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A 13-year-old male presented with a 2-year history of recurrent aphthous stomatitis. The patient had undergone several unsuccessful treatments, leading to the decision to initiate apremilast. He showed a good clinical response with reduction in outbreaks, and no adverse effects were observed. This is the first report, to our knowledge, of a pediatric patient treated with apremilast for this indication.
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Introduction Patient-derived organoids have emerged as a promising in vitro model for precision medicine, particularly in cancer, but also in non-cancer-related diseases. However, the optimal culture medium for culturing patient-derived lung organoids has not yet been agreed upon. This study aimed to shed light on the optimal selection of a culture media for developing studies using patient-derived lung organoids. Methods Tumor and normal paired tissue from 71 resected non-small cell lung cancer patients were processed for organoid culture. Lung cancer organoids (LCOs) were derived from tumor tissue and normal lung organoids (LNOs) from non-neoplastic lung tissue. Three different culture media were compared: Permissive culture medium (PCM); Limited culture medium (LCM); and Minimum basal medium (MBM). We assessed their effectiveness in establishing organoid cultures, promoting organoid growth and viability, and compared their differential phenotypic characteristics. Results While PCM was associated with the highest success rate and useful for long-term expansion, MBM was the best option to avoid normal organoid overgrowth in the organoid culture. The density, size, and viability of LNOs organoids were reduced using LCM and severely affected with MBM. LNOs cultured in PCM tend to differentiate to bronchospheres, while alveolosphere differentiation can be observed in those cultured with LCM. The morphological phenotype of LCO was influenced by the culture media of election. Mesenchymal cell overgrowth was observed when LCM was used. Conclusion This work highlights the importance of considering the research objectives when selecting the most suitable culture medium for growing patient-derived lung organoids.
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This study aimed to determine the relationship between geometric and dosimetric agreement metrics in head and neck (H&N) cancer radiotherapy plans. A total 287 plans were retrospectively analyzed, comparing auto-contoured and clinically used contours using a Dice similarity coefficient (DSC), surface DSC (sDSC), and Hausdorff distance (HD). Organs-at-risk (OARs) with ≥200 cGy dose differences from the clinical contour in terms of Dmax (D0.01cc) and Dmean were further examined against proximity to the planning target volume (PTV). A secondary set of 91 plans from multiple institutions validated these findings. For 4995 contour pairs across 19 OARs, 90% had a DSC, sDSC, and HD of at least 0.75, 0.86, and less than 7.65 mm, respectively. Dosimetrically, the absolute difference between the two contour sets was <200 cGy for 95% of OARs in terms of Dmax and 96% in terms of Dmean. In total, 97% of OARs exhibiting significant dose differences between the clinically edited contour and auto-contour were within 2.5 cm PTV regardless of geometric agreement. There was an approximately linear trend between geometric agreement and identifying at least 200 cGy dose differences, with higher geometric agreement corresponding to a lower fraction of cases being identified. Analysis of the secondary dataset validated these findings. Geometric indices are approximate indicators of contour quality and identify contours exhibiting significant dosimetric discordance. For a small subset of OARs within 2.5 cm of the PTV, geometric agreement metrics can be misleading in terms of contour quality.
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Interleukin (IL)-9 is present in atopic dermatitis (AD) lesions and is considered to be mainly produced by skin-homing T cells expressing the cutaneous lymphocyte-associated antigen (CLA). However, its induction by AD-associated triggers remains unexplored. Circulating skin-tropic CLA+ and extracutaneous/systemic CLA- memory T cells cocultured with autologous lesional epidermal cells from AD patients were activated with house dust mite (HDM) and staphylococcal enterotoxin B (SEB). Levels of AD-related mediators in response to both stimuli were measured in supernatants, and the cytokine response was associated with different clinical characteristics. Both HDM and SEB triggered heterogeneous IL-9 production by CLA+ and CLA- T cells in a clinically homogenous group of AD patients, which enabled patient stratification into IL-9 producers and non-producers, with the former group exhibiting heightened HDM-specific and total IgE levels. Upon allergen exposure, IL-9 production depended on the contribution of epidermal cells and class II-mediated presentation; it was the greatest cytokine produced and correlated with HDM-specific IgE levels, whereas SEB mildly induced its release. This study demonstrates that both skin-tropic and extracutaneous memory T cells produce IL-9 and suggests that the degree of allergen sensitization reflects the varied IL-9 responses in vitro, which may allow for patient stratification in a clinically homogenous population.
