee highly proliferating tumour cells Having established 1st generation Xns linkddin tissue samples obtained from eight different WT sources, we then analysed whether these Xns could be initiated and propagated following dissociation of Xn tissue and injection of the derived single cell suspension.

Open in a separate window. Figure 1. Staining demonstrates recapitulation of the original tumour by the WT Xn with an increase in the number of cells positive for NCAM and Ki67, markers of undifferentiated blastema and cell proliferation, respectively.

WT1, the diagnostic pathologic marker of WT, is expressed in both primary and Xn tumours on the same structures mainly blastema. In vitro growth conditions modulate WT-initiating activity Since we observed that culture of WT cells derived from primary tumours abrogates their tumour-initiating capacity, we determined the effects of short-term culture of p-WT Xn cells as monolayer or spheres on in vivo WT formation Supporting Information Figs S1, S2 and Tables S2 and S3.

Figure 2.

 
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Register | ProductionCrate.Packaging Machinery – Hazel 4D

 

And what have been the biggest opportunities? Ensuring that business can continue without interruption whatever the outcome of Brexit. What are the main trends you are expecting to see in the market in ? What technology is going to have the biggest impact on the market this year? You might also like. Does your supply chain need an upgrade? Leave a Reply Cancel Reply Your email address will not be published.

This website uses cookies to improve your experience. We’ll assume you’re ok with this, but you can opt-out if you wish. Hazel 4D specialises in helping companies improve pallet load stability in order to comply with the EU legislation around cargo transport safety.

Their clients include blue chip companies, specialist manufacturers and high-street retailers. They help customers optimise their pallet wrapping machine settings in order to apply their stretch film correctly and securely.

Antalis strengthens team with graduate recruits Nov 24, Key trends shaping the contract packing industry in Nov 24, Comments are closed. Carton erectors make forming and filling cardboard boxes quick and easy.

Designed to be combined with case sealers they can be integrated into existing production lines and are compatible with adhesive plastic machine tape or self- adhesive paper tape. Choose from semi-automatic or automatic machine options. Make a complete packing station by adding optional infeed and outfeed roller tables. All Comarme machines are compatible with our Outfeed Flexible Conveyor.

With our five-step consultation process, we discover your business needs, work with you to find the perfect packaging solution, implement it, and monitor its success. Reliable and quick, these new generation automatic and semi-automatic strapping machines will speed up your strapping operations.

Shop Machinery Shop friction weld tools. Ideal for intensive bundling and strapping operations where productivity is critical, this fully automatic strapping machine operates at high speeds.

 
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Contact Details – Fast and Easy Computers – Ample Seating’s – A Leading Office Furniture Store Online

 

Log in. Executive High Back Chairs 65 65 products. Ergonomic Office Chairs 72 72 products. Computer Tables 2 2 products. Conference Chairs 83 83 products. Add to wishlist.

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He knows how to perfectly deal with customer. If you guys are looking to buy office chair, I would suggest you to go with marvel chair as it’s very comfo and would be in your budget. Thank you so much shoeb. Genuine customer, Raj. Good services and good product I have ordered an ergonomic chair for work from home literally the chair is so good and quality is also so good I am very happy with ample seating delivery system chair has been delivered to my address safely and on time Highly recommended.

I had purchased Indian optima chair from Ample seatings and has been using it since close to 9 months now. The product is a little pricey, but it is worth every penny, this is the only thing that is making my long hours of office work manageable. I was referred to this by a friend who also made a purchase.

Overall the vendor is trustworthy and you can go for them. Menu Categories. Home Store Blog Contact Us. Shopping cart close. Nevertheless, in vivo assays, e. Furthermore, tumourigenic favourable histology WT cell lines are not available. Therefore, taking into account these inherent limitations of WT and the fact that cells within WT, as with other paediatric solid tumours, are less accessible compared to adult carcinomas , model systems that allow for in vivo studies of WT at the single cell level are warranted.

The generation of xenografts carrying cell-repopulating human developmental malignancies with aggressive phenotypes may serve as a more reliable platform compared to cell-line-based xenografts or in vitro propagation for the discovery of CICs and future targeted therapies. These numbers are in line with previous reports Wen et al, Xenografts are critical for this study because of the difficulty to routinely obtain paediatric primary solid tumours, which are less frequent than adult cancers.

Having established 1st generation Xns from tissue samples obtained from eight different WT sources, we then analysed whether these Xns could be initiated and propagated following dissociation of Xn tissue and injection of the derived single cell suspension.

Interestingly, we found that 1st generation Xns established from four WT sources W, W, W and W could be readily initiated and propagated in mice through the injection of 0. Having observed that p-WT Xn harbour enhanced WT-initiating activity, we next determined their characteristics in comparison with parental tumours. Histological examination of p-WT Xn W revealed all WT tri-phasic components blastema, stroma, epithelium but with enriched blastema compared to parental tumour, indicating the generation of a blastema-predominant WT.

Interestingly, patients with recurrent blastema-predominant WT have been classified at very high risk with low survival rates Reinhard et al, We performed immunostaining of p-WT Xn and parental WT for the following: WT1, a specific clinical diagnostic marker for WT; NCAM1, a putative marker for a tumour progenitor population previously shown in in vitro assays of primary cell cultures of WT Pode-Shakked et al, ; and Ki, a cell proliferation marker.

We further interrogated p-WT Xn at both gene and protein levels. In contrast, fold fewer CD negative cells were required to propagate and sustain late generation Xns of p-WT. Since we observed that culture of WT cells derived from primary tumours abrogates their tumour-initiating capacity, we determined the effects of short-term culture of p-WT Xn cells as monolayer or spheres on in vivo WT formation Supporting Information Figs S1, S2 and Tables S2 and S3.

Human CICs can be functionally identified by immunosorting according to surface markers followed by transplantation in mice in limiting dilutions Al-Hajj et al, ; Clarke et al, ; Lapidot et al, ; Pode-Shakked et al, We considered differentially expressed proteins only when at least a fivefold change appeared among samples.

These results implicated NCAM as a useful marker for prospective isolation of a cell sub-population enriched for WT-cancer initiating activity. Histology and immunohistochemistry revealed that these tumours recapitulated the tri-phasic WT phenotype and demonstrated widespread expression of the cell proliferation marker Ki Fig 3B.

In the human foetal kidney, NCAM1, as previously reported, was found to predominantly localized to the capping mesenchyme of the nephrogenic zone Roth et al, a , b. In contrast, ALDH1 localizes to mature tubules and is absent from the nephrogenic zone in the human foetal kidney Fig 3F upper panel , while in WT it shares localization in both distinct cells of blastema as well as differentiated tubular elements Fig 3F , lower panels.

Nondenaturing PAGE and immunoblot analysis for the expression of proteasomal subunits 19s and 20s revealed overexpression of the subunits in comparison to both tumour xenografts generated from unsorted p-WT Xn cells and parental WT Fig 4A. Importantly, the most significantly reduced microRNAs families: let7 and , as well as miR, are related to control of epithelial cell differentiation and epithelial—mesenchymal transition EMT; Gregory et al, ; Li et al, ; Paterson et al, ; Peter, ; Wang et al, while miR has been linked to apoptosis, Ovcharenko et al, osteoblast differentiation and was shown to be epigenetically silenced in oral cancers Kozaki et al, WT is currently treated both pre- and post-operatively with a combination of the two chemotherapic drugs vincristine and actinomycin D, to which other drugs may be added Green et al, Relapsing WT patients have reduced survival rates and often show resistance to conventional first line treatment Pinkerton et al, A combination of the topoisomerase II inhibitor, etoposide and cisplatin improves survival rates in these patients Sparano et al, WT cell death was significantly higher following application of lorvotuzumab-mertansine in comparison to treatment with the unconjugated antibody HuN or with saline only Fig 5A.

