Search Results (59)

Hepatocyte apoptotic bodies (ApopBDs) are extracellular vesicles formed during hepatocyte apoptosis. Although they were initially recognized as cellular waste and vesicles that clear toxic substances and viral infections in the liver, they are now known to serve as key mediators of intercellular communication that influence key metabolic and immune responses, such as inflammation, regeneration, and fibrosis. While numerous functions of ApopBDs in the liver are emerging, this review will focus on discussing their biogenesis, characterization, and roles in different liver diseases, with an emphasis on intercellular communication with liver-resident cells. The mechanisms of liver injury are convoluted by series of injurious crosstalk between hepatocyte ApopBDs and surviving resident cells. A unique feature of liver injury is a constant cycle of hepatocyte apoptosis, which has been attributed to crosstalk between surviving hepatocytes and their ApopBDs. The progression of liver injury is also affected by the activation of proinflammatory and profibrotic pathways such as TLR9/NLRP3 and JAK-STAT3. Given the expression of hepatocyte-specific molecular signatures on these ApopBDs, their application as diagnostic tools may improve the treatment of liver diseases. Although the science of hepatocyte ApopBDs is fairly recent and still emerging, in-depth understanding of this aspect of liver biology may provide a novel therapeutic option for the progression of liver damage. Full article

Chronic HBV infection remains a global health challenge, driving liver fibrosis, cirrhosis, and hepatocellular carcinoma (HCC). Liver injury is primarily mediated by host immune responses rather than direct viral cytotoxicity. Macrophages, including Kupffer cells, play dual roles in antiviral defense and disease progression. HBV skews macrophages toward an M2-like, immunosuppressive phenotype, promoting viral persistence and fibrogenesis via cytokines such as Interleukin (IL)-10 and Transforming growth factor-beta (TGF-β). Therapeutic strategies targeting macrophage polarization, including Toll-like receptor (TLR) agonists, immune checkpoint inhibitors, and nanoparticle-based systems, are under investigation. Addressing macrophage heterogeneity and the immunosuppressive hepatic microenvironment using advanced models is essential. Modulating macrophages offers a promising avenue to control HBV, restore immune balance, and mitigate liver injury. This review highlights the central role of macrophages in chronic HBV infection and explores emerging therapeutic strategies. Full article

Aging is a critical factor influencing susceptibility to hepatic injury. In this study, the spontaneous development of liver injury with advancing age and potential sex-related differences in these processes are examined. This study focuses on key mechanisms such as fatty acid metabolism, immune response, and cellular stress in male and female C57BL/6 mice. Aged male and female mice (20 to 22 months old) exhibited higher body weight and an altered metabolic profile and fatty acid metabolism compared to their younger counterparts (8 to 10 weeks old). In addition, increased oxidative stress, cellular senescence, expression of inflammatory markers, and cytokines/chemokines levels were also observed in aged male and female mice compared to younger mice. Furthermore, the aged mice exhibited increased indices of hepatic fibrosis, evident from the upregulation of smooth muscle actin-α, collagen, and transforming growth factor-β. In conclusion, aging promotes spontaneous liver injury by increasing indices of oxidative stress, steatosis, inflammation, and fibrosis. These results highlight the impact of chronological age on the liver that can increase its susceptibility to secondary hepatic stressors such as alcohol, high-calorie diet, or hepatotropic infections. Understanding how metabolic and inflammatory pathways change with aging in males and females is essential for elucidating the mechanisms that drive chronic liver disease progression. These insights are particularly important for developing targeted, sex-specific prevention and therapeutic strategies for the aging population. Full article