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Dermatitis Atópica , Enterotoxinas , Interleucina-9 , Células T de Memoria , Dermatitis Atópica/inmunología , Dermatitis Atópica/metabolismo , Humanos , Interleucina-9/metabolismo , Femenino , Masculino , Adulto , Enterotoxinas/inmunología , Células T de Memoria/inmunología , Células T de Memoria/metabolismo , Piel/inmunología , Piel/metabolismo , Pyroglyphidae/inmunología , Animales , Inmunoglobulina E/inmunología , Inmunoglobulina E/sangre , Persona de Mediana Edad , Antígenos de Diferenciación de Linfocitos T/metabolismo , Adulto Joven , Alérgenos/inmunología , Adolescente , Glicoproteínas de MembranaRESUMEN
B-cell epidermotropism is an exceptional finding in cutaneous lymphomas. The few cases previously described in the literature mostly correspond to systemic lymphomas, with the most frequent being splenic marginal zone lymphoma. This lymphoma can manifest as a cutaneous eruption preceding the splenomegaly, with systemic involvement demonstrated by bone marrow and/or peripheral blood. This presentation is known as epidermotropic B-cell lymphoma. Herein, we present a new case that emphasizes B-cell epidermotropism as a feature that should alert the clinician of the possibility of a secondary involvement and, consequently, prompt them to expand the recommended initial staging.
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BACKGROUND: During pseudoglandular stage of the human lung development the primitive bronchial buds are initially conformed by simple tubules lined by endoderm-derived epithelium surrounded by mesenchyme, which will progressively branch into airways and start to form distal epithelial saculles. For first time alveolar type II (AT2) pneumocytes appears. This study aims to characterize the genes and microRNAs involved in this differentiation process and decipher its role in the starting alveolar differentiation. METHODS: Gene and microRNA profiling was performed in human embryonic lungs from 7 to 12 post conception weeks (pcw). Protein expression location of candidate genes were analyzed by immunofluorescense in embryonic lung tissue sections. mRNA/miRNA target pairs were identified using computational approaches and their expression was studied in purified epithelial/mesenchymal cell populations and in isolated tips and stalks from the bronchial tree. Additionally, silencing experiments in human embryonic lung mesenchymal cells and in human embryonic tip-derived lung organoids were performed, as well as organoid differentiation studies. AT2 cell markers were studied by qRT-PCR and by immunofluorescence. The TGFB-ß phosphorylated pathways was analyzed with membrane protein arrays. Lung explants were cultured in air/liquid interface with/without peptides. RESULTS: We identified 88 differentially expressed genes, including IGFBP3. Although IGFBP3 mRNA was detected in both epithelial and mesenchymal populations, the protein was restricted to the epithelium, indicating post-transcriptional regulation preventing IGFBP3 protein expression in the mesenchyme. MicroRNA profiling identified miR-34a as an IGFBP3 regulator. miR-34a was up-regulated in mesenchymal cells, and its silencing in human embryonic lung mesenchymal cells increased IGFBP3 levels. Additionally, IGFBP3 expression showed a marked downregulation from 7 to 12 pcw, suggesting its involvement in the differentiation process. The differentiation of human tip-derived lung embryonic organoids showed a drastic reduction in IGFBP3, supported by the scRNAseq data. IGFBP3 silencing in organoids activated an alveolar-like differentiation process characterized by stem cell markers downregulation and upregulation of AT2 markers. This process was mediated by TGFß signalling inhibition and BMP pathway activation. CONCLUSIONS: The IGFBP3/miR-34a axis restricts IGFBP3 expression in the embryonic undifferentiated lung epithelium, and the progressive downregulation of IGFBP3 during the pseudoglandular stage is required for alveolar differentiation.
