COMPARISON BETWEEN PORTALS FOR PLACEMENT OF ANCHORS IN SHOULDER INSTABILITY.

Objective: to radiographically compare the effects of anchor positioning in the arthroscopic treatment of shoulder instability, in the 3- and 5-o'clock portals. Methods: retrospective study of 36 patients, operated by two shoulder surgeons at the Unimed BH hospital, between January 2013 and January 2018. Each surgeon used only one of either the 3- or the 5-o'clock portal. After postoperative radiographs we performed angle comparisons between the greatest glenoidal axis, the angle of anchor insertion and distance from the inferior pole. Results: the 5-o'clock portal provided better placement than its 3-o'clock counterpart, which allowed for greater orthogonality in relation to the glenoid rim (p < 0.05). Conclusion: the 5-o'clock portal allowed for better anchor placement than the 3 o'clock one. Level of Evidence II, Clinical Trial.

Objetivo: Comparar radiograficamente o posicionamento das âncoras utilizadas no tratamento artroscópico da instabilidade do ombro, através dos portais de 3 ou 5 horas. Métodos: Avaliação retrospectivae de 36 pacientes, operados por dois cirurgiões de ombro do Hospital Unimed BH, entre janeiro de 2013 e janeiro de 2018. Cada cirurgião utilizou apenas uma das técnicas ­ portal de 3 ou 5 horas. As radiografias pós-operatórias foram avaliadas e comparadas a angulações entre o maior eixo da glenoide, o ângulo de inserção da âncora e a distância em relação ao polo inferior. Resultados: A utilização do portal de 5 horas propiciou a colocação mais adequada das âncoras em relação ao portal de 3 horas, permitindo o posicionamento mais ortogonal em relação à borda da glenoide (p < 0,05). Conclusão: A utilização do portal de 5 horas apresenta melhor posicionamento das âncoras quando comparado ao portal de 3 horas. Nível de evidência II, Ensaio Clínico.

Contouring aid tools in radiotherapy. Smoothing: the false friend.

OBJECTIVE: Contouring accuracy is critical in modern radiotherapy. Several tools are available to assist clinicians in this task. This study aims to evaluate the performance of the smoothing tool in the ARIA system to obtain more consistent volumes. METHODS: Eleven different geometric shapes were delineated in ARIA v15.6 (Sphere, Cube, Square Prism, Six-Pointed Star Prism, Arrow Prism, And Cylinder and the respective volumes at 45° of axis deviation (_45)) in 1, 3, 5, 7, and 10 cm side or diameter each. Post-processing drawing tools to smooth those first-generated volumes were applied in different options (2D-ALL vs 3D) and grades (1, 3, 5, 10, 15, and 20). These volumetric transformations were analyzed by comparing different parameters: volume changes, center of mass, and DICE similarity coefficient index. Then we studied how smoothing affected two different volumes in a head and neck cancer patient: a single rounded node and the volume delineating cervical nodal areas. RESULTS: No changes in data were found between 2D-ALL or 3D smoothing. Minimum deviations were found (range from 0 to 0.45 cm) in the center of mass. Volumes and the DICE index decreased as the degree of smoothing increased. Some discrepancies were found, especially in figures with cleft and spikes that behave differently. In the clinical case, smoothing should be applied only once throughout the target delineation process, preferably in the largest volume (PTV) to minimize errors. CONCLUSION: Smoothing is a good tool to reduce artifacts due to the manual delineation of radiotherapy volumes. The resulting volumes must be always carefully reviewed.

MET receptor serves as a promising target in melanoma brain metastases.

The development of brain metastases hallmarks disease progression in 20-40% of melanoma patients and is a serious obstacle to therapy. Understanding the processes involved in the development and maintenance of melanoma brain metastases (MBM) is critical for the discovery of novel therapeutic strategies. Here, we generated transcriptome and methylome profiles of MBM showing high or low abundance of infiltrated Iba1high tumor-associated microglia and macrophages (TAMs). Our survey identified potential prognostic markers of favorable disease course and response to immune checkpoint inhibitor (ICi) therapy, among them APBB1IP and the interferon-responsive gene ITGB7. In MBM with high ITGB7/APBB1IP levels, the accumulation of TAMs correlated significantly with the immune score. Signature-based deconvolution of MBM via single sample GSEA revealed enrichment of interferon-response and immune signatures and revealed inflammation, stress and MET receptor signaling. MET receptor phosphorylation/activation maybe elicited by inflammatory processes in brain metastatic melanoma cells via stroma cell-released HGF. We found phospho-METY1234/1235 in a subset of MBM and observed a marked response of brain metastasis-derived cell lines (BMCs) that lacked druggable BRAF mutations or developed resistance to BRAF inhibitors (BRAFi) in vivo to MET inhibitors PHA-665752 and ARQ197 (tivantinib). In summary, the activation of MET receptor in brain colonizing melanoma cells by stromal cell-released HGF may promote tumor self-maintenance and expansion and might counteract ICi therapy. Therefore, therapeutic targeting of MET possibly serves as a promising strategy to control intracranial progressive disease and improve patient survival.