In accordance, there was a significant reduction in colony forming capacity of the WT cell cultures following treatment Fig 5Ca and fewer cells per colony Fig 5Ca lower panel and 5Cb. All groups were treated intravenously once a week for three consecutive weeks and monitored for tumour growth once-twice a week.

Tumour treated with lorvotuzumab-mertansine were completely eradicated following 3 weeks of treatment while tumours treated with HuN or saline showed rapid tumour growth Fig 5Ea. Comparison of mice weights revealed that while weight increased dramatically in both saline and unconjugated Ab treatment groups as a result of rapid tumour growth, immunoconjugate treated mice underwent weight loss during the first 2 weeks following total eradication of the tumours.

However, in the 3rd week weights stabilized, indicating the lack of toxic effects caused by the treatment Fig 5Eb.

Intriguingly, of all mice treated with lorvotuzumab-mertansine two tumours relapsed after response. We generated WT xenografts that are continually initiated and propagated in mice by serial cell transplantation and recapitulated the primary tumour’s tri-lineage morphology.

While being able to form all lineages in the tumour blastema, stroma and epithelium , close examination of the propagatable xenografts revealed a shift towards highly proliferating blastemal predominant tumours that overexpressed renal stemness genes and those known to predict poor outcome in WT patients. Thus, the establishment of human WT in immunodeficient mice selected and expanded a more aggressively malignant phenotype, reminiscent of the process of aggressive disease and relapse in patients.

Indeed, divergent and high frequencies of tumourigenic cells with enriched cancer-initiating activity have been suggested for certain malignancies Quintana et al, High ALDH1 activity levels have been used to better identify CSCs in a variety of tumours including liver Ma et al, , head and neck Chen et al, , colorectal Dylla et al, and breast Ginestier et al, tumour-initiating populations.

The localization of ALDH1 to a fraction of WT blastema cells but not to the undifferentiated blastema of the human foetal kidney located in differentiated structures , indicates that ALDH1 is likely to be associated with stem cell activity and CSCs, similarly to other cancers Ginestier et al, ; Ma et al, ; Pearce et al, ; Silva et al, , rather than being a specific renal lineage blastema marker Metsuyanim et al, ; Roth et al, a , b.

These microRNAs have been closely linked with epithelial differentiation or maintenance and regulation of EMT in several malignancies Gregory et al, ; Kong et al, ; Li et al, EMT has been shown to transform early tumours into invasive, metastatic and recurrent neoplasms and to be closely associated with a cancer stem cell phenotype Mani et al, Our findings with respect to the CIC population may provide insight into the cell lineage of origin for WT.

Given our observation that WT Xn CICs can give rise to blastemal, stromal and epithelial elements including tubular and glomeruloid bodies, indicative of abortive nephrogenesis , one can assume a common undifferentiated renal mesenchymal cell of origin for all three cell types.

Indeed, it is generally accepted that both the stroma and the nephron progenitors arise from the metanephric mesenchyme Kobayashi et al, ; Oliver et al, Alternatively, WT CICs may relate to the more committed nephron progenitor cells, a population shown to self-renew and differentiate exclusively to nephron epithelia throughout nephrogenesis, fulfilling the criteria of a true committed stem cell Kobayashi et al, Indeed, only later was the eradication of the cancer cell population they identify, proposed as a means to treat human malignancies Jin et al, ; Naor et al, This data should be interpreted in light of the fact that the IC50 of these drugs limits escalation of drug dosage in vitro , while in vivo treatment might allow a higher dosage bearing an effect on the CIC population even by first-line chemotherapies.

Therefore, from a clinical standpoint, a combined regimen involving the eradication of the WT-initiating activity via targeting the NCAM molecule might prove useful in reducing chemotherapy toxicity in these patients, as well as in those that do not respond to conventional treatment or have a relapsing disease. Our proof-of-principle experiments targeting this human cell fraction with the anti NCAM antibody-drug conjugate lorvotuzumab-mertansine, might ultimately pave the way for such an approach.

We propose that our findings may be relevant to other paediatric solid tumours representing a distinct biological entity, with great similarities among themselves and marked differences from adult counterparts. The local ethical committee approved all studies and informed consent was given by the legal guardians of the patients involved according to the declaration of Helsinki.

Briefly, primary WT tissue was cut into 2—5 mm pieces and implanted subcutaneously in the back of the mice. In all the surgical procedures, mice were anaesthetized with i. Tumours were harvested approximately 3—6 months post implantation or when they reached a size of 1. WT xenograft derived cells thus generated were used for all subsequent assays. Engrafted mice were inspected bi-weekly for tumour appearance by visual observation and palpation.

Mice were sacrificed by CO 2 inhalation at a tumour diameter of 1. This was repeated at least three times.

All animal experiments were conducted in accordance with the National Institutes of Health guidelines for the care and use of animals and with an approved animal protocol from the Sheba Medical Center Animal Care and Use Committee. Single cell suspensions from primary WT or WT Xn tissues were grown in two culture conditions as mentioned above: in SFM under low attachment conditions where they formed free-floating spheres, or in SCM as adherent cultures for in vivo and in vitro assays.

WT, the most frequent tumour of the genitourinary tract in children, has served for decades as a general biologic model to study the intersection of cancer and development. Given the considerable differences in tumour biology between adult and paediatric cancers, the existence of cancer initiating and propagating cells in paediatric solid tumours including in WT is questionable. Identification of CICs in human WT is especially challenging due to minimal in vivo tumourigenicity of primary WT cells and lack of cell lines, hampering the ability to functionally define CICs via their enriched capacity to regenerate cancers in xenograft mouse models.

Finally, targeting of a cancer cell population enriched for CIC activity leads to dramatic tumour eradication or attenuation in human WT xenograft models. This both provides molecular insights into tumour biology and opens practical new venues to explore in the treatment of WT patients; Even though there have been successes in good prognosis WT treatment, our ultimate goal is to reduce the burden of conventional chemotherapy so as to decrease its long-term complications in this patient group.

For the group of patients with relapsing bad prognosis disease treatments developed to eradicate CICs may have direct impact on morbidity and mortality. Each mouse received bolus injections into the tail vein once a week for three consecutive weeks.

Tumour dimensions were measured twice a week from the first conjugate injection until either tumours size reached mm 3 or mouse death. To determine the effect of lorvotuzumab-mertansine on HNWT and LNWT, mice bearing tumours were randomized into two groups, either treated with lorvotuzumab-mertansine or with saline as control. During the experiment, mice weights were measured and documented.

Statistical differences between additional data groups were determined using Student’s t -test. S and R. BD and NPS designed the experiments. ERG and YM provided the surgical samples. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors.

Published online Dec Author information Article notes Copyright and License information Disclaimer. Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2. See » The stem and roots of Wilms’ tumours » in volume 5 on page 4. This article has been cited by other articles in PMC.

Abstract There are considerable differences in tumour biology between adult and paediatric cancers. Keywords: cancer initiating cells, cancer stem cells, kidney stem cells, renal progenitor cells, targeted therapy. Human WT initiation and propagation in vivo via highly proliferating tumour cells Having established 1st generation Xns from tissue samples obtained from eight different WT sources, we then analysed whether these Xns could be initiated and propagated following dissociation of Xn tissue and injection of the derived single cell suspension.

Open in a separate window. Figure 1. Staining demonstrates recapitulation of the original tumour by the WT Xn with an increase in the number of cells positive for NCAM and Ki67, markers of undifferentiated blastema and cell proliferation, respectively.

WT1, the diagnostic pathologic marker of WT, is expressed in both primary and Xn tumours on the same structures mainly blastema. In vitro growth conditions modulate WT-initiating activity Since we observed that culture of WT cells derived from primary tumours abrogates their tumour-initiating capacity, we determined the effects of short-term culture of p-WT Xn cells as monolayer or spheres on in vivo WT formation Supporting Information Figs S1, S2 and Tables S2 and S3.

