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Topic:Sarcoids

Sarcoids are a type of skin tumor commonly found in horses, characterized by their locally aggressive behavior but non-metastatic nature. These tumors are associated with bovine papillomavirus infection and can appear in various forms, including nodular, verrucous, and fibroblastic types. Sarcoids are the most frequently diagnosed equine skin tumors and can affect horses of any age, breed, or sex. They often occur on the head, limbs, and ventral abdomen, causing discomfort and potential interference with tack and equipment. This page compiles peer-reviewed research studies and scholarly articles that investigate the etiology, pathology, clinical presentation, and management strategies for equine sarcoids.
A pilot phase II study of the efficacy and biosafety of doxorubicin chemotherapy in tumor-bearing equidae.
Journal of veterinary internal medicine    September 20, 2013   Volume 27, Issue 6 1581-1588 doi: 10.1111/jvim.12144
Théon AP, Pusterla N, Magdesian KG, Wittenburg L, Marmulak T, Wilson WD.The efficacy and biosafety of a previously established tolerable dosage of doxorubicin have not been established in horses. Objective: To provide preliminary evidence of the efficacy of doxorubicin in tumor-bearing horses, explore drug pharmacokinetics profile, and estimate period of risk of exposure to drug residues. Methods: Twelve horses with 37 tumors. Methods: Treatment protocol included 6 treatments at 3-week intervals. Eight horses were uniformly treated at a dosage of 70 mg/m(2) and 4 horses received 4 of 6 treatment cycles at 70 mg/m(2) . Clinical signs, tumor responses, and toxicoses...
Surgical Management of Penile and Preputial Neoplasms in Equine with Special Reference to Partial Phallectomy.
Journal of veterinary medicine    September 8, 2013   Volume 2013 891413 doi: 10.1155/2013/891413
Rizk A, Mosbah E, Karrouf G, Abou Alsoud M.Penile and preputial neoplasia in horses occurs infrequently and represents diagnostic and therapeutic challenges. The present study was carried out on a total number of 21 equids (14 stallions and 7 donkeys) suffered from different penile and preputial neoplasia. Diagnosis of neoplasms was based up on history of the case, clinical examination as well as histopathological evaluation. Animals with penile and preputial neoplasms were underwent local excision and partial phallectomy with a slightly modified version of the techniques described by William's. The diagnosed neoplasms were penile and ...
Papillomavirus associated diseases of the horse.
Veterinary microbiology    August 18, 2013   Volume 167, Issue 1-2 159-167 doi: 10.1016/j.vetmic.2013.08.003
Nasir L, Brandt S.The Papillomaviridae family comprises a large number of viruses that can infect a broad range of hosts including mammals, birds and reptiles giving rise to benign lesions of the skin or mucosal membranes. They are characterized by great genetic diversity yet adhere to common biological principles. In this review, we first describe the structure and function of the viral proteins encoded by papillomaviruses (PVs), with a particular emphasis on bovine papillomaviruses (BPV). We then discuss the role of BPV types 1 and 2 in the pathogenesis of equine sarcoids and present recent evidence implicati...
Design of NK-2-derived peptides with improved activity against equine sarcoid cells.
Journal of peptide science : an official publication of the European Peptide Society    July 25, 2013   Volume 19, Issue 10 619-628 doi: 10.1002/psc.2540
Gross S, Wilms D, Krause J, Brezesinski G, Andrä J.Equine sarcoid is a topically accessible model for the evaluation of anticancer peptides acting by physical membrane disruption avoiding the complexity of a systemic application. We aim at evaluating and improving natural peptides for host defence as lead structures, where we focus on the cationic and amphipathic peptide NK-2. Cytotoxicity tests, fluorescence microscopy and a chip-based biosensor, which enabled real-time monitoring of cell metabolism, were applied. Cancer cell killing was dynamic with an initial phase of increased cellular respiration, followed by membrane destruction. NK-2 wa...
BAG3 protects bovine papillomavirus type 1-transformed equine fibroblasts against pro-death signals.