An atlas of cells in the human tonsil.

Palatine tonsils are secondary lymphoid organs (SLOs) representing the first line of immunological defense against inhaled or ingested pathogens. We generated an atlas of the human tonsil composed of >556,000 cells profiled across five different data modalities, including single-cell transcriptome, epigenome, proteome, and immune repertoire sequencing, as well as spatial transcriptomics. This census identified 121 cell types and states, defined developmental trajectories, and enabled an understanding of the functional units of the tonsil. Exemplarily, we stratified myeloid slan-like subtypes, established a BCL6 enhancer as locally active in follicle-associated T and B cells, and identified SIX5 as putative transcriptional regulator of plasma cell maturation. Analyses of a validation cohort confirmed the presence, annotation, and markers of tonsillar cell types and provided evidence of age-related compositional shifts. We demonstrate the value of this resource by annotating cells from B cell-derived mantle cell lymphomas, linking transcriptional heterogeneity to normal B cell differentiation states of the human tonsil.

Comparison between portals for placement of anchors in shoulder instability / Comparação entre portais para colocação de âncoras na instabilidade do ombro

ABSTRACT Objective: to radiographically compare the effects of anchor positioning in the arthroscopic treatment of shoulder instability, in the 3- and 5-o'clock portals. Methods: retrospective study of 36 patients, operated by two shoulder surgeons at the Unimed BH hospital, between January 2013 and January 2018. Each surgeon used only one of either the 3- or the 5-o'clock portal. After postoperative radiographs we performed angle comparisons between the greatest glenoidal axis, the angle of anchor insertion and distance from the inferior pole. Results: the 5-o'clock portal provided better placement than its 3-o'clock counterpart, which allowed for greater orthogonality in relation to the glenoid rim (p < 0.05). Conclusion: the 5-o'clock portal allowed for better anchor placement than the 3 o'clock one. Level of Evidence II, Clinical Trial.

RESUMO Objetivo: Comparar radiograficamente o posicionamento das âncoras utilizadas no tratamento artroscópico da instabilidade do ombro, através dos portais de 3 ou 5 horas. Métodos: Avaliação retrospectivae de 36 pacientes, operados por dois cirurgiões de ombro do Hospital Unimed BH, entre janeiro de 2013 e janeiro de 2018. Cada cirurgião utilizou apenas uma das técnicas - portal de 3 ou 5 horas. As radiografias pós-operatórias foram avaliadas e comparadas a angulações entre o maior eixo da glenoide, o ângulo de inserção da âncora e a distância em relação ao polo inferior. Resultados: A utilização do portal de 5 horas propiciou a colocação mais adequada das âncoras em relação ao portal de 3 horas, permitindo o posicionamento mais ortogonal em relação à borda da glenoide (p < 0,05). Conclusão: A utilização do portal de 5 horas apresenta melhor posicionamento das âncoras quando comparado ao portal de 3 horas. Nível de evidência II, Ensaio Clínico.

Increasing risk of invasions by organisms on marine debris in the Southeast coast of India.

Increasing amount of anthropogenic litter in the marine environment has provided an enormous number of substrates for a wide range of marine organisms, thus serving as a potential vector for the transport of fouling organisms. Here, we examined the fouling organisms on different types of stranded litter (plastic, glass, rubber, foam sponge, cloth, metal and wood) on eight beaches along the southeast coast of India. In total, 17 encrusting species belonging to seven phyla (Arthropoda, Bryozoa, Mollusca, Annelida, Cnidaria, Chlorophyta and Foraminifera) were identified on 367 items, with one invasive species, the mussel Mytella strigata, detected. The most common species associated with marine litter were the cosmopolitan bryozoans Jellyella tuberculata (%O = 31.64 %) and J. eburnea (28.61 %), the barnacle species Lepas anserifera (29.97 %), Amphibalanus amphitrite (22.34 %) and Amphibalanus sp. (14.16 %), and the oyster species Saccostrea cucullata (13.62 %) and Magallana bilineata (5.44 %). We also reported the first records on stranded litter of four species: the gastropod species Pirenella cingulata and Umbonium vestiarium, the foraminiferan Ammonia beccarii, and the oyster M. bilineata. This study is thus the first documentation of marine litter as a vector for species dispersal in India, where the production and consumption of plastic rank among the highest in the world. We also highlight the increasing risk of invasions by non-indigenous organisms attached to debris along the southeast coast of India. Comprehensive monitoring efforts are thus needed to elucidate the type of vectors responsible for the arrival of invasive species in this region. Raising awareness and promoting education are vital components in fostering sustainable solutions to combat plastic pollution in the country and globally.