Figure 2. WT1, the marker used in clinical practice for histopathoplogic diagnosis of WT, is abundantly expressed in both WT xenografts on similar structures as in the primary tumour. Ki67 is more abundant in the WT Xn than in their primary source.

Figure 3. Experiments were repeated twice in triplicates. In primary WT, ALDH1 is expressed by some immature tubular structures and also by a small number of cells within the blastema. Figure 4. Proteasomal complexes were analysed by a nondenaturing page analysis and total proteasomal subunit protein levels and by b immunoblot analysis of polyacrylamide-SDS gel.

Suggested schematic representation of the regulatory pathways governing WT CIC phenotype and function. Figure 5. Cells derived from three different WT Xn W, W and W were exposed to the immunoconjugate, unconjugated huN antibody or saline.

Cell survival was significantly reduced in the lorvotuzumab-mertansine treated compared with HuN or saline treated controls. Cell survival was measured using an MTS assay. NCAM expressing cells were preferentially eliminated from the W cell population exposed to lorvotuzumab mertansin. Tumours treated with Lorvotuzumab-mertansine were completely eradicated following 3 weeks of treatment while tumours treated with either unconjugated Ab or saline showed rapid tumour growth p -values are 1. The authors declare that they have no conflict of interest.

Click here to view. Prospective identification of tumorigenic breast cancer cells. Phenotypic characterization of murine primitive hematopoietic progenitor cells isolated on basis of aldehyde dehydrogenase activity. Stem Cells.

 

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Kyle Stemmons’s email address and phone number

 
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Suggested schematic representation of the regulatory pathways governing WT CIC phenotype and function. Figure 5. Cells derived from three different WT Xn W, W and W were exposed to the immunoconjugate, unconjugated huN antibody or saline. Cell survival was significantly reduced in the lorvotuzumab-mertansine treated compared with HuN or saline treated controls.

Cell survival was measured using an MTS assay. NCAM expressing cells were preferentially eliminated from the W cell population exposed to lorvotuzumab mertansin. Tumours treated with Lorvotuzumab-mertansine were completely eradicated following 3 weeks of treatment while tumours treated with either unconjugated Ab or saline showed rapid tumour growth p -values are 1. The authors declare that they have no conflict of interest. Click here to view. Prospective identification of tumorigenic breast cancer cells.

Phenotypic characterization of murine primitive hematopoietic progenitor cells isolated on basis of aldehyde dehydrogenase activity.

Stem Cells. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med. Treatment of relapsed Wilms’ tumor with high-dose therapy and autologous hematopoietic stem-cell rescue: the experience at Children’s Memorial Hospital. J Clin Oncol. Identification, molecular characterization, clinical prognosis, and therapeutic targeting of human bladder tumor-initiating cells.

Aldehyde dehydrogenase 1 is a putative marker for cancer stem cells in head and neck squamous cancer. Biochem Biophys Res Commun. Cancer stem cells — perspectives on current status and future directions: AACR Workshop on cancer stem cells. Cancer Res. Prospective identification of tumorigenic prostate cancer stem cells. Multiple imprinted and stemness genes provide a link between normal and tumor progenitor cells of the developing human kidney. Colorectal cancer stem cells are enriched in xenogeneic tumors following chemotherapy.

Mapping the first stages of mesoderm commitment during differentiation of human embryonic stem cells.

Medical assessment of adverse health outcomes in long-term survivors of childhood cancer. Cancer stem cells and pediatric solid tumors. ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome.

Cell Stem Cell. Med Pediatr Oncol. Nat Cell Biol. The protein kinase Akt induces epithelial mesenchymal transition and promotes enhanced motility and invasiveness of squamous cell carcinoma lines.

J Clin Invest. Loss of miR inhibition of Suz12 leads to polycomb-mediated repression required for the formation and maintenance of cancer stem cells.

Mol Cell. Sci Signal. Odd-skipped related 1 is required for development of the metanephric kidney and regulates formation and differentiation of kidney precursor cells. Targeting the neural cell adhesion molecule in cancer. Cancer Lett. Targeting of CD44 eradicates human acute myeloid leukemic stem cells. Stem Cells Dev. Wilms tumour: prognostic factors, staging, therapy and late effects. Pediatr Radiol. Six2 defines and regulates a multipotent self-renewing nephron progenitor population throughout mammalian kidney development.

Epithelial to mesenchymal transition is mechanistically linked with stem cell signatures in prostate cancer cells. A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Beyond tumorigenesis: cancer stem cells in metastasis. Cell Res. Up-regulation of miR and let-7 by natural agents leads to the reversal of epithelial-to-mesenchymal transition in gemcitabine-resistant pancreatic cancer cells.

Aldehyde dehydrogenase discriminates the CD liver cancer stem cell populations. Mol Cancer Res. Frequent association of beta-catenin and WT1 mutations in Wilms tumors.

The epithelial-mesenchymal transition generates cells with properties of stem cells. Biochim Biophys Acta.

On this occasion, professional and Reunion Island Ideal for sporty, adventurous bon vivants. Wake up with the glow of the first rays of the sun over the mangrove forest. First a hearty breakfast with a view of the islands Nosy Carry out your projects in complete safety June 17, For all your credit or financing needs, we offer our services. Reliable and very secure with a good interest rate. The property is about 12 minutes drive from Bought 15th October at Conforma, guaranteed for 2 years.

Selling because we are moving. No delivery available. What is the net realizable value of this ac on December 31, ? Pitt pre delivery costs for Burr as an accommodation. On June 11, , what amount was received by Pitt form Bu remittance in full?

The payment due on the P, purchase was remitted on August The payment due on the P1,, purchase was remitted on August Under the net method and the gro method, these purchases should be included at what respective amount in the determination of cost of good. Net method Gross method a. Under the net method, the purchase discount is deducted from purchases regardless of whether taken or not Gross method Purchases 1,, Purchase discount taken 16, Net purchases 1,, Under the gross method, the purchases are recorded at gross and only the purchase discount taken is deduct from purchases in determining cost of goods available for sale.

Rabb Company records its purchases at gross amount but wishes to change to recording purchases net of pu discounts. Discount available on purchases for the current year totaled P, Of this amount, P10, i available in the accounts payable balance. The balances in the accounts as of and for the year ended Decem before conversion are:. Accounts payable at gross Discounts available in the accounts payable balance Accounts payable at net. Duke Company specializes in the sale of IBM compatibles and software packages.

It had the following tran one of its suppliers:. Purchases of IBM compatibles 1,, Purchases of commercial software packages 1,, Returns and allowances 50, Purchase discounts taken 17, All returns and allowances took place within purchase and prior to any payment on account.

Hero Company’s inventory on December 31, was P6,, based on a physical count of goods price cost and before any necessary year-end adjustments relating to the following:.

The invoice cost was P, The goods costing P, are properly included in the December 31, physical count because they ar shipped FOB shipping point only on January 7, picked up by common carrier.

The physical count conducted in the warehouse of Reverend Company on December 31, revealed merchandise with a total cost of P5,, However, further investigation revealed that the following ite were excluded from the count. It was not included in the inventory because it was marked «hold for shipping instructions». The customer was billed on that date and the machine was excluded from inventory although it was shipped on January 2, Physical count Inventory marked «hold for shipping instructions» Correct amount of inventory.

The inventory on hand on December 31, for Fair Company is valued at a cost of P, The follow items were not included in this inventory amount:. Item 1: Purchased goods in transit, shipped FOB destination, invoice price P30, which includes freight charge of P1, Goods are in transit. Baritone Company counted its ending inventory on December 31, None of the following items were included when the total amount of the ending inventory was computed:. The entity’s reported inventory before any corrections was P2,, What is the correct amount of inventory on December 31, ?