Veterinary research    July 22, 2013   Volume 44, Issue 1 61 doi: 10.1186/1297-9716-44-61
Cotugno R, Gallotta D, d'Avenia M, Corteggio A, Altamura G, Roperto F, Belisario MA, Borzacchiello G.In human cancer cells, BAG3 protein is known to sustain cell survival. Here, for the first time, we demonstrate the expression of BAG3 protein both in equine sarcoids in vivo and in EqS04b cells, a sarcoid-derived fully transformed cell line harbouring bovine papilloma virus (BPV)-1 genome. Evidence of a possible involvement of BAG3 in equine sarcoid carcinogenesis was obtained by immunohistochemistry analysis of tumour samples. We found that most tumour samples stained positive for BAG3, even though to a different grade, while normal dermal fibroblasts from healthy horses displayed very weak ...
Analysis of activated platelet-derived growth factor β receptor and Ras-MAP kinase pathway in equine sarcoid fibroblasts.
BioMed research international    July 11, 2013   Volume 2013 283985 doi: 10.1155/2013/283985
Altamura G, Corteggio A, Nasir L, Yuan ZQ, Roperto F, Borzacchiello G.Equine sarcoids are skin tumours of fibroblastic origin affecting equids worldwide. Bovine papillomavirus type-1 (BPV-1) and, less commonly, type-2 are recognized as etiological factors of sarcoids. The transforming activity of BPV is related to the functions of its major oncoprotein E5 which binds to the platelet-derived growth factor β receptor (PDGFβR) causing its phosphorylation and activation. In this study, we demonstrate, by coimmunoprecipitation and immunoblotting, that in equine sarcoid derived cell lines PDGFβR is phosphorylated and binds downstream molecules related to Ras-mitoge...
Bovine papillomavirus type 13 DNA in equine sarcoids.
Journal of clinical microbiology    May 1, 2013   Volume 51, Issue 7 2167-2171 doi: 10.1128/JCM.00371-13
Lunardi M, de Alcântara BK, Otonel RA, Rodrigues WB, Alfieri AF, Alfieri AA.Equine sarcoids are locally aggressive fibroblastic neoplasms considered to be the most common skin tumors of horses worldwide. Bovine papillomavirus types 1 and 2 have typically been associated with sarcoids in equids. Investigations aiming to identify papillomavirus strains, aside from bovine papillomaviruses 1 and 2, which might be associated with sarcoid lesions, have been lacking. The aim of this article is to report the identification of a third bovine papillomavirus type, bovine papillomavirus 13, associated with equine sarcoids. Six sarcoid lesions were collected from diverse anatomica...
Equine sarcoidosis: clinical signs, diagnosis, treatment and outcome of 22 cases.
Veterinary dermatology    January 22, 2013   Volume 24, Issue 1 218-24.e48 doi: 10.1111/j.1365-3164.2012.01108.x
Sloet van Oldruitenborgh-Oosterbaan MM, Grinwis GC.Equine sarcoidosis may present as a generalized or localized exfoliative dermatitis and/or as a granulomatous inflammation of multiple organs. Objective: To report the clinical signs, diagnosis, treatment and outcome for 22 horses with histologically confirmed sarcoidosis. Methods: Twenty-two horses of different breeds, between 3 and 17 years of age. Methods: Diagnosis was based on clinical signs and histopathological findings. Results: The following three forms of equine sarcoidosis were identified: generalized (13.6%), partially generalized (18.2%) and localized (68.2%). High-dose systemic c...
Cutaneous neoplastic lesions of equids in the central United States and Canada: 3,351 biopsy specimens from 3,272 equids (2000-2010).
Journal of the American Veterinary Medical Association    December 14, 2012   Volume 242, Issue 1 99-104 doi: 10.2460/javma.242.1.99
Schaffer PA, Wobeser B, Martin LE, Dennis MM, Duncan CG.To identify epidemiological trends in cutaneous neoplasms affecting equids in central North America and compare them with previously reported trends. Methods: Retrospective case series. Methods: 3,351 cutaneous biopsy specimens from 3,272 equids with a neoplastic diagnosis. Methods: Diagnostic reports from 2 diagnostic laboratories (Colorado State University and Prairie Diagnostic Services Inc) were reviewed for frequency of specific lesions and epidemiological trends. Variables included in analyses (if known) were age, sex, breed, geographic location, date of diagnosis, location of neoplasm o...
Equine sarcoids: Bovine Papillomavirus type 1 transformed fibroblasts are sensitive to cisplatin and UVB induced apoptosis and show aberrant expression of p53.