The phenotypic presentation of adult individuals with SLC6A1-related neurodevelopmental disorders.

Introduction: SLC6A1 is one of the most common monogenic causes of epilepsy and is a well-established cause of neurodevelopmental disorders. SLC6A1-neurodevelopmental disorders have a consistent phenotype of mild to severe intellectual disability (ID), epilepsy, language delay and behavioral disorders. This phenotypic description is mainly based on knowledge from the pediatric population. Method: Here, we sought to describe patients with SLC6A1 variants and age above 18 years through the ascertainment of published and unpublished patients. Unpublished patients were ascertained through international collaborations, while previously published patients were collected through a literature search. Results: A total of 15 adult patients with SLC6A1 variants were included. 9/13 patients had moderate to severe ID (data not available in two). Epilepsy was prevalent (11/15) with seizure types such as absence, myoclonic, atonic, and tonic-clonic seizures. Epilepsy was refractory in 7/11, while four patients were seizure free with lamotrigine, valproate, or lamotrigine in combination with valproate. Language development was severely impaired in five patients. Behavioral disorders were reported in and mainly consisted of autism spectrum disorders and aggressive behavior. Schizophrenia was not reported in any of the patients. Discussion: The phenotype displayed in the adult patients presented here resembled that of the pediatric cohort with ID, epilepsy, and behavioral disturbances, indicating that the phenotype of SLC6A1-NDD is consistent over time. Seizures were refractory in >60% of the patients with epilepsy, indicating the lack of targeted treatment in SLC6A1-NDDs. With increased focus on repurposing drugs and on the development of new treatments, hope is that the outlook reflected here will change over time. ID appeared to be more severe in the adult patients, albeit this might reflect a recruitment bias, where only patients seen in specialized centers were included or it might be a feature of the natural history of SLC6A1-NDDs. This issue warrants to be explored in further studies in larger cohorts.

Corrigendum: Intrafamilial variability in SLC6A1-related neurodevelopmental disorders.

[This corrects the article DOI: 10.3389/fnins.2023.1219262.].

Intrafamilial variability in SLC6A1-related neurodevelopmental disorders.

Introduction: Phenotypic spectrum of SLC6A1-related neurodevelopmental disorders (SLC6A1-NDD) includes intellectual disability (ID), autistic spectrum disorders (ASD), epilepsy, developmental delay, beginning from early infancy or after seizure onset, and other neurological features such as hypotonia and movement disorders. Data on familial phenotypic heterogeneity have been rarely reported, thus in our study we aimed to investigate intrafamilial phenotypic variability in families with SLC6A1 variants. Methods: We collected clinical, laboratory and genetic data on 39 individuals, including 17 probands, belonging to 13 families harboring inherited variants of SLC6A1. Data were collected through an international network of Epilepsy and Genetic Centers. Results: Main clinical findings in the whole cohort of 39 subjects were: (a) epilepsy, mainly presenting with generalized seizures, reported in 71% of probands and 36% of siblings or first/second-degree relatives. Within a family, the same epilepsy type (generalized or focal) was observed; (b) ID reported in 100% and in 13% of probands and siblings or first/second-degree relatives, respectively; (c) learning disabilities detected in 28% of the SLC6A1 carriers, all of them were relatives of a proband; (d) around 51% of the whole cohort presented with psychiatric symptoms or behavioral disorders, including 82% of the probands. Out of the 19 patients with psychiatric symptoms, ASD were diagnosed in 40% of them; (e) neurological findings (primarily tremor and speech difficulties) were observed 38.5% of the whole cohort, including 10 probands. Our families harbored 12 different SLC6A1 variants, one was a frameshift, two stop-gain, while the remaining were missense. No genotype-phenotype associations were identified. Discussion: Our study showed that first-or second-degree relatives presented with a less severe phenotype, featuring mainly mild intellectual and/or learning disabilities, at variance with the probands who suffered from moderate to severe ID, generalized, sometimes intractable, epileptic seizures, behavioral and psychiatric disorders. These findings may suggest that a proportion of individuals with mild SLC6A1-NDD might be missed, in particular those with an older age where genetic testing is not performed. Further studies on intrafamilial phenotypic variability are needed to confirm our results and possibly to expand the phenotypic spectrum of these disorders and benefit genetic counseling.