Sterling Comapany reported its year-end inventory at P7,, before the following adjustments:. The sale was properly recorded in A physical count on December 31, revealed that Joy Company had inventory with a cost of P4,,00 that the following items were excluded from this amount:. Mia Company submitted an inventory list on December 31, which showed a total of P5,, Taxes and duties have been paid on this shipment but the broker did not de until January 7, Delivered cost of the shipment totaled P, This shipment was not included i December 31, Shipment terms are FOB shipping point.

This items December 31, inventory by the entity. This was include in the y it was received only on January 5, The seller shipped the goods FOB destination. What is the correct inventory on December 31, ? Inventory per book Inventory transferred in delivery department Shipment consumed by bill of lading Goods in transit, purchased FOB destination Correct inventory.

The physical count conducted in the warehouse of Leila Company on December 31, revealed total cos However, the following items was excluded from the count:. The checks were mailed on January 10, Black recor payable at net amount. The invoice was recordedand paid January 3, B destination on December 20, from a vendor to Black were received January 2, was P, Accounts payable per book Undelivered entity checks Goods purchased and received on December 28, The undelivered checks should be adjusted as follows: Cash Accounts Payable 2,, 2,, Kew Company ‘s accounts payable balance on December 31, , was P2,, before considering the f.

The invoice cos On January 7, , Kew filed a P40, claim against the common carrier. A P70, credit memo was received a. Accounts payable per book Goods shipped FOB shipping point on December 22, and lost in transit Purchase returns Advance payment erroneously debited to accounts payable Adjusted accounts payable.

Kew Company shall suffer the loss of the goods in transit because the goods are shipped FOB shipping poi Company must file a claim against hte common carrier. Bakun Company began operations late in For the first quarter ended March 31, , Bakun made av. Total merchandise purchased through March 15, , recorded at net Merchandise inventory on December 31, , at selling price. No sales w What amount of cash is required to eliminate the current balance in accounts payable?

Aiza Company sells merchandise for P, to a customer on December 31, The terms of the sale is due in one year’s time. Lewis Company’s usual sales terms are net 60 days, F.

Sales, net of returns and allowan ended December 31, , before year-end adjustments. The goods were. The goods. Net sales per book Sales return Goods shipped on December 30, but recorded January 3, Goods shipped on January 3, erroneously recorded on December 30, Adjusted net sales.

Fenn Company had sales of P5,, during December On October 1,, Acme Company sold , gallons of heating oil to Kam Company at P30 per gallon on December 15, , and the remaining P50, gallons were delivered on January 15, What amount of revenue should Ac during ? On July 1,, Loveluck Company, a manufacturer of office furniture, supplied goods to Kaye Company f on condition that this amount is paid in full on July 1, Kaye had earlier rejected an alternative offer fr whereby it could have bought the same goods by paying cash of P1,, on July 1, What amoun should be respectively be recognized as sales revenue and interest income for the year ended.

Sales price 1,, Cash price – actual sales revenue 1,, Implied interest income , On July 1,, Kathleen Company handed over to a client a new computer system. The contract price for t the system and after-sales support for 12 months was P, What is the t revenue that should be recognized for ? Ilocos Company produced 80, kilos of tobacco during the season.

Ilocos sells all of its tobacco to customer which has agreed to purchase the entire production at the prevailing market price. Recent legislati assures that the market price will not fall below P per kilo during the next two years. The costs of sellin distributing the tobacco are immaterial and can be reasonably estimated.

Ilocos reports its inventory to expe exit value. During , Ilocos sold and delivered to the customer 60, kilos at the market price of P Ilocos sold the remaining 20, kilos during at the market price of P What amount of revenue sh Ilocos recognize in ? The remainder of the sales in of P1,, 20, x P50 is recognized as revenue in and not correction of revenue.

Beverly Company provides service contracts to customers for maintenance of their electrical system. On October 1, , it agrees to a four-year contract with a major customer for P1,, Costs over the peri of the contract are reliably estimated at P, What amount of revenue should be recognized for the ye ended December 31, ? The sale price of each piece equipment is P, Delivery of each piece of equipment is on February 10 of each year. In , the customer paid a P, down payment, and will pay P50, per year in and Collectibility reasonably assured.

Payment is to be m in two installments of P, each on December 1, and December 1, The goods are deliver on October 1, Collection is reasonably assured and the goods may no be returned. If the customer does not sell the goods to r customers by December 31, , the goods can be returned to Emco. The customer sells the goods to re customers on October 1, Marie Company, a distributor of machinery, bought a machine from the manufacturer in November for P10, However, Zoe had the right to return this machine to Marie if it was unable to resell the machine before expiration of the ninety-day payment period, in which case Zoe’s obligation to Marie would be canceled.

In Marie’s net sales for the year ended December 31, , what amount should be included for the sale of this machin. On January 1, , Bell Company contracted with the City of Manila to provide custom built desks for the The contract made Bell the city’s sole supplier and required Bell to supply no less than 4, desks and no 5, desks per year for two years.

In turn, the City of Manila agreed to pay a fixed price of P per desk. On December 31, , of these desks were segregated from the regular inventory and were accepted and awaiting pickup by the City of Manila. The Ci Manila paid Bell P2,, during What amount should Bell recognize as contract revenue in ? Delicate Company is a wholesale distributor of automotive replacement parts.

Inintial amounts taken from accouting records on December 31, are as follows:. Parts held on consignment from another entity to Delicate, the consignee, amounting to P,, were included in the physical count on December 31, , and in accounts payable on December 31, P20, of parts which were purchased and paid for in December , were sold in the last week of and appropriately recorded as sales of P28, The parts were included in the physical count on December 31, because the parts were on the loading dock waiting to be picked up by the customer.

Parts in transit on December 31, to customers, shipped FOB shipping point on December 28, , amounted to P34, The customers received the parts on January 6, Sales of P40, to the customers for the parts were recorded by Delicate on January 2, Retailers were holding P, at cost and P, at retail, of goods on consignment from Delicate, at their stores on December 31, Goods were in transit from a vendor to Delicate on December 31, The cost of goods was P25, The goods were shipped FOB shipping point on December 29, What is the correct amount of inventory?

What is the correct amount of accounts payable? What is the correct amount of sales? Held by consignees , , The terms of the invoice were orded in The goods cost P, The customer was expected ation on December 31, was. The inventory team ormed the supplier by long In addition, unt should Fenn report for net sales in. Forester Company on adoption of PAS 41 has reclassified certain assets as biological assets. The total value of the forest assets is P6,, which comprises:. Freestanding trees 5,, Land under trees , Roads in forests , 6,, In Forester Company’s statement of financial portion, what total amount of the forest assets shall be classified as biological assets?

The land under trees and roads in forests shall be included in property, plant and equipment. Colombia Company is a producer of coffee. The entity is cosidering the valuation of its harvested coffee beans. Industry practice is to value the coffee beans at market value and uses as reference a local publication «Accounting for Successful Farms». On December 31, , the entity has harvested coffee beans costing P3,, and with fair value less cost to sell of P3,, at the point harvest.

Because of long aging ang maturation process after harvest, the harvested coffee beans were still on hand on December 31, On such date, the fair value less cost to sell is P3,, and the net realizable value is P3,, What is the meaasurement of the coffee beans inventory on December 31, ? Fair value measurement stops at the point of harvest and PAS 2 on inventory applies after such date. Accordingly, the coffee beans inventory shall be measured at the lower of cost and net realizable value on December 31, The fair value less cost to sell P3,, at the point of harvest is the initial cost of coffee beans inventory for purposes of applying PAS 2.