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Diferenciación Celular , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina , Pulmón , MicroARNs , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/genética , Pulmón/metabolismo , Pulmón/embriología , Pulmón/citología , Células Epiteliales Alveolares/metabolismo , Células Epiteliales Alveolares/citología , Alveolos Pulmonares/metabolismo , Alveolos Pulmonares/citología , Regulación del Desarrollo de la Expresión Génica , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/citologíaRESUMEN
A healthy young male patient was referred to the department of dermatology for evaluation of a solitary painful scrotal ulceration that developed rapidly 48 hours before consultation. What is your diagnosis?
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Necrosis , Escroto , Humanos , Masculino , Escroto/patología , Necrosis/diagnóstico , Enfermedad Aguda , Úlcera Cutánea/patología , Úlcera Cutánea/diagnóstico , Úlcera Cutánea/microbiología , Adulto , BiopsiaRESUMEN
Inflammatory Bowel Disease (IBD) comprises a heterogeneous group of chronic inflammatory conditions of the gastrointestinal tract that include ulcerative colitis (UC) and Crohn's disease. Although the etiology is not well understood, IBD is characterized by a loss of the normal epithelium homeostasis that disrupts the intestinal barrier of these patients. Previous work by our group demonstrated that epithelial homeostasis along the colonic crypts involves a tight regulation of lipid profiles. To evaluate whether lipidomic profiles conveyed the functional alterations observed in the colonic epithelium of IBD, we performed matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) analyses of endoscopic biopsies from inflamed and non-inflamed segments obtained from UC patients. Our results indicated that lipid profiling of epithelial cells discriminated between healthy and UC patients. We also demonstrated that epithelial cells of the inflamed mucosa were characterized by a decrease in mono- and di-unsaturated fatty acid-containing phospholipids and higher levels of arachidonic acid-containing species, suggesting an alteration of the lipid gradients occurring concomitantly to the epithelial differentiation. This result was reinforced by the immunofluorescence analysis of EPHB2 and HPGD, markers of epithelial cell differentiation, sustaining that altered lipid profiles were at least partially due to a faulty differentiation process. Overall, our results showed that lipid profiling by MALDI-MSI faithfully conveys molecular and functional alterations associated with the inflamed epithelium, providing the foundation for a novel molecular characterization of UC patients.
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Diferenciación Celular , Colon , Humanos , Colon/metabolismo , Colon/patología , Masculino , Femenino , Colitis Ulcerosa/metabolismo , Colitis Ulcerosa/patología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Persona de Mediana Edad , Adulto , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Lipidómica/métodos , Enterocitos/metabolismo , Enterocitos/patología , Metabolismo de los Lípidos , Inflamación/metabolismo , Inflamación/patología , Lípidos/análisis , Células Epiteliales/metabolismo , Células Epiteliales/patologíaRESUMEN
Schistosomiasis is a neglected disease of poverty that affects over 200 million people worldwide and relies on a single drug for therapy. The cathepsin B1 cysteine protease (SmCB1) of Schistosoma mansoni has been investigated as a potential target. Here, a structure-based pharmacophore virtual screening (VS) approach was used on a data set of approved drugs to identify potential antischistosomal agents targeting SmCB1. Pharmacophore (PHP) models underwent validation through receiver operating characteristics curves achieving values >0.8. The data highlighted riboflavin (RBF) as a compound of particular interest. A 1 µs molecular dynamics simulation demonstrated that RBF altered the conformation of SmCB1, causing the protease's binding site to close around RBF while maintaining the protease's overall integrity. RBF inhibited the activity of SmCB1 at low micromolar values and killed the parasite in vitro. Finally, in a murine model of S. mansoni infection, oral administration of 100 mg/kg RBF for 7 days significantly decreased worm burdens by â¼20% and had a major impact on intestinal and fecal egg burdens, which were decreased by â¼80%.