The dark side of artificial greening: Plastic turfs as widespread pollutants of aquatic environments.

Artificial turf (AT) is a surfacing material that simulates natural grass by using synthetic, mainly plastic, fibers in different shapes, sizes and properties. AT has spread beyond sports facilities and today shapes many urban landscapes, from private lawns to rooftops and public venues. Despite concerns regarding the impacts of AT, little is known about the release of AT fibers into natural environment. Here, for the first time, we specifically investigate the presence of AT fibers in river and ocean waters as major conduits and final destination of plastic debris transported by water runoff. Our sampling survey showed that, AT fibers - composed mainly of polyethylene and polypropylene - can constitute over 15% of the mesoplastics and macroplastics content, suggesting that AT fibers may contribute significantly to plastic pollution. Up to 20,000 fibers a day flowed down through the river, and up to 213,200 fibers per km2 were found floating on the sea surface of nearshore areas. AT, apart from impacting on urban biodiversity, urban runoff, heat island formation, and hazardous chemical leaching, is a major source of plastic pollution to natural aquatic environments.

Detection of faecal bacteria and antibiotic resistance genes in biofilms attached to plastics from human-impacted coastal areas.

Plastics have been proposed as vectors of bacteria as they act as a substrate for biofilms. In this study, we evaluated the abundance of faecal and marine bacteria and antibiotic resistance genes (ARGs) from biofilms adhered to marine plastics. Floating plastics and plastics from sediments were collected in coastal areas impacted by human faecal pollution in the northwestern Mediterranean Sea. Culture and/or molecular methods were used to quantify faecal indicators (E. coli, Enterococci and crAssphage), and the ARGs sulI, tetW and blaTEM and the 16S rRNA were detected by qPCR assays. Pseudomonas and Vibrio species and heterotrophic marine bacteria were also analysed via culture-based methods. Results showed that, plastic particles covered by bacterial biofilms, primarily consisted of marine bacteria including Vibrio spp. Some floating plastics had a low concentration of viable E. coli and Enterococci (42% and 67% of the plastics respectively). Considering the median area of the plastics, we detected an average of 68 cfu E. coli per item, while a higher concentration of E. coli was detected on individual plastic items, when compared with 100 ml of the surrounding water. Using qPCR, we quantified higher values of faecal indicators which included inactive and dead microorganisms, detecting up to 2.6 × 102 gc mm-2. The ARGs were detected in 67-88% of the floating plastics and in 29-57% of the sediment plastics with a concentration of up to 6.7 × 102 gc mm-2. Furthermore, enrichment of these genes was observed in biofilms compared with the surrounding water. These results show that floating plastics act as a conduit for both the attachment and transport of faecal microorganisms. In contrast, low presence of faecal indicators was detected in plastic from seafloor sediments. Therefore, although in low concentrations, faecal bacteria, and potential pathogens, were identified in marine plastics, further suggesting plastics act as a reservoir of pathogens and ARGs.

A baseline assessment of the relationship between microplastics and plasticizers in sediment samples collected from the Barcelona continental shelf.

It has been suggested that the seafloor may be a sink for the plastic debris that enters the ocean. Therefore, the collection of data in the seafloor sediments regarding the co-presence of microplastics (MPs) and contaminants associated to plastic is considered a relevant topic. However, the number of studies addressing their possible correlation in this environment is still limited, and very little is known about the mechanisms that determine the release of plastic additives from plastic items. Starting from this basis, we investigated the presence of MPs and eleven phthalic acid esters (PAEs) in the continental shelf offshore Barcelona. Following a shelf-slope continuum approach, we sampled sediments from five stations, and we performed analysis by means of infrared micro spectroscopy (µFTIR) and liquid chromatography tandem mass spectrometry (LC-MS/MS). MPs were found to range from 62.0 to 931.1 items/kg d.w. with maximum concentration in the submarine canyon Besòs and at the highest depth. Moreover, different trends in the size distribution of fibers and non-fibers were observed, indicating the occurrence of a size dependent selection mechanism during transport and accumulation. PAEs resulted comprised between 1.35 to 2.41 mg/kg with Di(2-ethylhexyl)phthalate (DEHP) the most abundant congeners (1.04 mg/kg). Statistical analysis revealed no correlation between the Σ11PAEs and the total MPs concentration, but correlation between DEHP and fibers (σ = 0.667, p = 0,037), that resulted both correlated to the distance to the coast (ρ = 0.941 with p = 0,008 and ρ = 0.673 with p = 0.035, respectively).