The net realizable value of P3,, is the measurement on December 31, because this is lower than the deemed cost of P3,, Value of biological asset at acquisition cost on December 31, Fair valuation surplus on initial recognition at fair value on December 31, Change in fair value to December 31, due to growth and price fluctuation Decrease in fair value due to harvest.

Acquisiton cost – December 31, , Increase in fair value on initial recognition , Change in fair value in , Decrease in fair value due to harvest 90, Carrying amount – December 31, 1,, Change in fair value in , Decrease in fair value due to harvest 90, Net gain 10, Salve Company is engaged in raising dairy livestock.

Information regarding its activities relating to the dairy livestock is as follows:. Carrying amount on January 1, Increase due to purchases Gain arising from change in fair value less cost to sell attributable to price change Gain arising from change in fair value less cost to sell attributable to physical change Decrease due to sales Decrease due to harvest.

Carrying amount – January 1, Increase due to purchases Gain from change in fair value due to price change Gain from change in fair value due to physical change Decrease due to sales Decrease due to harvest Carrying amount – December 31, Honey Company has a herd of 10 2-year old animals on January 1, Oe animal aged 2.

No animals were sold or disposed of during the the fair value less cost to sell per unit is as follows:. What is the gain from change in fair value of biological assets that should be recognized in ? What is the gain from change in fair value due to price change? Price change 55 Physical Change Total gain from change in fair value Farmland Company produces milk on its farms.

Farmland Company own 5 farms and had a stock of 2, cows and 1, heifers. The farms produce , kilograms of milk a year and the average inventory held is 15, kilograms of mil However, on December 31, the entity is currently holding 50, kilograms of milk in powder. On Decem The biological assets are:.

Purchased before January 1, 3 years old Puchased on January 1, 2 years Purchased on July 1, old 1. The unit fair value less cost sell is as follows. January 1, 1-year old 3, 2- year old 4, December 31, 1-year old 3, 2-year old 4, 1. The entity has had problems during the year. Contaminated milk was sold to customers. As a result, milk consumption has gone down. The entiy’s business is spread over different parts of the country. The only region affected by the contamination was Batangas.

However, the cattle in this area were unaffected by the contamination and were healthy. What fair value of the biological assets on January 1, ?

Fair value – December 31, 14,, Fair value – January 1, 9,, Fair value – July 1, 2,, Increase in fair value 3,, Fair value of the milk produced , Gain from change in fair value 50, Total income , Inventories used , Staff costs , Depreciation expense 15, Other operating expenses , Income before income tax , Income tax Expense 55, Net income , On June 30, , all of Lin’s inventory was destroyed by fire.

The following figures pertain to Lin’s operations for the six months ended June 30, Net sales 8,, Beginning inventory 2,, Net purchases 5,, In the absence of any contrary statement, the gross profit rate is on sales.

On December 31,, a physical inventory revealed that the ending inventory was only P, Olivia suspects that some inv may have been pilfered by one of the entity’s employees.

On December 31,, what is the estimate cost of missing inventory? Sales 3,, Sales Returns 30, Net sales 3,, The sales discounts are ignored for purposes of estimating inventory under the gross profit method. On October 31,, a flood at Pamel Company’s only warehouse caused severe damage to its entire Based on the recent history, Pamela has a gross profit of 25 percent of sales. A physical inventory disclosed usable damaged goods which Pamela estimates can be sold for P70, Using the gross profit method, what is the estimated cost of goods sold for the ten months ended Oct a.

Like sales discounts, sales allowances are ignored in determining net sales under the gross profit m. The following data are available for nine months September 30, Inventory at January 1 1,, Net purchases 6,, Net sales 7,, A physical inventory disclosed usable damaged goods which can be sold for P , What is the estimated cost of goods sold for the nine months ended September 30, ? On the night of September 30, , a fire destroyed most of the merchandise inventorry of Sonia Co All goods were completely destroyed except for partial damaged goods that normally sell for P,0 that had an estimated net realizable value of P25, and undamaged goods that normally sell for P6 The following data are available:.

No inventory had been taken on December 31, The following profit and loss data are available:. Sales, net and 7,, Cost of sales: Inventory – January1, , Net purchases and 5,, Goods available for sale 6,, Inventory – December 31, 1,, 5,, Gross Profit 2,, Beyonce Company sells merchandise on a consignment basis to dealers.

The following consignment activities occured during Manufacturing cost of goods shipped on consignment Sales price of merchandise sold by dealers Payments remitted by dealers after deducting commission.

Steven Company began operations in For the year ended December 31, , Steven made ava the following information:. Total merchandise purchases for the year 7,, Merchandise inventory on December 31 1,, Collection from customers 4,, All sales are on a credit basis and all receiva collectible.

What is the balance of accounts receivable on December 31, ? On December 31, , Empress Company had a fire which completely destroyed the goods in proce inventory.

After the fire a physical inventory was taken. The raw materials were valued at P,, the finished goods at P1,, and supplies at P,0 on December 31, The inventories on January 1, consisted of the following:. What is the estimated cost of the goods in process on December 31, that wre completely destroyed by fire? In conducting an audit of Romy Company for the year ended June 30, , the entity’s CPA observe the physical inventory at an transaction date, May 31, The following information was obtained Inventory, July 1, , Physical inventory, May 31, , Sales for 11 months ended May 31, 8,, Sales for year ended June 30, 9,, Purchases for 11 months ended May 31, 6,, Purchases for year ended June 30, 8,, Shipments received in May and included in the physical inventory but recorded at June purchases b.

Shipments received in unsalable condition and excluded from physical inventory. Credit memos had not been received nor had chargebacks to vendors been recorded Total at May 31, Total at June 30, including the May unrecorded Chargebacks c.

Deposit made with vendor and charged to purchases in April, Product was shipped in July, Deposit made with vendor and charged to purchases in May, Product was shipped FOB destination, on May 29, and was included in May 31, physical inventory as goods in transit.

Through the carelessness of the receiving department a June shipment was damaged by rain. This shipment was later sold in June at its cost of.

What is the cost of goods for the month of June ? What is the June 30, inventory? Physical inventory Purchases up to Purchas e May 31, May 31, June 30 Balances , 6,, 8,, a – 75, – b – 10, 15,0 c – 20, 20,0 d 55, 55, – Adjusted , 6,, 7,,.

On April 30, , a fire damaged the office of Amaze Company. Accounts receivable , Inventory – January 1, 1,, Accounts payable , Sales 3,, Purchases 1,, Accounts payable as of March 31 , April merchandise shipments 80, Expenses , Deposits during the same period amounted to P, which consisted of collections from customers with the exception of P20, refund from a vendor for merchandise returned in April.

Customers owed another P60, that will never be recovered. Of the acknowledged indebtedness, P40, may prove uncollectible. The balance of the inventory was a total loss. On December 31, , the following information was available from Huff Company’s accounting records:.

Cost Retail Inventory, January 1 , 1,, Purchases 4,, 5,, Additional markups – , Available for sale 4,, 7,, Sales for the year totaled P5,, Markdowns amounted to P70, Under the approximate lower of average cost of market retail method, what is the inventory on December 31, ? The approximate lower of average cost or market retail method is the same as the consevative or conventional retail approach.

Data relating to the inventory computation on December 31, are as follows:. Caramel Company uses the average retail inventory method. On December 31, , the following information re.

Note that the sales discount and sales allowance are ignored in determining the net sales under the retail method. Diane uses the average cost retail inventory method in estimating the value of its inventory.

What is the estimated cost of inventory on December 31, ? The following informatio for the current year:.

Cost Beginning inventory and purchases 6,, Net markups Net marktowns Sales. On January 1, the stock inventory of Ron Company was P1,, at the retail and P, at cost. During the current year, the entity registered the following purchases:.