Veterinary research    December 4, 2012   Volume 43, Issue 1 81 doi: 10.1186/1297-9716-43-81
Finlay M, Yuan Z, Morgan IM, Campo MS, Nasir L.Bovine papillomavirus type 1 infects not only cattle but also equids and is a causative factor in the pathogenesis of commonly occurring equine sarcoid tumours. Whilst treatment of sarcoids is notoriously difficult, cisplatin has been shown to be one of the most effective treatment strategies for sarcoids. In this study we show that in equine fibroblasts, BPV-1 sensitises cells to cisplatin-induced and UVB-induced apoptosis, a known cofactor for papillomavirus associated disease, however BPV-1 transformed fibroblasts show increased clonogenic survival, which may potentially limit the therapeut...
O(6)-methylguanine-DNA methyltransferase in equine sarcoids: molecular and epigenetic analysis.
BMC veterinary research    November 10, 2012   Volume 8 218 doi: 10.1186/1746-6148-8-218
Altamura G, Strazzullo M, Corteggio A, Francioso R, Roperto F, D'Esposito M, Borzacchiello G.Bovine papillomaviruses (BPVs) types 1 and 2 are the only known papillomaviruses able to jump the species. In fact, BPVs 1/2 induce neoplasia in their natural bovine host but infection is also associated to neoplastic skin lesions in equids termed sarcoids. The equine sarcoid is considered to be the most common equine cutaneous tumour worldwide for which no effective therapy is available. Very little is known about the molecular mechanisms underlying tumourigenesis, although genes contributing to sarcoid development have been identified. Several studies associate the development of cancer to t...
[Successful treatment of an equine sarcoid. Case report on a combined surgical and photodynamic therapy].
Tierarztliche Praxis. Ausgabe G, Grosstiere/Nutztiere    October 19, 2012   Volume 40, Issue 5 309-313 
Reschke C.The case report describes the surgical and photodynamic treatment (PDT) of an equine sarcoid in a 6-year-old gelding. A mass on the ventral prepuce, several tumours on the lateral aspect of the prepuce and one sarcoid on the front aspect of the chest were treated. For PDT, Temoporfin (Fospeg® Biolitec AG, Jena) at a concentration of 0.15mg/ml was injected locally. The subsequent irradiation was performed using a red-light laser (652nm) with an energy density of 10J/cm². The mass on the ventral aspect of the prepuce and some of the lateral tumours displayed total remission. The remaining tumo...
Characterisation of early and late bovine papillomavirus protein expression in equine sarcoids.
Veterinary microbiology    October 17, 2012   Volume 162, Issue 2-4 369-380 doi: 10.1016/j.vetmic.2012.10.010
Wilson AD, Armstrong ELR, Gofton RG, Mason J, De Toit N, Day MJ.Sarcoids are common skin tumours of horses and donkeys that are characterised by persistent proliferation of dermal fibroblasts associated with the presence of bovine papillomavirus (BPV) DNA. Some early BPV proteins have been demonstrated within sarcoids and RNA containing both early and late transcripts is present, yet it remains unclear whether late replication of BPV, culminating in the production of infectious virus particles, can occur in equids. Here we report that BPV1 RNA isolated from equine sarcoids encodes a unique deletion of four residues within the L2 protein suggesting a novel ...
Treatment of equine sarcoids.
The Veterinary record    October 2, 2012   Volume 171, Issue 13 330 doi: 10.1136/vr.e6506
Scagliarini A, Bettini G, Savini F, Spadari A, Sonvico F, Balducci AG, Calbucci S, Colombo G.No abstract available
DNA hypomethylation and oxidative stress-mediated increase in genomic instability in equine sarcoid-derived fibroblasts.
Biochimie    May 29, 2012   Volume 94, Issue 9 2013-2024 doi: 10.1016/j.biochi.2012.05.026
Potocki L, Lewinska A, Klukowska-Rötzler J, Bugno-Poniewierska M, Koch C, Mählmann K, Janda J, Wnuk M.It is widely accepted that equine sarcoid disease, the most common skin associated neoplasm in equids, is induced by bovine papillomavirus (BPV-1). Although BPV-1 DNA has been found in almost all examined sarcoids so far, its detailed impact on the horse's host cell metabolism is largely unknown. We used equine fibroblast cell lines originating from sarcoid biopsies to study BPV-1-associated changes on DNA methylation status and oxidative stress parameters. Sarcoid-derived fibroblasts manifested increased proliferation in vitro, transcriptional rDNA activity (NORs expression) and DNA hypometh...