Immunohistochemical Algorithm for the Classification of Muscle-Invasive Urinary Bladder Carcinoma with Lymph Node Metastasis: An Institutional Study.

Muscle-invasive urothelial carcinoma represents 20% of newly diagnosed cases of bladder cancer, and most cases show aggressive biological behavior with a poor prognosis. It is necessary to identify biomarkers that can be used as prognostic and predictive factors in daily clinical practice. In our study, we analyzed different antibodies in selected cases of muscle-invasive urinary bladder carcinoma and lymph node metastasis to identify immunohistochemical types and their value as possible prognostic factors. A total of 38 patients were included, 87% men and 13% women, with a mean age of 67.8 years. The most frequent histopathological type was urothelial carcinoma. In the primary lesion, the mixed type was the most common. In unilateral metastasis, the mixed type was the most frequently found. In cases of primary lesions and bilateral metastasis, the luminal and mixed types were observed. The luminal subtype was the most stable in immunohistochemical expression across primary tumors and metastases. The basal type showed a better prognosis in terms of disease-free survival. In conclusion, immunohistochemical studies are useful in assessing primary and metastatic lesions in patients with urothelial carcinoma. Immunohistochemical classification can typify muscle-invasive urothelial carcinoma, and the immunophenotype seems to have prognostic implications.

Decoding molecular programs in melanoma brain metastases.

Melanoma brain metastases (MBM) variably respond to therapeutic interventions; thus determining patient's prognosis. However, the mechanisms that govern therapy response are poorly understood. Here, we use a multi-OMICS approach and targeted sequencing (TargetSeq) to unravel the programs that potentially control the development of progressive intracranial disease. Molecularly, the expression of E-cadherin (Ecad) or NGFR, the BRAF mutation state and level of immune cell infiltration subdivides tumors into proliferative/pigmented and invasive/stem-like/therapy-resistant irrespective of the intracranial location. The analysis of MAPK inhibitor-naive and refractory MBM reveals switching from Ecad-associated into NGFR-associated programs during progression. NGFR-associated programs control cell migration and proliferation via downstream transcription factors such as SOX4. Moreover, global methylome profiling uncovers 46 differentially methylated regions that discriminate BRAFmut and wildtype MBM. In summary, we propose that the expression of Ecad and NGFR sub- classifies MBM and suggest that the Ecad-to-NGFR phenotype switch is a rate-limiting process which potentially indicates drug-response and intracranial progression states in melanoma patients.

A systematic review on microplastic pollution in water, sediments, and organisms from 50 coastal lagoons across the globe.

Coastal lagoons are transitional environments between continental and marine aquatic systems. Globally, coastal lagoons are of great ecological and socioeconomic importance as providers of valuable ecosystem services. However, these fragile environments are subject to several human pressures, including pollution by microplastics (MPs). The aim of this review was to identify and summarize advances in MP pollution research in coastal lagoons across the world. We consider peer-reviewed publications on this topic published in English and Spanish between 2000 and April 21, 2022, available in Scopus and Google Scholar. We found 57 publications with data on MP abundances and their characteristics in 50 coastal lagoons from around the world, 58% of which have some environmental protection status. The number of publications on this type of pollution in lagoons has increased significantly since 2019. Methodological differences amongst studies of MPs in coastal lagoons were nevertheless a limiting factor for wide-ranging comparisons. Most studies (77%) were conducted in single environmental compartments, and integration was limited, hampering current understanding of MP dynamics in such lagoons. MPs were more abundant in lagoons with highly populated shores and watersheds, which support intensive human activities. On the contrary, lagoons in natural protected areas had lower abundances of MPs, mostly in sediments and organisms. Fiber/filament and fragment shapes, and polyethylene, polyester, and polypropylene polymers were predominant. MPs had accumulated in certain areas of coastal lagoons, or had been exported to the sea, depending on the influence of seasonal weather, hydrodynamics, anthropogenic pressures, and typology of MPs. It is advised that future research on MP pollution in coastal lagoons should focus on methodological aspects, assessment/monitoring of pollution itself, MP dynamics and impacts, and prevention measures as part of a sound environmental management.

