Nova Medical (United States)

Dear Editor, Over the past two decades, the role of the cardiorespiratory physiotherapist has become increasingly prominent within clinical care pathways, significantly contributing to the management of both acute and chronic respiratory diseases...

Background Cutaneous melanoma (CM) is the most aggressive form of skin cancer, with an increasing incidence in recent years.Immunotherapy is considered the most promising therapeutic option for late-stage CM.However, resistance occurs in 40-65% of the cases and 20-30% of responding patients develop secondary resistance, mainly due to the immunosuppression imposed by the tumor microenvironment.For this reason, it is essential to investigate the complex interactions between immune cells and tumors.Methods So far, most of the studies in this context have relied on in vivo models.However, these models present both scientific and ethical limitations.In our lab, we have established an in vitro CM skin model that fully recapitulates skin pathophysiology.Therefore, our objective is now to optimize an immune competent in vitro CM skin model to study the crosstalk between tumor cells and the immune system, and to investigate the mechanisms underlying immunotherapy response and resistance.For this, we developed an in vitro CM skin model composed by a dermis equivalent made with fibroblasts and the extracellular matrix they secrete, and an epidermis equivalent composed by CM cells and keratinocytes that stratify into all the layers of the epidermis.Once the model was fully established, we added to the medium PBMCs previously isolated from healthy donors.To determine the best conditions that allow the immune cells to infiltrate the skin equivalent and migrate toward the CM cells, we tested different immune/CM cell ratios (3:1; 10:1; 30:1) and different incubation times (2, 3 and 4 days).In parallel, as a control condition, we performed the same model but without immune cells.After fixation, the models have been processed for paraffin embedding (for histological analysis with haematoxylin and eosin staining (H&E)) and cryopreservation (for immunofluorescence microscopy analysis).Results It is possible to recognize by H&E staining the presence of small round cells in the dermis of the model to which PBMCs were added for 4 days at a 30:1 ratio of PBMCs/CM cells, consistent with the morphology of this type of cells.This observation suggests that there was infiltration of PBMCs into the CM skin model.However, since H&E staining is not sufficient to discriminate between PBMCs and the other cell types, we intend to perform immunofluorescence using an antibody that recognizes CD45, a marker that is specifically expressed by PBMCs. ConclusionThe results obtained are promising due to the presence in the dermis of small, rounded cells that are morphologically compatible with PBMCs, suggesting their infiltration in the model.However, immunofluorescence staining is essential to confirm the results.Once established, this model will provide a clinically relevant platform to identify predictive biomarkers of immunotherapy response and novel therapeutic strategies to overcome immunotherapy resistance in CM.P08.

Kidney transplant rejection presents a clinical challenge, requiring an accurate and timely diagnosis. While kidney graft biopsy remains the gold standard, its inherent limitations motivate the need for noninvasive diagnosis. This review comprehensively examines emerging non-invasive biomarkers for the monitoring of kidney allograft, specifically donor-derived cell-free DNA (dd-cfDNA), gene expression profiling (GEP), microRNAs (miRNAs) and long non-coding RNAs (lncRNAs), chemokines, and DNA methylation patterns. We review their underlying biological principles and discuss their diagnostic accuracy, clinical applications, and limitations in the current available evidence. Donor-derived cfDNA, a promising predictor of allograft injury, shows particular utility in detecting antibody-mediated rejection (ABMR), lacking however for standardization concerning its cutoff. Gene expression profiling provides insights into immune responses, with various gene signatures developed for detecting subclinical and acute rejection. MicroRNAs and long non-coding RNAs demonstrate potential as tissue-specific biomarkers of injury and rejection, yet face challenges in standardization and clinical translation. Chemokines like urinary CXCL9 and CXCL10 exhibit diagnostic and prognostic value for acute rejection. Furthermore, DNA methylation patterns offer a novel approach to identify cell-specific cfDNA and reflect cell death processes within transplanted organs. The integration of these diverse biomarkers, potentially augmented by artificial intelligence platforms, holds immense promise for improving diagnostic precision, minimizing the need for biopsies, and enabling personalized immunosuppression strategies, which would improve long-term graft survival.

