Histological healing is increasingly recognized as a sensitive marker of disease remission in ulcerative colitis (UC). However, the dynamics of mucosal T lymphocytes and proinflammatory cytokines during healing remain incompletely understood. In this paired, within-subject observational study (retrospective analysis of paired biopsies), colonic biopsy sets from 20 adult UC patients were analyzed during active inflammation and at a subsequent time point of histologic healing. Immunohistochemistry was performed for CD3, CD4, CD8, and IL-6. Lymphocyte densities were quantified in intraepithelial and lamina propria compartments, while IL-6 expression was scored semi-quantitatively. Histological activity was assessed using the Geboes score. Intraepithelial CD4+ T cells significantly decreased during histologic healing (mean 6.8 → 3.75 cells/100 epithelial cells, p < 0.05), whereas lamina propria CD4+ cells remained variably persistent, suggesting ongoing immune regulation. Intraepithelial CD8+ cells increased during remission, indicating a potential reparative or surveillance role. IL-6 expression markedly declined in epithelial and stromal compartments during healing, reflecting resolution of mucosal inflammation. Correlation analyses revealed enhanced coordination between CD4+ and CD8+ cells in the healing phase, consistent with immune homeostasis. Histologic healing in UC involves compartment-specific shifts in T lymphocyte populations and a marked reduction in IL-6 expression, reflecting coordinated immune regulation beyond clinical remission. These findings highlight the potential of combined cellular and cytokine biomarkers to monitor mucosal healing and guide immunomodulatory therapies.

Diagnosing urothelial carcinoma (UC) is usually a quite simple task but requires thoroughly examination of several slides; cases with more than 10 slides are not uncommon. Thus, an automated method for histopathological analysis is more than welcome. We selected from our archives 105 patients (100 UC and 5 cystitis); we examined the slides and selected and scanned one slide/case, obtaining whole slide images (WSIs). We performed a pixel-per-pixel semantic segmentation of 21 selected areas/WSI for several classes (high-/low-grade tumor, invasion, emboli, stroma, vessels, smooth muscle, etc.). We trained an InternImage model on this data set; we used dice coefficient (DCC) and intersection-over-union (IoU) as metrics for our model performance. UC patients were predominantly males (72%), average age 66.04years, 46% low-grade UC/ 54% high-grade UC, 42% noninvasive/ 58% invasive (28%pT1 and 30%pT2 or above). There were, on average, 3.93 paraffin blocks/case (1-17 paraffin blocks/case). The data set obtained after annotation was arbitrarily separated in training (57.18%), validation (21.37%) and test sets (21.44%). The results on test set are: high-grade tumor (0.66 DCC/0.49 IoU), low-grade tumors (0.82 DCC/0.70 IoU), stroma (0.84 DCC/0.73 IoU), vessels (0.75 DCC/0.60 IoU) and LVI (0.77 DCC/0.62 IoU). We evaluated each patch of the test set; apparently low DCC and IoU scores are consequences of human inability in precise drawing of the classes and/or impossibility of annotation of very small vessels. Our model identifies high-/low-grade tumor, invasion, emboli, and smooth muscle and highlights them on a heat map. The pathologist analyses highlighted areas, thus shortening the time required by microscopic analysis. The results of our model are encouraging; its use improves the diagnostic accuracy, reduces the time taken for analysis, and potentially leads to better patient outcomes.

Background: Histologic healing is increasingly recognized as a sensitive marker of disease remission in ulcerative colitis (UC). However, the dynamics of mucosal T lymphocytes and pro-inflammatory cytokines during healing remain incompletely understood. Methods: In this observational, cross-sectional study, paired colonic biopsies from 20 adult UC patients were analyzed during active inflammation and subsequent histologic healing. Immunohistochemistry was performed for CD3, CD4, CD8, and IL-6. Lymphocyte densities were quantified in intraepithelial and lamina propria compartments, while IL-6 expression was scored semi-quantitatively. Histologic activity was assessed using the Geboes score. Results: Intraepithelial CD4⁺ T cells significantly decreased during histologic healing (mean 6.8 → 3.75 cells/HPF, p < 0.05), whereas lamina propria CD4⁺ cells remained variably persistent, suggesting ongoing immune regulation. Intraepithelial CD8⁺ cells increased during remission, indicating a potential reparative or surveillance role. IL-6 expression markedly declined in epithelial and stromal compartments during healing, reflecting resolution of mucosal inflammation. Correlation analyses revealed enhanced coordination between CD4⁺ and CD8⁺ cells in the healing phase, consistent with immune homeostasis. Conclusions: Histologic healing in UC involves compartment-specific shifts in T lymphocyte populations and a marked reduction of IL-6 expression, reflecting coordinated immune regulation beyond clinical remission. These findings highlight the potential of combined cellular and cytokine biomarkers to monitor mucosal healing and guide immunomodulatory therapies.