To meet price competition 50, To dispose of overstock 30, Miscellaneous reductions , During the current year, the selling price of a certain inventory increased from P to P This additonal markup applied to 5, items but was later canceled on the remaining 1, items. Pro For Individuals. Payments are securely processed and can be canceled at any time. Enterprise For Teams and Companies.

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By building trust, you foster psychological safety. It can be a simple document or a presentation. After all, our personalities and preferences are not fixed. Here are six prompts to get you going. Ask them if anything is unclear or seems to be missing.

Then, revise your first version based on their comments. Some people decide to share their personal user manual publicly, others only share it with their teams.

There is no obligation. Share it with as many people you feel comfortable sharing it with. But the more people are aware of the way you work, the easier it will be to work with them.

Remember that a personal user manual should be a living document. Of course, the document will lose some of its usefulness if you keep on updating the content every week—your colleagues may get confused as to how best to work with you.

But you could take some time to reflect on your personal user manual as part of your annual performance review. Have your values or working style changed? Any big shifts? If you need some inspiration, here are some interesting personal user manuals.

Feel free to get creative with the questions, as long as all the essential information is included in the final version. By fostering psychological safety, being open as a leader is a catalyst for trust and performance. Do you have a personal user manual? Maker Mind is a weekly newsletter with science-based insights on creativity, mindful productivity, better thinking and lifelong learning.

Ness Labs provides content, coaching, courses and community to help makers put their minds at work. Apply evidence-based strategies to your daily life, discover the latest in neuroscience research, and connect with fellow mindful makers. I guess it has to do with the medium. Not a person. I was probably replying right before a meeting or in between working on projects.

That was my fault for choosing to respond then. If there is a big issue, we will discuss it in person — email is typically not the best communication for big-ticket problems. Whatsapp for quick communication. Google Docs. Loom for quick video feedback.

Maybe management will take notice and use it and make it a new part of their company strategy. Disclosure: this article might contain links to the resources discussed.

Some of the links are affiliate links, meaning that I will make a small commission if you purchase a product or service by using the link. More info in my privacy policy. Brooks is a firm believer in creating multiple streams of income and creates content here on BrooksConkle.

Full inbox? I feel you. Follow me on YouTube if that’s easier. Table of Contents. Brooks Conkle. Recent Posts. Find the adventure in this category. Software This is software and tools that I recommend to help readers along their journey.

 
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Personal user manual.Fostering psychological safety with a personal user manual

 

– Не знаю; может, и эмоциональное впечатление может произвести лишь ее отсутствие. Джезерак выслушал ее рассказ без видимых эмоций. Как будто некая сила, Элвин, что кто-то позволял себе рассматривать жителей Диаспара как какие-то существа низшего порядка.

 
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How to foster psychological safety with a personal user manual

 

Jenna Bunnell is the Senior Manager for Content Marketing at Dialpad, an AI-incorporated cloud-hosted unified communications and call center IVR systems that provide valuable call details for business owners and sales representatives.

Find her on LinkedIn. Personal user manuals are indispensable tools when it comes to working with new employees and perfecting your new employee onboarding process. They are written guides that explain to others how best to work with you. They can either be targeted towards people working under you or people working with you.

Personal user manuals are full of information for other employees. They promote team collaboration by making others aware of your personal working style, and allowing them to work best with this. A personal user manual can be used instead, saving time, and allowing effective collaboration regardless of different time zones. You may well be wondering: How on Earth do I get started with such an undertaking? Well, here are some steps you can take to create and maintain a solid personal user manual.

This post for Process Street will outline the 10 focus areas to include in your personal user manual:. So, consider an online service that keeps track of updates, like Google Docs. What would be essential to know if someone came into the office now? Would they need to know, for example, whether you use VoIP versus landline? Outlook versus Gmail? The first thing you should say in the document is something to put any new employee at ease.

Welcome them, and explain what a joy it is to have them working with you. You should also include basic information about you, your career, and your life. Mention what you enjoy most at work, as well as what you enjoy most outside of work. Overall, as a boss, you may be relaxed, strict, problem-solving, vision-based, or anything else. Try your best, in the introduction, to give an overview of your management style and the benefits of this style, as well as your organizational priorities.

Go into your background and your various skills, casting light on how you work in an office setting and what your ethos is.

Although the employee will have already seen the job description, they will likely want to know more about the specifics. This includes information about your role and how the two roles will fit together.

What tasks do you expect them to do, and which should be left to you? How do they fit in with the other employees at the company? Are they more involved in app testing , ecommerce analysis, or customer service? Do these things ever overlap? Teamwork makes the dream work, as they say. Ensuring that cooperation is possible from the start will allow for a seamless transition once you start working together.

From the moment you enter the office, jot down what you do and when, and more importantly, how you do it. What time do you usually get into the office? Note that down. Make notes about the various employees in the office, the structure of the office, your computer setup, where the toilets are.

You might also like to note all of the computer programs you use — ecommerce analytics tools , administrative software, customer relationship programs, etc. Put a notepad and pen in your pocket and continue for the whole day. Then, write a walkthrough of everything that your reader can expect. You may also want to write down details about what the employee will be doing all day, perhaps even modeling this on some of your current workers and how you see this new role fitting in.

One of the main things that readers will want to know about is your work schedule. What do you do all day, anyway? More specifically, what do you do when you start your day? What do you do for lunch? This may also include information about regular meetings, business trips, and even planned holidays throughout the year. Make sure there are no surprises for your staff, by being upfront about how the months will progress, and any pressing issues that you have coming up.

By asking yourself the following questions, you can get a better idea of what you could add to your personal user manual:. Usually, your colleagues are the ones who see you and witness your work every day, and as such, they will be able to provide unbiased feedback about you and the way you work.

However, make sure to ask the right colleagues in your research. To further increase the likelihood of honest responses, you might consider sending around an anonymous online survey or creating a suggestion box. Ask colleagues what you think people should know about you as a boss and how to work in the office. Most managers know not to micromanage , not to pester, and not to have sky-high expectations, but when it comes down to not doing these things in practice, it can be a different story.

Considering these when writing your personal user manual will help you to write with empathy. The manual is also a great tool that will provide support to your future employees, as a resource that they can come back to for reference. Think about what issues may arise in light of some of these difficulties, including those highlighted by your employees.

Reflecting on past feedback from colleagues will help you to create a picture of your work personality. Inevitably, other personality types may conflict with this.

The manager-employee relationship is a complicated one, and one that is inherently hierarchical. In general, employees cannot do much to change the dynamic of the relationship, due to the power imbalance that will always be there. Before sitting down to write the document, get in the mindset of being able to manage the relationship in a healthy way. Just like when you work with the public and need to consider voice of customer approaches i.

You want to write your personal user manual in a friendly and humble way, rather than putting too much blame and responsibility on the recruit, a mistake often made by managers when writing manuals. After all, the worker is there to be managed, not to read a manual and be quizzed on it. Rather than providing instructions in your manual, get into the mindset of being welcoming and giving helpful tips and advice. The below image illustrates how the manager-employee relationship is made up of positive steps taken by the manager.

Conflict resolution is very important in the workplace. You can nip any conflict in the bud by recognizing what could happen and explaining it in detail in the personal user manual. Every workplace should have a conflict resolution process, including who to go to, how to report an issue, and what should be done to resolve it.

You can easily make conflict resolution a part of the manual and introduce employees to the process before anything comes up, improving employee relationships and team collaboration. Keeping a personal user manual is a constant process, as the manual will require constant updates based on how you, your routine, and the business change. As you get to know the employee better, and they become part of the furniture, you may also consider asking them to help out by adding to the personal user manual as they see fit.

With a constantly updated document, current employees will also be able to go there for news and changes to processes, as well as reminders of the way things work. You could even enlist one of your employees, who would like to, to act as a personal user manual update officer, constantly adding updates as they, their colleagues, and you think of them. Ask for feedback once your employees become familiar enough, making sure to collect data on whether the written guide helped them.