Mycobacterium DNA detection in liver and skin of a horse with generalized sarcoidosis. Oliveira-Filho JP, Monteiro LN, Delfiol DJ, Sequeira JL, Amorim RM, Fabris VE, Del Piero F, Borges AS.Sarcoidosis is a rare equine skin disease characterized primarily by an exfoliative and granulomatous dermatitis but also presenting granulomatous inflammation of multiple systems. The current report presents the clinical and histopathological findings of sarcoidosis in a 16-year-old American Quarter Horse gelding with nested polymerase chain reaction Mycobacterium spp. DNA detection within hepatic and skin samples. Mycobacterium spp. may play a role in the pathogenesis of equine sarcoidosis as has been proposed for human sarcoidosis.
Molecular and epigenetic analysis of the fragile histidine triad tumour suppressor gene in equine sarcoids.
BMC veterinary research    March 16, 2012   Volume 8 30 doi: 10.1186/1746-6148-8-30
Strazzullo M, Corteggio A, Altamura G, Francioso R, Roperto F, D'Esposito M, Borzacchiello G.Sarcoids are peculiar equine benign tumours. Their onset is associated with Bovine Papillomavirus type -1 or -2 (BPV-1/2) infection. Little is known about the molecular interplay between viral infection and neoplastic transformation. The data regarding papillomavirus infections in human species show the inactivation of a number of tumour suppressor genes as basic mechanism of transformation. In this study the putative role of the tumour suppressor gene Fragile Histidine Triad (FHIT) in sarcoid tumour was investigated in different experimental models. The expression of the oncosuppressor protei...
The approach to the equine dermatology case in practice.
The Veterinary clinics of North America. Equine practice    February 28, 2012   Volume 28, Issue 1 131-153 doi: 10.1016/j.cveq.2012.01.004
Knottenbelt DC.A logical and thorough clinical investigation should provide the best basis for the diagnosis of skin diseases. Where no diagnosis can be reached despite a full range of investigations, the clinician can justifiably attempt symptomatic treatment, but it is always better to focus treatment on a specific condition based on properly accumulated and tested clinical evidence. Unfortunately, in equine dermatology there are few text descriptions of the majority of the conditions encountered in practice. While a few diseases are well recognized, there is still little consensus on the best treatments f...
Whole genome scan identifies several chromosomal regions linked to equine sarcoids.
Schweizer Archiv fur Tierheilkunde    January 10, 2012   Volume 154, Issue 1 19-25 doi: 10.1024/0036-7281/a000288
Jandova V, Klukowska-Rötzler J, Dolf G, Janda J, Roosje P, Marti E, Koch C, Gerber V, Swinburne J.Despite the evidence for a genetic predisposition to develop equine sarcoids (ES), no whole genome scan for ES has been performed to date. The objective of this explorative study was to identify chromosome regions associated with ES. The studied population was comprised of two half-sibling sire families, involving a total of 222 horses. Twenty-six of these horses were affected with ES. All horses had been previously genotyped with 315 microsatellite markers. Quantitative trait locus (QTL) signals were suggested where the F statistic exceeded chromosome-wide significance at P < 0.05. The QTL...
Localization of Bovine papillomavirus in equine sarcoids and inflammatory skin conditions of horses using laser microdissection and two forms of DNA amplification. Wobeser BK, Hill JE, Jackson ML, Kidney BA, Mayer MN, Townsend HG, Allen AL.Equine sarcoids are the most common tumor of horses. Bovine papillomavirus (BPV) has been suggested as the cause of sarcoids. Studies have shown that BPV is present in swabs or biopsies from nonsarcoid-bearing equine skin. Skin biopsies from a variety of different conditions and normal skin from horses with no reported history of sarcoids were examined by polymerase chain reaction (PCR) for the presence of BPV, which was found in all different types of skin conditions as well as normal skin. Forty-one out of 86 skin biopsies from horses without sarcoids were found to contain BPV DNA. Laser mic...
Treatment of equine sarcoid in seven Cape mountain zebra (Equus zebra zebra).