Long COVID (LC) is a post-infectious condition affecting millions worldwide, characterized by persistent multisystem symptoms. Females are disproportionately affected, reporting higher symptom burden, particularly neurocognitive and neurosensory complaints. While short-term immunopathology has been described, the long-term clinical course, immune dysregulation, and sex-specific underpinnings remain poorly understood. We analyzed 34 participants experiencing persisting symptoms from 9months to 5years post-SARS-CoV-2 infection, alongside 26 SARS-CoV-2-infected controls without symptoms. Clinical assessments, symptom inventories, comorbidity analysis, and work capacity evaluation were performed. Immune profiling included flow cytometry of CD4⁺ and CD8⁺ T cells, NK cells, and B cells, as well as quantification of plasma cytokines, soluble factors, and cytotoxic molecules, analyzed in a sex-disaggregated manner. Females with LC exhibited higher symptom burden, particularly persistent fatigue, neurocognitive and neurosensory complaints, which increased with age and tended to increase with disease duration, whereas males showed no clear age- or duration-related patterns. Comorbidities, especially affecting endocrine, metabolic, and circulatory systems, were more frequent in females and aligned with symptom severity. Immune profiling revealed subtle but sex-specific differences: females had reduced CD8⁺ T cell cytotoxic profile, lower NKG2D and granzyme K expression, increased sCD40L and sFAS, and decreased perforin, whereas males displayed elevated TNF-α. NK cell function, B cells, and humoral immunity remained largely intact. Over half of participants reported functional impairments affecting work capacity. Even though our cohort is small it suggests that prolonged LC is characterized by sex-specific differences in symptom burden and immune profiles. Reduced cytotoxic CD8⁺ T cell profile in females may contribute to viral persistence and neurological symptoms, whereas elevated inflammatory markers in males suggest distinct immune pathways. These findings highlight the need for sex- and duration-specific management strategies, the identification of biomarkers, and the development of personalized therapies targeting specific LC endotypes.

This article presents a systematic review on methods for quantifying three-dimensional, time-resolved (3D+t) deformation and motion of human arteries from Computed Tomography (CT) and Magnetic Resonance Imaging (MRI). Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, we searched Scopus, Web of Science, IEEE Xplore, Google Scholar, and PubMed on 19 December 2025 for in vivo, patient-specific CT or MRI studies reporting motion or deformation of large human arteries. We included studies that quantified arterial deformation or motion tracking and excluded non-vascular tissues, in vitro or purely computational work. Thirty-five studies were included in the qualitative synthesis; most were small, single-centre observational cohorts. Articles were analysed qualitatively, and results were synthesised narratively. Across the 35 studies, the most common segmentation approaches are active contours and threshold, while temporal motion is tracked using either voxel registration or surface methods. These kinematic data are used to compute metrics such as circumferential and longitudinal strain, distensibility, and curvature. Several studies also employ inverse methods to estimate wall stiffness. The findings consistently show that arterial strain decreases with age (on the order of 20% per decade in some cases) and in the presence of disease, that stiffness correlates with geometric remodelling, and that deformation is spatially heterogeneous. However, insufficient data prevents meaningful comparison across methods.

El fenotipo sésil en Candida spp., confiere una resistencia mayor que la de sus contrapartes planctónicas, en el que los resultados de la Concentración Mínima Inhibitoria (CMI) obtenidos por los métodos estándar no se correlacionen con la eficacia clínica. Se validó el uso de la resazurina como un indicador colorimétrico eficiente, rápido y económico para determinar la verdadera resistencia de las biopelículas, mediante CMI en su estado sésil, en Candida spp., aisladas de hemocultivos. La metodología implicó la determinación de la CMI planctónica mediante microdilución en caldo con lectura visual y la determinación de la CMI en su estado sésil, utilizando resazurina como indicador colorimétrico con lectura visual en especies de Candida, donde la solución de resazurina actúa como un indicador metabólico que reduce el color azul a rosa exponiendo la viabilidad celular dentro de la biopelícula tras la exposición al fármaco, en este caso fluconazol. Todos los aislados tenían una capacidad de débil a moderada para formar biopelículas, la comparación entre los dos estados fenotípicos demostró una disparidad crítica en la susceptibilidad al fluconazol, mientras que la CMI de las células planctónica se mantuvo ≤ 0.5 μg/mL, la CMI sésil refirió CMIs desde 512 μg/mL hasta 8.500 μg/mL a la CMI inicial de las mismas. Se concluye que el método basado en la utilización de la resazurina es un sistema robusto y asequible para la determinación de la CMI en su estado sésil, siendo una herramienta diagnóstica fundamental para guiar de manera más precisa la terapia antifúngica y mejorar el pronóstico de los pacientes.