Background/Objectives: This study aims to critically appraise the role of para-aortic surgical staging in locally advanced cervical cancer (LACC) in the era of advanced imaging, and to outline how selective surgery and biomarkers could be integrated within modern, quality-assured treatment pathways. Methods: Narrative review of randomized trials, large databases, and prospective/retrospective series comparing para-aortic lymphadenectomy with imaging-based staging; focused appraisal of Uterus-11, NCDB analyses, and ongoing prospective trials (PAROLA with Senti-PAROLA as one of its sub-studies and PALDISC). Emerging technologies (PET/MRI, radiomics/AI) and molecular assays (OSNA, HPV-ctDNA) were also assessed. Results: PET/CT remains the standard for distant staging, but sensitivity for low-volume nodal disease (<5 mm) is poor; in pelvic-positive/para-aortic-negative patients, occult para-aortic metastases approach ~21%. Para-aortic surgical staging modifies radiotherapy planning in ~18% of cases and can act as a de-escalation tool by avoiding unnecessary extended-field CRT (EF-CRT) when para-aortic nodes are negative. Uterus-11 showed no overall survival difference versus CT-based staging, but suggested benefit in FIGO 2009 stage IIB; its design (CT comparator, optimistic assumptions, limited power) constrains inference. Minimally invasive extraperitoneal/transperitoneal staging is feasible with low morbidity in expert centers, yet real-world management may worsen outcomes. The role of systemic intensification in node-positive disease remains undefined: PALN-positive patients were excluded from the INTERLACE trial. In the KEYNOTE-826 study, subgroup analyses according to nodal status were not reported, although the benefit of pembrolizumab remained consistent irrespective of bevacizumab use. Sentinel para-aortic mapping and biomarkers (e.g., HPV-ctDNA) may refine selection and reduce morbidity. Conclusions: Surgical staging is the most accurate method to detect occult para-aortic disease. Its routine use is not justified, but it may benefit selected high-risk patients, particularly where decisions on EF-CRT or systemic therapy hinge on para-aortic status. Future practice should integrate advanced imaging, selective surgery, and biomarkers within accredited centers, guided by large collaborative trials conducted under international quality frameworks such as ESGO/ESTRO/ESP guidelines. Full article

Background and Objectives: The aim of this prospective study was to evaluate the diagnostic accuracy of the one-step nucleic acid amplification (OSNA) assay for the intraoperative assessment of sentinel lymph node (SN) metastases, including micrometastases in patients with stage IA low-grade endometrial cancer. Materials and Methods: A prospective analysis was conducted on 204 patients with low-risk endometrial cancer who underwent hysterectomy, bilateral salpingo-oophorectomy, and sentinel node navigation surgery. SNs were analyzed intraoperatively using the OSNA assay, and positive patients underwent systematic pelvic lymphadenectomy. Results: Among the 204 patients included, SN metastases were identified in 12 patients (6%), including 10 patients with micrometastases and 2 patients with macrometastases. No metastases were detected in non-SNs in any of the 12 patients. Recurrence occurred in two patients (1%), involving the vaginal stump and pelvic cavity dissemination, but no lymph node recurrence was observed. The OSNA assay identified a proportion of micrometastases in low-risk endometrial cancer. While a direct comparison with conventional pathological ultra-staging was not performed in this study, the detection rate of micrometastases appears higher than that reported in historical controls. Conclusions: This is the first prospective study to evaluate the intraoperative use of the OSNA assay for whole SNs in endometrial cancer. The results suggest that the OSNA assay enhances the detection of micrometastases, enabling a more accurate assessment of SN metastases. In low-risk endometrial cancer, systematic pelvic lymphadenectomy may be safely omitted in patients with SN-positive micrometastases. Further prospective studies are necessary to validate these findings and support the adoption of this approach in clinical practice. Full article