Journal of wildlife diseases    November 22, 2011   Volume 47, Issue 4 917-924 doi: 10.7589/0090-3558-47.4.917
Marais HJ, Page PC.Equine sarcoid has been diagnosed in endangered Cape mountain zebra (Equus zebra zebra) in at least two game reserves in South Africa, with prevalence as high as 53% in Bontebok National Park. Seven Cape mountain zebras with sarcoids were treated with either surgical excision, 5-fluorouracil, allogenous vaccine, or a combination of 5-fluorouracil and allogenous vaccine. One of the two sarcoids on one of the 5-fluorouracil-treated zebras was left untreated. The microscopic features of the tumors evaluated showed either all or most of the typical epidermal and dermal histologic features of equin...
Safety and immunogenicity of BPV-1 L1 virus-like particles in a dose-escalation vaccination trial in horses.
Equine veterinary journal    September 6, 2011   Volume 44, Issue 1 107-111 doi: 10.1111/j.2042-3306.2011.00390.x
Hainisch EK, Brandt S, Shafti-Keramat S, Van den Hoven R, Kirnbauer R.Infection with bovine papillomaviruses types 1 and 2 (BPV-1, BPV-2) can lead to the development of therapy-resistant skin tumours termed sarcoids and possibly other skin diseases in equids. Although sarcoids seriously compromise the welfare of affected animals and cause considerable economic losses, no prophylactic vaccine is available to prevent this common disease. In several animal species and man, immunisation with papillomavirus-like particles (VLP) has been shown to protect efficiently from papillomaviral infection. Objective: BPV-1 L1 VLPs may constitute a safe and highly immunogenic va...
Successful treatment of equine sarcoids with cisplatin electrochemotherapy: a retrospective study of 48 cases.
Equine veterinary journal    July 27, 2011   Volume 44, Issue 2 214-220 doi: 10.1111/j.2042-3306.2011.00425.x
Tamzali Y, Borde L, Rols MP, Golzio M, Lyazrhi F, Teissie J.Sarcoids are the commonest form of equine skin tumour. Several therapeutic measures have been described but none is considered to be universally effective. Electrochemotherapy (ECT) is a new anticancer therapy that utilises electrical field pulses to induce increased cell membrane permeability to antitumour hydrophilic drugs, such as cisplatin. The increased intracellular concentration of the drugs has a significant therapeutic benefit. The procedure has not been previously reported in a large number of horses. Objective: To validate ECT as a novel alternative treatment for equine sarcoids. Me...
Upregulation of equine matrix metalloproteinase 1 by bovine papillomavirus type 1 is through the transcription factor activator protein-1.
The Journal of general virology    July 20, 2011   Volume 92, Issue Pt 11 2608-2619 doi: 10.1099/vir.0.033431-0
Yuan Z, Gault EA, Campo MS, Nasir L.Equine sarcoids represent the most common skin tumours in equids worldwide, characterized by extensive invasion and infiltration of lymphatics, rare regression and high recurrence after surgical intervention. Bovine papillomavirus type 1 (BPV-1) activity is necessary for the transformation phenotype of equine fibroblasts. Among the many changes induced by BPV-1, matrix metalloproteinase 1 (MMP-1) upregulation contributes to the invasiveness of equine fibroblasts. However, it is not yet known how BPV-1 proteins regulate equine MMP-1 expression. To elucidate this mechanism, the equine MMP-1 prom...
Inoculation of young horses with bovine papillomavirus type 1 virions leads to early infection of PBMCs prior to pseudo-sarcoid formation.
The Journal of general virology    June 29, 2011   Volume 92, Issue Pt 10 2437-2445 doi: 10.1099/vir.0.033670-0
Hartl B, Hainisch EK, Shafti-Keramat S, Kirnbauer R, Corteggio A, Borzacchiello G, Tober R, Kainzbauer C, Pratscher B, Brandt S.Bovine papillomavirus types 1 and 2 (BPV-1 and BPV-2) are known to induce common equine skin tumours, termed sarcoids. Recently, it was demonstrated that vaccination with BPV-1 virus-like particles (VLPs) is safe and highly immunogenic in horses. To establish a BPV-1 challenge model for evaluation of the protective potential of BPV-1 VLPs, four foals were injected intradermally with infectious BPV-1 virions and with viral genome-based and control inocula, and monitored daily for tumour development. Blood was taken before inoculation and at weekly intervals. BPV-1-specific serum antibodies were...
p38 mitogen-activated protein kinase is crucial for bovine papillomavirus type-1 transformation of equine fibroblasts.