Impact of resuspended mine tailings on benthic biodiversity and ecosystem processes: The case study of Portmán Bay, Western Mediterranean Sea, Spain.

Industrial seabed mining is expected to cause significant impacts on marine ecosystems, including physical disturbance and the generation of plumes of toxin-laden water. Portmán Bay (NW Mediterranean Sea), where an estimated amount of 60 Mt of mine tailings from sulphide ores were dumped from 1957 to 1990, is one of the most metal-polluted marine areas in Europe and worldwide. This bay can be used to assess the impact on marine ecosystems of particle settling from sediment plumes resulting from mine tailings resuspension. With this purpose in mind, we conducted a field experiment there to investigate subsequent effects of deposition of (artificially resuspended) contaminated sediments on (i) prokaryotic abundance and meiofaunal assemblages (in terms of abundance and diversity), (ii) the availability of trophic resources (in terms of organic matter biochemical composition), and (iii) a set of ecosystem functions including meiofaunal biomass, heterotrophic C production and C degradation rates. The results of this study show that mine tailings resuspension and plume deposition led to the decline of prokaryotic abundance and nematode's biodiversity. The later decreased because of species removal and transfer along with particle resuspension and plume deposition. Such changes were also associated to a decrease of the proteins content in the sediment organic matter, faster C degradation rates and higher prokaryotic C production. Overall, this study highlights that mine tailing resuspension and ensuing particle deposition can have deleterious effects on both prokaryotes and nematode diversity, alter biogeochemical cycles and accelerate C degradation rates. These results should be considered for the assessment of the potential effects of seabed mineral exploitation on marine ecosystems at large.

Marine biofouling organisms on beached, buoyant and benthic plastic debris in the Catalan Sea.

Plastic debris provides long-lasting substrates for benthic organisms, thus acting as a potential vector for their dispersion. Its interaction with these colonizers is, however, still poorly known. This study examines fouling communities on beached, buoyant and benthic plastic debris in the Catalan Sea (NW Mediterranean), and characterizes the plastic type. We found 14 specimens belonging to two phyla (Annelida and Foraminifera) on microplastics, and more than 400 specimens belonging to 26 species in 10 phyla (Annelida, Arthropoda, Brachiopoda, Bryozoa, Chordata, Cnidaria, Echinodermata, Mollusca, Porifera and Sipuncula) on macroplastics. With 15 species, bryozoans are the most diverse group on plastics. We also report 17 egg cases of the catshark Scyliorhinus sp., and highlight the implications for their dispersal. Our results suggest that plastic polymers may be relevant for distinct fouling communities, likely due to their chemical structure and/or surface properties. Our study provides evidence that biofouling may play a role in the sinking of plastic debris, as the most abundant fouled plastics had lower densities than seawater, and all bryozoan species were characteristic of shallower depths than those sampled. More studies at low taxonomic level are needed in order to detect new species introduction and potential invasive species associated with plastic debris.

Tracking of Melanoma Cell Plasticity by Transcriptional Reporters.

Clonal evolution and cellular plasticity are the genetic and non-genetic driving forces of tumor heterogeneity, which in turn determine tumor cell responses towards therapeutic drugs. Several lines of evidence suggest that therapeutic interventions foster the selection of drug-resistant neural crest stem-like cells (NCSCs) that establish minimal residual disease (MRD) in melanoma. Here, we establish a dual-reporter system, enabling the tracking of NGFR expression and mRNA stability and providing insights into the maintenance of NCSC states. We observed that a transcriptional reporter that contained a 1-kilobase fragment of the human NGFR promoter was activated only in a minor subset (0.72 ± 0.49%, range 0.3-1.5), and ~2-4% of A375 melanoma cells revealed stable NGFR mRNA. The combination of both reporters provides insights into phenotype switching and reveals that both cellular subsets gave rise to cellular heterogeneity. Moreover, whole transcriptome profiling and gene-set enrichment analysis (GSEA) of the minor cellular subset revealed hypoxia-associated programs that might serve as potential drivers of an in vitro switching of NGFR-associated phenotypes and relapse of post-BRAF inhibitor-treated tumors. Concordantly, we observed that the minor cellular subset increased in response to dabrafenib over time. In summary, our reporter-based approach provides insights into plasticity and identified a cellular subset that might be responsible for the establishment of MRD in melanoma.