A 67-year-old woman presented to the outpatient clinic with new-onset epigastric pain, with an initial abdominal ultrasound revealing a probable gastric mass. At endoscopy, we observed an approximately 5-cm subepithelial gastric tumor on the lesser curvature of the stomach and performed bite-on-bite biopsy, with a resulting histopathologic diagnosis of gastric leiomyoma being made ([Fig. 1]). We considered submucosal tunneling techniques for removal, but, owing to the large tumor size and its position, our team opted for laparoscopic and endoscopic cooperative surgery (LECS) [1] [2] for local tumor resection ([Fig. 2]; [Video 1]).

Eosinophilic fasciitis (EF) remains a diagnostic challenge due to its rarity and resemblance to scleroderma. This case report aims to provide a cohesive exploration of EF’s clinical nuances, emphasizing the importance of accurate diagnosis and effective management. A 52-year-old male developed bilateral forearm and calf hardening, along with erythema, pruritus, and pain four months prior to the presentation in our Clinic. The symptoms initially debuted bilaterally in the forearms and progressed to involve the calves, distal arms, and thighs. Clinical examination revealed symmetrical plaques on forearms and calves, featuring erythematous, hyper, and hypopigmented elements extending proximally, a positive “groove sign” and a moderate difficulty in knee joint flexion. Despite these findings, the patient was generally in good condition, without any other notable clinical signs. Initial laboratory findings showed slightly increased percentual eosinophil levels, elevated C-reactive protein (CRP), normal erythrocyte sedimentation rate (ESR), and negative antinuclear and scleroderma specific antibodies. Magnetic resonance imaging (MRI) demonstrated enhanced fascial signal and thickening while the fascia-muscle biopsy revealed marked edema and inflammatory lymphoplasmacytic infiltrate, consistent with the diagnosis of EF. The patient showed a favorable response to systemic corticosteroids. EF predominantly affects males aged 30 to 60 and is characterized by a sudden onset and unclear etiological factors. Differential diagnosis requires careful exclusion of scleroderma and other mimicking conditions. Diagnostic modalities such as skin-muscle biopsy and MRI reveal characteristic findings like inflammatory infiltrate and fascial thickening. Accurate diagnosis and differentiation from scleroderma are crucial, with early intervention involving glucocorticoids and immunosuppressive agents improving long-term outcomes.

The first lung transplant (LT) was made in Romania in 2018 at a 36-year-old male patient with chronic obstructive pulmonary disease (COPD). The study follows the first LT rehabilitation by describing the physical therapy program (PTP), the measurements of body mass and appendicular skeletal muscle mass (ASM) by bio-impedancemetry analysis (BIA) and the functional capacity assessment realized by the six-minute walk test (6MWT) and by the functional respiratory tests (FRTs) in order to evaluate the effectiveness of functional respiratory rehabilitation in this case during a period of one year. In parallel, repeated transbronchial biopsies were performed after six weeks, three months, six months and one year since the transplant. Only the first biopsies showed injuries suggesting an acute rejection, all the rest revealing mild, unspecific lesions. The patient followed 15 sessions of respiratory exercises, joints mobilizations and progressive global muscle strength started one month after LT surgery and was also instructed to perform the exercises at home, using a tablet given at discharge and under monthly guidance through telemedicine. All the measurements were performed before and after the rehabilitation cure, and it was repeated at three different evaluations for one year. The results showed that at the end of follow-up, the 6MWT was significantly increased from 59% of predicted distance at the intake in post-acute hospitalization to 166% at one year after LT, without desaturation that represent a very good evolution; the FRTs increased to normal, and the body weight increased with 18 kg (from severe underweight to normal weight) with constant increasement of skeletal muscle mass. The use of PTP after LT surgery significantly improves functional capacity and increases body mass and skeletal muscle mass.

Aims We aimed to assess the capacity of trainees with minimal NBI experience to classify gastric atrophy and intestinal metaplasia (IM) during routine upper gastrointestinal endoscopies performed with HD scopes.