Background/Objectives: Lymph node metastases (LNM) undetected by standard hematoxylin and eosin (H&E) have been associated with unfavorable prognosis in colorectal cancer. The One-Step Nucleic Acid Amplification (OSNA) assay has demonstrated superior sensitivity in detecting LNM compared to H&E. We aimed to assess the performance of OSNA in detecting LNM, as well as its prognostic value in rectal cancer (RC) patients. Methods: Lymph nodes (LNs) of patients from 15 centers were analyzed by both H&E and OSNA. The total tumor load (TTL) was defined as the sum of cytokeratin 19 mRNA copies/µL in all LNs from a surgical specimen, using a threshold of 250 copies/μL for OSNA positivity. Cox proportional hazard regression was used to assess the effect of TTL ≥ 250 or 6000 copies/μL on cancer-specific survival (CSS) and recurrence-free survival (RFS), with Firth’s method applied to account for low event rate. Results: A total of 97 RC patients were included. Of these, 84 patients were eligible for survival analysis. The sensitivity and specificity of OSNA, compared to H&E, were 91.7% and 84.7%, respectively. TTL ≥ 6000 versus <6000 copies/μL was related to worse CSS and RFS. When dividing TTL into three groups: ≤250, 250–6000, and >6000 copies/μL, only TTL ≥ 6000 copies/μL was significantly associated with worse CSS and RFS. Conclusions: The OSNA assay is highly sensitive for detecting LNM in RC patients. A TTL of ≥6000 copies/μL could identify a subset of RC patients with worse CSS and RFS who might benefit from adjuvant treatment or intensive surveillance. Full article

Objective: This study aimed to evaluate the reliability and clinical utility of intraoperative sentinel lymph node (SN) metastasis diagnosis using the one-step nucleic acid amplification (OSNA) assay in early-stage cervical cancer by comparing its accuracy with conventional histopathological examination. Methods: A retrospective analysis was conducted on 163 patients who underwent SN biopsy at Kagoshima University Hospital between April 2014 and December 2024. This study included 50 and 113 patients in the OSNA assay and histopathological diagnosis groups, respectively. The OSNA assay quantified cytokeratin 19 (CK19) mRNA levels to determine SN metastasis. The surgical outcomes, SN metastasis detection rates, and non-SN metastasis status were compared between the two diagnostic methods. Results: The SN metastasis detection rate was significantly higher in the OSNA group (12%) than in the pathology group (3%) (p < 0.05). The OSNA assay identified only micrometastases (+) among the positive cases, whereas histopathological diagnosis detected both macrometastases and micrometastases. No non-SN metastases were observed in any of the SN-positive cases, and no significant differences were observed in the recurrence rates between the two groups. Conclusions: The OSNA assay demonstrated a higher SN metastasis detection rate than conventional pathology and demonstrated superior sensitivity in identifying micrometastases. These findings suggest that intraoperative OSNA-based SN assessment in cervical cancer could improve staging accuracy and potentially reduce the need for systematic lymphadenectomy. However, further prospective studies are warranted to confirm these findings and establish clinical guidelines. Full article

Background: Current international guidelines recommend omitting axillary lymph node dissection (ALND) based on sentinel lymph node biopsy (SLNB) in early-stage breast cancer patients. However, the evolving landscape of axillary management highlights the need to balance diagnostic accuracy with minimizing invasiveness. The possibility of omitting SLNB itself should also be considered. Methods: In this study, we have evaluated the feasibility of omitting SLNB in a total of 1044 clinically node-negative (cN (−)) breast cancer patients whose SLN status was determined by histopathology and one-step nucleic acid amplification (OSNA) after SLNB. We also analyzed SLN-positive cases to explore the association between non-SLN (NSLN) metastatic status and various biomarkers. We predicted the metastatic status of NSLNs based on patient data using a nomogram and further assessed the prevalence of macro- and micro-metastatic SLN, along with the NSLN status in SLNB cases. Results: Of the 644 cN (−) cases, approximately 70% of SLN-positive cases were NSLN negative, suggesting that ALND could be omitted. SLN (+) was detected approximately 7% more often by OSNA than by histopathology, suggesting that OSNA detection may be an overdiagnosis. Although NSLN-positive cases represented only 5.9% of the 581 cN (−) cases and, therefore, ALND could be omitted, it may be difficult to omit the SLNB itself as the SLN macro-metastasis was 12.5%. Biomarker analysis showed a significant correlation between total tumor load and metastatic SLN copy number with NSLN metastatic status. Based on these tumor characteristics, the nomogram predicted NSLN-positive rates very well. Conclusions: Thus, omitting SLNB itself carries the risk of missing high-frequency macro-metastatic SLN-positive cases and losing important SLN-related information that can predict NSLN metastases. Therefore, SLNB, which provides not only SLN status but also NSLN metastases, is necessary for reassurance in omitting ALND. Full article