The Journal of general virology    April 6, 2011   Volume 92, Issue Pt 8 1778-1786 doi: 10.1099/vir.0.031526-0
Yuan Z, Gault EA, Campo MS, Nasir L.Equine sarcoids represent the most common skin tumours in equids worldwide, characterized by extensive invasion and infiltration of lymphatics, rare regression and high recurrence after surgical intervention. Bovine papillomavirus type-1 (BPV-1) and less commonly BPV-2 are the causative agents of the diseases. It has been demonstrated that BPV-1 viral gene expression is necessary for maintaining the transformation phenotype. However, the underlying mechanism for BPV-1 transformation remains largely unknown, and the cellular factors involved in transformation are not fully understood. Previousl...
Successful treatment of equine sarcoids by topical aciclovir application.
The Veterinary record    February 11, 2011   Volume 168, Issue 7 187 doi: 10.1136/vr.c5430
Stadler S, Kainzbauer C, Haralambus R, Brehm W, Hainisch E, Brandt S.Based on the anecdotally reported eradication of a sarcoid using aciclovir cream, the curative potential of this ointment was investigated in 22 sarcoid-affected horses referred to the Equine Clinic Tillysburg, Austria, between 2006 and 2009. Sarcoid disease was diagnosed by clinical examination and bovine papillomavirus types 1 and 2 from intact skin and tumour tissue. As nine horses had more than one lesion, a total of 47 sarcoids were treated by daily topical application of aciclovir 5 per cent cream for a period of two to six months; in four horses, surgical tumour ablation was performed b...
Epidemiology of equine sarcoids in horses in western Canada.
The Canadian veterinary journal = La revue veterinaire canadienne    January 5, 2011   Volume 51, Issue 10 1103-1108 
Wobeser BK, Davies JL, Hill JE, Jackson ML, Kidney BA, Mayer MN, Townsend HG, Allen AL.Sarcoids are the most common tumor of the equine skin but only 1 study describing the epidemiology of sarcoids in Canadian horses has been published. The records of 5 veterinary diagnostic laboratories in western Canada were searched to identify submissions of sarcoids from horses. The submission records and diagnostic reports of 802 separate submissions of equine sarcoids were reviewed for age, breed, and gender of the horse and the number, location, and clinical type of sarcoid. From these records, the 307 submissions to laboratories in Saskatchewan were compared to a reference group to test...
Phylogenetic analysis of bovine papillomavirus E5 detected in equine sarcoids in Poland.
Polish journal of veterinary sciences    January 1, 2011   Volume 14, Issue 4 653-654 doi: 10.2478/v10181-011-0096-6
Szczerba-Turek A, Siemionek J, Bancerz-Kisiel A, Raś A, Szweda W.The aim of the study was to analyse a part of the sequence of the E5 gene of bovine papillomaviruses (BPV) associated with equine sarcoids in Polish horses. Samples of 40 skin lesions obtained from 29 horses were collected for molecular examination. The PCR amplicons of BPV DNA were detected in 38 specimens. After phylogenetic analysis 37 specimens were recognized as BPV-1 and one as BPV-2. Phylogenetic analysis has allowed the classification of the amplicons into two phylogenetic groups (A1,) and four separate isolates (2, 10, 16, 17).
BPV-1 infection is not confined to the dermis but also involves the epidermis of equine sarcoids.
Veterinary microbiology    December 23, 2010   Volume 150, Issue 1-2 35-40 doi: 10.1016/j.vetmic.2010.12.021
Brandt S, Tober R, Corteggio A, Burger S, Sabitzer S, Walter I, Kainzbauer C, Steinborn R, Nasir L, Borzacchiello G.In equids, bovine papillomaviruses of type 1 (BPV-1) and less frequently type 2 induce common, locally aggressive skin tumours termed sarcoids. Whereas BPV infection in cattle usually involves the epidermis and is productive in this skin layer, infection in equids is currently thought to be abortive, with virus solely residing as multiple episomes in dermal fibroblasts. Based on recent observations that do not agree with this assumption, we hypothesised that BPV also infects equid epidermis and is active in this skin layer. To test this hypothesis, we conducted a proof-of-principle study on ei...
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