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Anticuerpos Monoclonales Humanizados , Humanos , Anticuerpos Monoclonales Humanizados/efectos adversos , Anticuerpos Monoclonales Humanizados/uso terapéutico , Masculino , Femenino , Anciano , Erupciones por Medicamentos/etiología , Erupciones por Medicamentos/diagnóstico , Persona de Mediana EdadRESUMEN
BACKGROUND: Surgical risk stratification is crucial for enhancing perioperative assistance and allocating resources efficiently. However, existing models may not capture the complexity of surgical care in Brazil. Using data from various healthcare settings nationwide, we developed a new risk model for 30-day in-hospital mortality (the Ex-Care BR model). METHODS: A retrospective cohort study was conducted in 10 hospitals from different geographic regions in Brazil. Data were analysed using multilevel logistic regression models. Model performance was assessed using the area under the receiver operating characteristic curve (AUROC), Brier score, and calibration plots. Derivation and validation cohorts were randomly assigned. RESULTS: A total of 107,372 patients were included, and 30-day in-hospital mortality was 2.1% (n=2261). The final risk model comprised four predictors related to the patient and surgery (age, ASA physical status classification, surgical urgency, and surgical size), and the random effect related to hospitals. The model showed excellent discrimination (AUROC=0.93, 95% confidence interval [CI], 0.93-0.94), calibration, and overall performance (Brier score=0.017) in the derivation cohort (n=75,094). Similar results were observed in the validation cohort (n=32,278) (AUROC=0.93, 95% CI, 0.92-0.93). CONCLUSIONS: The Ex-Care BR is the first model to consider regional and organisational peculiarities of the Brazilian surgical scene, in addition to patient and surgical factors. It is particularly useful for identifying high-risk surgical patients in situations demanding efficient allocation of limited resources. However, a thorough exploration of mortality variations among hospitals is essential for a comprehensive understanding of risk. CLINICAL TRIAL REGISTRATION: NCT05796024.
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Mortalidad Hospitalaria , Humanos , Masculino , Femenino , Brasil/epidemiología , Persona de Mediana Edad , Estudios Retrospectivos , Anciano , Medición de Riesgo/métodos , Adulto , Procedimientos Quirúrgicos Operativos/mortalidad , Estudios de Cohortes , Anciano de 80 o más Años , Curva ROC , Adulto Joven , Factores de RiesgoAsunto(s)
Urticaria Crónica , Síndromes Periódicos Asociados a Criopirina , Errores Diagnósticos , Mosaicismo , Proteína con Dominio Pirina 3 de la Familia NLR , Humanos , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Síndromes Periódicos Asociados a Criopirina/diagnóstico , Síndromes Periódicos Asociados a Criopirina/genética , Urticaria Crónica/diagnóstico , Urticaria Crónica/genética , Femenino , Adulto , Masculino , MutaciónRESUMEN
In this proof-of-concept study, spatial transcriptomics combined with public single-cell ribonucleic acid-sequencing data were used to explore the potential of this technology to study kidney allograft rejection. We aimed to map gene expression patterns within diverse pathologic states by examining biopsies classified across nonrejection, T cell-mediated acute rejection, interstitial fibrosis, and tubular atrophy. Our results revealed distinct immune cell signatures, including those of T and B lymphocytes, monocytes, mast cells, and plasma cells, and their spatial organization within the renal interstitium. We also mapped chemokine receptors and ligands to study immune cell migration and recruitment. Finally, our analysis demonstrated differential spatial enrichment of transcription signatures associated with kidney allograft rejection across various biopsy regions. Interstitium regions displayed higher enrichment scores for rejection-associated gene expression patterns than tubular areas, which had negative scores. This implies that these signatures are primarily driven by processes unfolding in the renal interstitium. Overall, this study highlights the value of spatial transcriptomics for revealing cellular heterogeneity and immune signatures in renal transplant biopsies and demonstrates its potential for studying the molecular and cellular mechanisms associated with rejection. However, certain limitations must be borne in mind regarding the development and future applications of this technology.