Breast cancer is one of the most common cancers diagnosed in both countries with high and low levels of socio-academic development. Routine, regular screening tests being introduced in an increasing number of countries make it possible to detect breast cancer at an early stage of development, as a result of which the trend in the incidence of metastatic breast cancer has been decreasing in recent years. The latest guidelines for the treatment of this tumor do not recommend axillary dissection, which limits the need for rapid assessment of the nodes during surgery. Regardless of the progression of the disease, lymph node biopsy and their analysis is one of the most common diagnostic methods for detecting metastases. Systems using one-step amplification of nucleic acids have been present in the diagnosis of breast cancer for nearly 20 years. The one-step nucleic acid amplification (OSNA) test semi-quantitatively detects the number of cytokeratin 19 mRNA copies, a well-known tumor marker, which can be used to infer the presence of metastases in non-sentinel lymph nodes (SLN). Aim: OSNA is a widely used molecular method for SLN, intra-, or postoperative analysis. Its high accuracy has been proved over the years in clinical use. In this review, we checked current state of this technology and compared it to its competitors in the field of breast cancer diagnosis in the era of Axillary Lymph Nodes Dissection (ALND) importance decrease with intention to foresee its further potential use. Objectives: To evaluate OSNA current place in breast cancer diagnosis and treatment we compared OSNA to other lymph node assessing methods. We based our review on original articles and metanalyses published in the last decade. The research was conducted with PubMed, Science Direct, Google Scholar, and NCBI databases. The collected data allowed us to assess the accuracy of OSNA, its cost effectiveness, and its application in other cancers. Results: Regardless of the progression of the disease, a lymph node biopsy and its analysis constitutes one of the most common diagnostic methods for detecting metastases. The OSNA method is characterized by high sensitivity and specificity, and its predictive value has been confirmed by many studies over the years. While its cost effectiveness is still a matter of discussion, this method has been tested more thoroughly than other new lymph nodes assessing technologies. Conclusions: Despite the emergence of competing methods, this test is still widely used as a routine intraoperative examination of lymph nodes. Research carried out in recent years has proved its effectiveness in the diagnosis of other cancers, in the research field, and as a provider of additional data for prognosis improvement. Full article

About 296 million people worldwide are living with chronic hepatitis B viral (HBV) infection, and outcomes to end-stage liver diseases are potentiated by alcohol. HBV replicates in hepatocytes, but other liver non-parenchymal cells can sense the virus. In this study, we aimed to investigate the regulatory effects of macrophages on HBV marker and interferon-stimulated genes (ISGs) expressions in hepatocytes. This study was performed on HBV-replicating HepG2.2.15 cells and human monocyte-derived macrophages (MDMs). We found that exposure of HepG2.2.15 cells to an acetaldehyde-generating system (AGS) increased HBV RNA, HBV DNA, and cccDNA expressions and suppressed the activation of ISGs, APOBEC3G, ISG15, and OAS1. Supernatants collected from IFNα-activated MDMs decreased HBV marker levels and induced ISG activation in AGS-treated and untreated HepG2.215 cells. These effects were reversed by exposure of MDMs to ethanol and mimicked by treatment with exosome release inhibitor GW4869. We conclude that exosome-mediated crosstalk between IFN-activated macrophages and HBV-replicating hepatocytes plays a protective role via the up-regulation of ISGs and suppression of HBV replication. However, ethanol exposure to macrophages breaks this protection. Full article