You can ask the other, established employees in the office whether or not the introduction of the personal user manual helped them, how much, and in which ways. Another good way to measure how well your manual is working is by assessing morale.

When coming up with alternatives, you could brainstorm within the team, but you may also wish to consult a professional, such as an account executive. However you choose to do it, implementing a personal user manual really could help supercharge team collaboration.

Leks Drakos, Ph. Contact Sales. Try it free. Human Resources. This post for Process Street will outline the 10 focus areas to include in your personal user manual: Choose a format for your personal user manual Personal user manual Introduce yourself Outline your expectations of your team as a manager How to work with me: The essentials Scheduling: When are you accessible?

Source Personal user manual Introduce yourself The first thing you should say in the document is something to put any new employee at ease. Outline your expectations of your team as a manager Although the employee will have already seen the job description, they will likely want to know more about the specifics. When are you accessible? By asking yourself the following questions, you can get a better idea of what you could add to your personal user manual: What motivates me?

How can employees succeed with me? What might employees not realize about me? Source Conflict resolution and you: Everything you need to know Conflict resolution is very important in the workplace.

Source Every workplace should have a conflict resolution process, including who to go to, how to report an issue, and what should be done to resolve it. Personal user manual: The next iteration Keeping a personal user manual is a constant process, as the manual will require constant updates based on how you, your routine, and the business change.

Let us know! Leks Drakos Leks Drakos, Ph. Leave a comment Your email address will not be published. Required fields are marked. Take control of your workflows today Get started Contact Sales. Use cases. For customers.

 
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What advantages do you get from our Achiever Papers’ services? The MII naming convention may be thought of as a generic brand name that is also used for products i. Each of these will be detailed further in the next section. Similarly, the system-level requirement of the transmission medium, such as copper cable and fiber-optics, needs to be considered.

To demonstrate this point, you should check the product page of the selected PHY in the design example, which is the KSZ There is no need to mention the MDI side as this is relatively clearer when it comes to selecting the correct PHY, with the choice between fiber and copper cable interfaces. Reading and understanding any gigabit PHY datasheet may not appear that easy at first glance since there will be a lot of standard-defined properties listed in the features section.

We will try to describe some of these briefly and if you feel some extra information is needed for your specific application, just google the appropriate keyword:. Gigabit Ethernet uses PAM-5 modulation that uses five voltage levels and encodes two bits per clock cycle using four different voltage levels in each pair; the fifth voltage level is used for error-correction.

So, using the same Baud rate and clock frequency as the Fast Ethernet, the Gigabit Ethernet uses all available resources more efficiently and increases the link speed, all the while keeping within the certified limits of the relatively cheap Cat5 cable rather than needing to use more expensive higher category cables.

Also, additional clock references such as 2. It is always a good idea to double-check the datasheet to see if it has a built-in crystal driver that makes it able to use crystal. Generally, the accuracy requirements are advised to be better than 50 ppm, and utilizing an oscillator may make the layout easier. Again, it is a trade-off for designers in terms of price, stability, and layout effort.

You have to be careful to check the crystal load capacitance if you select this option. It is strongly recommended to check the datasheet carefully for the strap options since these are vendor-dependent and may be subject to change between each device.

The crucial point here is to adjust the required reset time for the strap pins to settle at the desired voltage level, which is easily adjusted using an RC delay circuitry. Another point related to PHY selection is to check whether it has internal termination resistors or not.

The MDI uses balanced differential pairs, and so if the PHY does not have on-chip termination resistors, parallel split termination preferred to filter common-mode noise must be added to the board. Similarly, the xMII interface should have series termination resistors, either on-chip or on-board. As briefly mentioned, while describing the use of the magnetics center-tap, there are two types of line-driver available for gigabit Ethernet: Current Mode and Voltage Mode.

The designer should check the PHY line driver for a magnetics center-tap and split termination center-tap connection. Since voltage-mode drivers have various advantages over current-mode, nowadays, this type of line driver is more prevalent among devices.

However, the designer should still be aware of the current-mode line driver requirements for different design aspects. Other analog, digital, and IO supplies are usually selectable from 3.

To allow single-supply operation, the device may have an integrated LDO controller e. If the board already has a separate 1.

As FET selection is strictly related to the controller, designers should follow the recommendations within the datasheet for FET specifications. After selecting the correct PHY to fulfill the requirements and following the above recommendations, the schematic design is fairly standard regardless of the chosen device and follows these steps:.

Some explanatory notes and device-specific pin connections are given inside the schematic. Transmitted and received signals must be synchronized using clock signals. It is vital to bear in mind that evolving technology not only increases bandwidth requirements but can also result in a lot of interfaces in use at the same time.

This is why having at least one GPIO pin may be valuable to future-proof the overall design. Signal descriptions are given in Table 2 below. For gigabit communications, data is clocked on both the falling and rising edges of the MHz clock, which results in a halving of the data signal count.

Both of these are source-synchronized clock signals, and they use both the falling and rising edges of the clock, which makes timing more critical. If it is not supported by one or both devices, then the delay must be applied as part of the PCB layout by using correctly designed serpentines, as shown in Figure 15 below.

Always double-check the naming conventions before designing the layout. Single-ended parallel bus topologies need series termination on the driver side to match both the output driver impedance and the line characteristic impedance, to prevent reflections and EMI problems. Designers should check the PHY and MAC datasheet for the presence of internal termination resistors, and if they do not exist, they must be placed on-board. Generally, values between 20 ohms and 40 ohms will work, but some trial and error may be needed to get the best performance.

SGMII reduces the pin count and increases the speed, but the downside is that the layout is more complicated than for the xMII methods. Most SerDes high-speed interfaces require capacitive coupling to prevent receiver-transmitter common-mode voltage mismatches. It is recommended to have at least place holders for nF series capacitors close to the TX side of the SGMII pairs, along with parallel termination resistors according to the differential pair impedance usually ohms or ohms..

This interface is similar to the I2C bus and is used by upper-level devices such as the MAC to acquire the PHY status and to program the PHY registers to tune changeable run-time parameters like clock settings and fix-up routines. It is worth mentioning that you may not need to add multiple Ethernet PHY and MAC devices on your circuit board unless there are strict requirements for the physical separation of the interfaces.

The best practice is to use isolation transformers for all the PHY interfaces located on the board, similar to the RJ connector operation. However, this method is expensive and uses up a lot of board space. Instead, all the pairs are capacitively coupled using series nF capacitors. Although it is not guaranteed to work over long distances, in theory, it works very well over relatively short distances. After reading hundreds of datasheet pages, you have a perfectly designed schematic that meets all the requirements and suggestions the manufacturers have suggested – however, all that effort can easily be ruined or have degraded performance due to fundamental layout failures.

For the design of a Gigabit Ethernet interface, there are impedance controlled differential and single-ended signals to be considered, as well as some length matching and maximum length limitations. Most of the time, these requirements are automatically fulfilled by the sensible placement of the components unless the designer tries to override this approach. The problem is that if general layout rules are not obeyed such as not using solid reference planes for Ethernet impedance controlled traces , it is a waste of effort to strictly match trace lengths or keep them below the maximum length limits.

Therefore, we will briefly describe the generic high-speed layout rules before specific gigabit Ethernet layout requirements are discussed to provide a foundation for the more specific requirements. High-speed switching digital ICs demand transient currents. The main rule is to place bypass capacitors as close as possible to all the supply pins, with at least one 10 nF and nF capacitor for each pin.

For multi-layer boards , there are separate power and ground planes, and so vias will inevitably be used in the path used to supply power. Since vias also have an inductive component, no via should be used between a bypass capacitor and its associated supply pin. This rule is illustrated in Figure 20 below.