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Rechazo de Injerto , Trasplante de Riñón , Prueba de Estudio Conceptual , Transcriptoma , Rechazo de Injerto/patología , Rechazo de Injerto/genética , Rechazo de Injerto/etiología , Trasplante de Riñón/efectos adversos , Humanos , Perfilación de la Expresión Génica , Pronóstico , Supervivencia de Injerto/inmunología , Biomarcadores/metabolismo , AloinjertosRESUMEN
Renal ischemia-reperfusion injury (IRI) leads to endoplasmic reticulum (ER) stress, thereby initiating the unfolded protein response (UPR). When sustained, this response may trigger the inflammation and tubular cell death that acts to aggravate the damage. Here, we show that knockdown of the BET epigenetic reader BRD4 reduces the expression of ATF4 and XBP1 transcription factors under ER stress activation. BRD4 is recruited to the promoter of these highly acetylated genes, initiating gene transcription. Administration of the BET protein inhibitor, JQ1, one hour after renal damage induced by bilateral IRI, reveals reduced expression of ATF4 and XBP1 genes, low KIM-1 and NGAL levels and recovery of the serum creatinine and blood urea nitrogen levels. To determine the molecular pathways regulated by ATF4 and XBP1, we performed stable knockout of both transcription factors using CRISPR-Cas9 and RNA sequencing. The pathways triggered under ER stress were mainly XBP1-dependent, associated with an adaptive UPR, and partially regulated by JQ1. Meanwhile, treatment with JQ1 downmodulated most of the pathways regulated by ATF4 and related to the pathological processes during exacerbated UPR activation. Thus, BRD4 inhibition could be useful for curbing the maladaptive UPR activation mechanisms, thereby ameliorating the progression of renal disease.
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Antineoplásicos , Daño por Reperfusión , Humanos , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteínas Nucleares/genética , Estrés del Retículo Endoplásmico/genética , Respuesta de Proteína Desplegada , Antineoplásicos/farmacología , Daño por Reperfusión/tratamiento farmacológico , Daño por Reperfusión/genética , Daño por Reperfusión/metabolismo , Proteínas que Contienen Bromodominio , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismoRESUMEN
This research examines the influence of adding a commercial ionic liquid to the electrolyte during the electrochemical anodization of tungsten for the fabrication of WO3 nanostructures for photoelectrochemical applications. An aqueous electrolyte composed of 1.5 M methanesulfonic acid and 5% v/v [BMIM][BF4] or [EMIM][BF4] was used. A nanostructure synthesized in an ionic-liquid-free electrolyte was taken as a reference. Morphological and structural studies of the nanostructures were performed via field emission scanning electron microscopy and X-ray diffraction analyses. Electrochemical characterization was carried out using electrochemical impedance spectroscopy and a Mott-Schottky analysis. From the results, it is highlighted that, by adding either of the two ionic liquids to the electrolyte, well-defined WO3 nanoplates with improved morphological, structural, and electrochemical properties are obtained compared to samples synthesized without ionic liquid. In order to evaluate their photoelectrocatalytic performance, the samples were used as photocatalysts to generate hydrogen by splitting water molecules and in the photoelectrochemical degradation of methyl red dye. In both applications, the nanostructures synthesized with the addition of either of the ionic liquids showed a better performance. These findings confirm the suitability of ionic liquids, such as [BMIM][BF4] and [EMIM][BF4], for the synthesis of highly efficient photoelectrocatalysts via electrochemical anodization.
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Purpose: Chemoradiation therapy (CRT) is the standard treatment for squamous cell carcinoma of the anus (SCCA). This study aimed to investigate the relationship between vaginal dosimetry and long-term patient-reported dyspareunia after treatment. We further aimed to use the anterior vaginal wall (AVW) as an organ at risk to define an actionable dosimetric clinical goal to decrease the risk of patient-reported dyspareunia. Methods and Materials: Women with SCCA treated with intensity modulated radiation therapy-based CRT were surveyed at least 2 years after successfully completing therapy. A Female Sexual Function Index (FSFI) pain subscore ≤4 was used to define dyspareunia. Dosimetric parameters were calculated for both the full vaginal canal and AVW. Multivariable linear regression models were created to identify predictors of FSFI pain subscore using backward selection to identify final variables include in the models. An actionable dosimetric predictor for dyspareunia was established using the Youden index method for cutoff optimization. Results: Of 184 women who were contacted, 90 (49%) returned completed surveys. Of those who completed surveys, 51 (56.7%) reported being sexually active, and 47 had dosimetric data available for review. Of sexually active respondents, 32 (68%) had an FSFI pain subscore ≤4. Multiple regression models were generated using the full vaginal canal and AVW as organs at risk, and both models showed similar predictive relationships with volumetric dose parameters emerging as the best dosimetric predictors for dysparenuia. Age over 65 years was also associated with higher FSFI pain subscores (eg, less pain with intercourse) in both models. AVW V35 Gy < 60% was identified as the optimal cutoff to reduce the risk of patient-reported dyspareunia. Conclusions: Increased dose to the vaginal canal is significantly associated with worse patient-reported dyspareunia following CRT for SCCA. Minimizing dose to the AVW to V35 Gy < 60% may reduce the risk of this quality of life-limiting toxicity. Further prospective evaluation is needed to validate these findings.