Abstract Background and Aims Sjögren's Syndrome (SSj) is an autoimmune disease that primarily affects exocrine glands. The renal manifestations of SSj are diverse, with tubulointerstitial nephritis being the most common form of kidney damage. Although renal involvement is rare, it can lead to chronic kidney disease (CKD). Understanding the factors that predict renal impairment in SSj is crucial for early intervention and improving patient outcomes. This study aims to investigate the prevalence of CKD stages in SSj and identify the factors associated with worse renal outcomes in a retrospective cohort. Method This retrospective study reviewed patients diagnosed with SSj at an autoimmune outpatient clinic in a Portuguese center from 2013 to 2024. Data on clinical symptoms, immunological markers, and kidney function parameters were collected. The prevalence of CKD stages was assessed using estimated glomerular filtration rate (eGFR) values. Univariate statistical analyses were performed to assess the association between clinical and laboratory variables, including the presence of autoantibodies and comorbidities and CKD progression. Multivariate analyses were performed, including variables that theoretically impact the outcome of interest and variables which had statistical significance in the univariate analyses. Results Out of 158 identified SSj patients, 133 were included after excluding those lost to follow-up or deceased. The cohort predominantly comprised women (96.2%) with a mean age of 67 ± 14 years and an average disease duration of 11 ± 7 years. The most commonly reported symptoms were xerophthalmia (97%), xerostomia (82%) and arthritis (46%). The Chisholm-Mason biopsy scores showed that 36 patients scored below 2, 15 scored 3 and 21 scored 4; 61 patients had no biopsy records. Positive autoantibodies were frequently found, with ANA positivity in 114 patients(86%), anti-SSA positivity in 80 patients(60%), and anti-SSB positivity in 39 patients(29%). Hypergammaglobulinemia was present in 29 patients, and hypocomplementemia was identified in 14 patients. Comorbidities were present in 48.8% patients, including hypertension (43.6%), dyslipidemia (48.1%) and type 2 diabetes mellitus (10.5%). Systemic immunosuppression was used in 81 patients (60%). Four patients were under nephrology follow-up: three cases were related to SSj, including one with tubulointerstitial nephritis and two with end-stage CKD of unknown etiology. No other types of renal involvement were documented. Pulmonary involvement was found in three patients. Among the cohort, 86 patients (64.6%) had no other associated autoimmune diseases. The mean eGFR was 85 ml/min/1.73 m². The CKD staging is presented in Table 1. Proteinuria above 30 mg/g was observed in 31 patients, with only one patient presenting a urine protein/creatinine ratio greater than 500 mg/g. We performed univariate correlation analysis for eGFR. We found associations with hypocomplementemia (r:0.2097; P = 0.0158), the presence of SSA (r:0.1775; P = 0.0418) and with the presence of hypertension (r:0.2826; P = 0.0010). We found no associations with clinical manifestations of SSj, the presence of other antibodies, or with any treatment. In multivariate analysis, in a model which included the presence of hypertension, diabetes, xerostomia, xerophthalmia, SSA, SSB, ANA, hypocomplementemia, proteinuria, and disease duration, only hypocomplementemia remained statistically significant (B = 12.25 [0.67–23.84]; P = 0.038). When analyzing CKD stages, proteinuria and proteinuria stages, univariate correlations did not persist when evaluating in multivariate models. Conclusion Chronic kidney disease is a rare but significant manifestation of sjögren's syndrome. Early stages of CKD are often asymptomatic, making it crucial to identify at-risk patients before irreversible damage occurs. Hypocomplementemia is a risk factor for decreased eGFR, suggesting that ongoing inflammation has a detrimental effect on kidney function.