The basic rule for all electronics is that the current flowing in the circuit always returns to its source. So, there should always be a return path for the signals, and this return path will form a loop antenna with the outbound signal path. Based on both the theory and experimental evidence for high-speed signals, the current return path will follow the trace on the layer that is beneath it.

In other words, its reference plane. Keeping a solid reference plane below any high-speed signal routing will minimize the loop area and prevent any impedance discontinuity. If for any reason, plane voids are created beneath a high-speed trace, stitching capacitors should be used to create a return path. The use of stitching capacitors is also recommended if the power plane is also the reference plane for a high-speed signal that creates a return path to the current source.

These rules are illustrated in Figure 21 below, showing bad practices on the left and good practices on the right. This does not mean that it is impossible to use a two-layer PCB for a gigabit Ethernet interface. Each trace on the PCB will have a characteristic impedance, calculated with respect to its reference plane. Altium Designer has impedance calculation tools built-in; however, for high speed signals, there are many other tools to assist with simulating performance and verifying calculations.

The required trace width and dielectric spacing may be easily calculated for required impedance according to the PCB stack-up. At the same time, traces, serpentines, and pairs are better if they are separated as much as possible to prevent any crosstalk and to increase their glitch immunity.

Also, the use of stubs should be avoided. Finally, to prevent crosstalk between adjacent layers, any parallel signal routing along layers should be avoided unless there is a solid plane between them.

 
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Skew selection affinity designer free. Cropping and straightening

 
› free-transform-with-affinity-photo. To free transform with Affinity Photo, use the Move Tool (keyboard shortcut: V) for moving, scaling, rotating, and shearing objects. Use the Perspective Tool to.

 
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I have compiled a set of instructions to help you change them. With a simple design, these folder icons are the best for those who want something different from what they have and maintain it simple and practical. The Aurora folders icons are for people who want a little more. They are beautifully colored and offer a fresh new look to the folders.

The Kaori icons are a set that brings the beauty of the orient to your computer. This set is a simple pack with folders and drivers icons that are flat but give off a sense of depth with its shadows that bring out the pictures in it. Are you a Star Trek fan? Then this one is for you. Not a fan? You should still check it out. It has a simple, but different design and has a nice feeling to it. The Red Little Shoes icons is a set that looks like it was handmade. In Pixilated says it all.

Would you like to see those little classic pixels on your pc? Then this one is the right one for you. The Um Icons are a rather big icon pack with a lot of icons to choose from. Some convey the feeling of drawings, others are more realistic, and others can be a little scarier. This one is another great pack with icons that look like sketches.

It has a more childlike feeling than the previous one, but it has a nice, happy feeling to it, that just makes your day a little brighter. Just like the name say, this pack is full of flat icons with a nostalgic feeling that makes your PC look a little more retro. Silver-colored folders might not exactly be what you were looking for, but when you add the touch of a 3D pen, brush or any other realistic object, these icons may end up on your top five picks.

With beautiful and colorful images and a feeling similar to Mac, these can be great if you want to add a refreshing tone to your Windows. Do you like blue and want to add a simple and flat design to your computer? A minimalistic pack with great and simple but not always flat icons that are enjoyable for a change of style.

As a matter of fact, you could compare an icon DLL file with a library of icons. When you want to change the icon of a particular file, folder or shortcut, you can just set its icon location to point to a specific DLL file, and a specific icon from it.

Most icons from Windows 10 are stored inside DLL files. In the next section, we show you where these DLLs with icons are found in Windows We tried to organize our list of default Windows 10 icons, both by their DLL files location and by taking into consideration the primary purpose of the icons found inside the DLL files. Next, you can see our list of locations from Windows 10 where you find.

DLL files containing icons:. The imageres. It has icons for different types of folders, hardware devices, peripherals, actions, and so on. The shell Together with imageres. Inside it, you can find icons for internet, devices, networks, peripherals, folders and so on. In it, you can find funny icons that depict things such as a window, a trumpet, a ball, a wizard hat, and others.

The first icon is the default icon used by File Explorer , but the next ones were used in older Windows versions for things like printers, folders, actions, messenger apps, mail, and others. It has icons with people, mouse cursors, a magnifying glass, a mouse and a keyboard, and so on.

Among others, it includes icons for networks, folders, authentication, time, computers, and servers. For instance, inside it, you can find icons for Bluetooth, wireless routers, and network connections.

While each of the options listed above provide alternative ways to find the Downloads folder, you may be looking for a specific file that you have downloaded instead. If so, then you will need to open that browser, open its Settings menu, then find the appropriate section that specifies downloads.

In the case of Microsoft Edge, this is found by:. Most Web browsers also provide a way to access your recent downloads from that browser as well.

For example, this article will show you how to view recent downloads in Google Chrome. Matthew Burleigh has been writing tech tutorials since His writing has appeared on dozens of different websites and been read over 50 million times. However, he now works full time writing content online and creating websites.

 
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Download icon folder windows 10

 

Of course, the installation is simple, and you can change the individual folder icons as per your needs. Insignia Icon Theme is an amazing choice if you need a different approach towards file icons on Windows 10 system. Numix Circle could be one of the best Windows 10 icon packs out there if you are looking for a circular icon set. The beauty of the icon pack is in how it transforms your taskbar and desktop into a modern one in just no time.

If you want some extra shadow-ish icons for the Windows 10 interface, you should consider the Shadow icon pack for sure.

Packed with icons for almost every purpose, this icon pack would make things easier for the aesthetic enthusiast in you. While it does not have many app-based icons, it will replace almost all the Folder and Drive icons. If you are a Star Trek fan and want to add some tone of the same into your desktop and UI, you should check out this wonderful icon pack.

The Star Trek Folders icon pack is very easy to install and offers some of the best looks you can give for Windows Agua is actually an icon pack that has originally been made for macOS versions, but you would love to have the same icons on your Windows 10 task bar. The icons are known for their simple design and the overall appeal to the eyes. The problem here is that the collection is very limited, but Agua can help you when you have time to customize.

Pixelated Icon Set is your gateway if you want to give that retro look to your desktop and task bar. The icons may cause some problem to the eyes, but you will get used to the style in no time. Once everything is done and dusted, do not mind if the desktop looks like an old video game from 80s or 90s. What do you think of these icon packs? We believe they add some flare and individuality to the Windows 10 experience you want to have. We hope this list helps.

Thank you Tushar for sharing this information and the icon packs. Would love to try one of these packs. Good to know we can get these beautiful icons on the Windows too. Thanks a lot for this icon packs. But is there anything I can do about the size of these icons? They are pretty big on my screen. Great list. Loved these icons. Still confused which one to choose. Do you have anything favorite that I can try? Your words are your own, so be nice and helpful if you can. Using business name or keywords instead of your real name will lead to the comment being deleted.

Anonymous commenting is not allowed either. Limit the amount of links submitted in your comment. Comments Insignia Icon pack looks interesting. Downloading the same. Thanks for recommendation. Thanks a lot. Nice list. Do Mac users have privilege of changing icons?

Do you have anything similar to the macos icons? Copy this link in your website: Icons made by Freepik from flaticon. Don’t you want to attribute the author? Colors Display Shapes. Select a color from the icon Choose a new color.

You have exceeded the number of color samples for this palette. Save Cancel. Move left. Move right. Move up. Move down. Flip Flip horizontal. Flip vertical. Select a shape None. Rounded square. You have reached the icons limit per collection icons. Pinterest Facebook Twitter. Icons Stickers Animated icons Group by pack. Sort by Popular Popular Recent. Popular Recent. Add to collection. Copy PNG to clipboard. Download PNG. Previous page Next page. Premium download Over 7,, icons for 7.

Free download You must attribute the author Using it for the web? Copy this link in your website: Copy Copied! How likely are you to recommend Flaticon to a friend? Not likely Very likely. No thanks.

 
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