In the present study, we employed the ddPCR and IHC techniques to assess the prevalence and roles of RAS and RAF mutations in a small batch of melanoma (n = 22), benign moles (n = 15), and normal skin samples (n = 15). Mutational screening revealed the coexistence of BRAF and NRAS mutations in melanomas and nevi and the occurrence of NRAS G12/G13 variants in healthy skin. All investigated nevi had driver mutations in the BRAF or NRAS genes and elevated p16 protein expression, indicating cell cycle arrest despite an increased mutational burden. BRAF V600 mutations were identified in 54% of melanomas, and NRAS G12/G13 mutations in 50%. The BRAF mutations were associated with the Breslow index (BI) (p = 0.029) and TIL infiltration (p = 0.027), whereas the NRAS mutations correlated with the BI (p = 0.01) and the mitotic index (p = 0.04). Here, we demonstrate that the “young” ddPCR technology is as effective as a CE-IVD marked real-time PCR method for detecting BRAF V600 hotspot mutations in tumor biopsies and recommend it for extended use in clinical settings. Moreover, ddPCR was able to detect low-frequency hotspot mutations, such as NRAS G12/G13, in our tissue specimens, which makes it a promising tool for investigating the mutational landscape of sun-damaged skin, benign nevi, and melanomas in more extensive clinical studies.

Cholangiocarcinoma (CCA) is a rare form of cancer, accounting for approximately 3% of all gastrointestinal cancers and 15% of primary liver malignancies. Most patients will require a multidisciplinary approach, including palliative, radiotherapy and systemic therapy (chemotherapy or, more recently, immunotherapy). Local ablation therapy using radiofrequency ablation (RFA) has also been explored in the setting of perihiliar cholangiocarcinoma (pCCA) as an adjunctive to palliative drainage. Tumour-infiltrating lymphocytes (TILs) are an adaptive immune cell type that can travel directly to the tumoural site to suppress cancer proliferation and development. Immune checkpoint inhibitor (ICI)-based therapies that act directly on the T-cell signalling pathway have proven particularly useful in recent trials, with molecules targeting the cytotoxic T-lymphocytes associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1)/programmed cell death ligand 1 (PD-L1) pathways showing efficacy in a wide range of indications. In this context, our study aimed to measure the potential effect of RFA on the intratumoural expression of checkpoint molecules, with an impact on modulating immunotherapy. We present here an exploratory study presenting an optimized flow cytometry protocol for tissue dissociation and cell characterization in terms of PD-1/PD-L1 and CTLA-4 expression. Our results show that RFA significantly increases the infiltration of cluster of differentiation 8(+ )(CD8(+)) cells and that the expression of PD-1 and CTLA-4 was significantly increased on cluster of differentiation 3(+) (CD3(+)) cells after RFA. RFA might promote a "switch" from an "immune-cold" to an "immune-hot" tumour microenvironment (TME) profile, with a significant impact on enabling immunotherapy for CCA patients.

Tuberculosis is the primary cause of death due to infection in the world. Identification of Mycobacterium tuberculosis in sputum is a diagnostic test, which can be used in screening programs—especially in countries with a high incidence of tuberculosis—to identify and treat those persons with the highest risk of disseminating the infection. We previously developed an algorithm which is able to automatically detect mycobacteria on tissue; in particular, our algorithm identified acid-fast bacilli on tissue with 100% specificity, 95.65% sensitivity, and 98.33% accuracy. We tested this algorithm on 1059 Ziehl–Neelsen-stained sputum smears to evaluate its results as a possible tool for screening. The results were displayed as a heat map of 32 × 32 pixel patches. Analysis of the positive patches revealed a good specificity (86.84%) and 100% sensitivity for patches with a level of confidence over 90; furthermore, the accuracy remained over 95% for all levels of confidence over 80, except the class (95–100]. The modest specificity is caused by the peculiarities of smears (uneven thickness, dust contamination, lack of coverslip). We will train the algorithm on sputum smears to increase the specificity to over 95%. However, as our algorithm showed no false negatives, it is suitable for screening.

BACKGROUND:Matrix metalloproteinase (MMP)1, MMP9, MMP11, and MMP13 are overexpressed in malignant melanoma (MM), being associated with tumor invasive phase, metastases, and more aggressive neoplastic phenotypes.AIM:The main objective of the current study was to correlate the expression of the MMPs with the evolution of MM toward distant metastasis.PATIENTS, MATERIALS AND METHODS:We designed a retrospective cohort study, including 13 patients with metastatic MM. Data concerning age, sex, localization of the primary lesion and metastasis, and histological and immunohistochemical features (intensity of expression and percent of positive cells for MMPs) were statistically processed.RESULTS:The time between the diagnosis of primitive melanoma and the diagnosis of metastasis ranged between 0 and 73 months, with a mean value of 18.3 months. The metastases rich in MMP1- and MMP9-positive cells occurred earlier than the metastases with low levels of positive cells. The mean period until metastasis was shorter for the MMP1-expressing tumors than the ones without MMP1 expression. MMP13 expression in the tumor and its metastasis was significantly linked with the time until the metastasis occurrence.CONCLUSIONS:This study emphasizes the roles of MMP1, MMP9, and MMP13 in the process of metastasis in melanoma and the opportunity to use them as therapeutic targets and surveillance molecules.