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BACKGROUND: Topical imiquimod has shown to be an effective treatment for EMPD, although available evidence supporting its use is based on case reports and small series of patients. OBJECTIVES: To investigate the therapeutic outcomes and analyze potential clinico-pathological factors associated with imiquimod response in a large cohort of EMPD patients. METHODS: Retrospective chart review of 125 EMPD patients treated with imiquimod at 20 Spanish tertiary-care hospitals. RESULTS: During the study period, patients received 134 treatment regimens with imiquimod, with 70 (52.2%) cases achieving complete response (CR), 41 (30.6%) partial response and 23 (17.2%) no response. The cumulative CR rates at 24 and 48 weeks of treatment were 46.3% and 71.8%, respectively, without significant differences between first-time and previously treated EMPD. Larger lesions (≥6â cm; p = 0.038) and EMPD affecting >1 anatomical site (p = 0.002) were significantly associated with a worse treatment response. However, the CR rate did not differ significantly by the number of treatment applications (≤4 vs. > 4 times/week; p = 0.112). Among patients who achieved CR, 30 (42.9%) developed local recurrences during a mean follow-up period of 36â months, with an estimated 3 and 5-year recurrence free-survival of 55.7% and 36.4%, respectively. CONCLUSIONS: Imiquimod appears as an effective therapeutic alternative for both first-line and previously treated EMPD lesions. However, a less favorable therapeutic response could be expected in larger lesions and those affecting >1 anatomical site. Based on our results, a 3-4 times weekly regimen of imiquimod with a treatment duration of at least 6â months could be considered an appropriate therapeutic strategy for EMPD patients.
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Targeting epigenetic mechanisms has emerged as a potential therapeutic approach for the treatment of kidney diseases. Specifically, inhibiting the bromodomain and extra-terminal (BET) domain proteins using the small molecule inhibitor JQ1 has shown promise in preclinical models of acute kidney injury (AKI) and chronic kidney disease (CKD). However, its clinical translation faces challenges due to issues with poor pharmacokinetics and side effects. Here, we developed engineered liposomes loaded with JQ1 with the aim of enhancing kidney drug delivery and reducing the required minimum effective dose by leveraging cargo protection. These liposomes efficiently encapsulated JQ1 in both the membrane and core, demonstrating superior therapeutic efficacy compared to freely delivered JQ1 in a mouse model of kidney ischemia-reperfusion injury. JQ1-loaded liposomes (JQ1-NPs) effectively targeted the kidneys and only one administration, one-hour after injury, was enough to decrease the immune cell (neutrophils and monocytes) infiltration to the kidney-an early and pivotal step to prevent damage progression. By inhibiting BRD4, JQ1-NPs suppress the transcription of pro-inflammatory genes, such as cytokines (il-6) and chemokines (ccl2, ccl5). This success not only improved early the kidney function, as evidenced by decreased serum levels of BUN and creatinine in JQ1-NPs-treated mice, along with reduced tissue expression of the damage marker, NGAL, but also halted the production of extracellular matrix proteins (Fsp-1, Fn-1, α-SMA and Col1a1) and the fibrosis development. In summary, this work presents a promising nanotherapeutic strategy for AKI treatment and its progression and provides new insights into renal drug delivery.