Pelvic lymph node dissection (PLND) is the most accurate procedure for lymph node (LN) staging in prostate cancer (PCa) patients. LN sectioning and hematoxylin and eosin (H&E) staining of at least one slice remains the gold standard for LN evaluation, potentially leading to misdetection of small metastatic focus. Entire LN analysis is possible with One-Step Nucleic Acid Amplification (OSNA) by detecting cytokeratin 19 (CK19) mRNA as a surrogate for LN invasion. This study aimed to compare postoperative performance of OSNA pooling with conventional H&E staining for pathological LN detection in PCa patients. POPCORN was an observational, prospective, and multicenter study of patients with PCa who underwent PLND. Dissected LNs were analyzed by both methods. This study included 2503 LNs from 131 patients, showing no statistically significant differences in pathological LN detection. Concordance between methods was high (93.9%), as were specificity (96.6%) and negative predictive value (96.6%) of OSNA pooling. The measure of agreement (Cohen’s Kappa [κ]) was 0.70. Only eight (6.1%) discordances were observed, including four misdetections from each method. Results showed a high concordance between OSNA pooling and H&E staining, suggesting that OSNA pooling may be a good alternative to H&E staining to detect LN metastases in PCa patients. Full article

In the management of early-stage breast cancer (BC), lymph nodes (LNs) are typically characterised using the One-Step Nucleic Acid Amplification (OSNA) assay, a standard procedure for assessing subclinical metastasis in sentinel LNs (SLNs). The pivotal role of LNs in coordinating the immune response against BC is often overlooked. Our aim was to improve prognostic information provided by the OSNA assay and explore immune-related gene signatures in SLNs. The expression of an immune gene panel was analysed in SLNs from 32 patients with Luminal A early-stage BC (cT1-T2 N0). Using an unsupervised approach based on these expression values, this study identified two clusters, regardless of the SLN invasion: one evidencing an adaptive anti-tumoral immune response, characterised by an increase in naive B cells, follicular T helper cells, and activated NK cells; and another with a more undifferentiated response, with an increase in the activated-to-resting dendritic cells (DCs) ratio. Through a protein—protein interaction (PPI) network, we identified seven immunoregulatory hub genes: CD80, CD40, TNF, FCGR3A, CD163, FCGR3B, and CCR2. This study shows that, in Luminal A early-stage BC, SLNs gene expression studies enable the identification of distinct immune profiles that may influence prognosis stratification and highlight key genes that could serve as potential targets for immunotherapy. Full article

Lymphadenectomy represents a fundamental step in the staging and treatment of non-small cell lung cancer (NSCLC). To date, the extension of lymphadenectomy in early-stage NSCLC is a debated topic due to its possible complications. The detection of sentinel lymph nodes (SLNs) is a strategy that can improve the selection of patients in which a more extended lymphadenectomy is necessary. This pilot study aimed to refine lymph nodal staging in early-stage NSCLC patients who underwent robotic lung resection through the application of innovative intraoperative sentinel lymph node (SLN) identification and the pathological evaluation using one-step nucleic acid amplification (OSNA). Clinical N0 NSCLC patients planning to undergo robotic lung resection were selected. The day before surgery, all patients underwent radionuclide computed tomography (CT)-guided marking of the primary lung lesion and subsequently Single Photon Emission Computed Tomography (SPECT) to identify tracer migration and, consequently, the area with higher radioactivity. On the day of surgery, the lymph nodal radioactivity was detected intraoperatively using a gamma camera. SLN was defined as the lymph node with the highest numerical value of radioactivity. The OSNA amplification, detecting the mRNA of CK19, was used for the detection of nodal metastases in the lymph nodes, including SLN. From March to July 2021, a total of 8 patients (3 female; 5 male), with a mean age of 66 years (range 48–77), were enrolled in the study. No complications relating to the CT-guided marking or preoperative SPECT were found. An average of 5.3 lymph nodal stations were examined (range 2–8). N2 positivity was found in 3 out of 8 patients (37.5%). Consequently, pathological examination of lymph nodes with OSNA resulted in three upstages from the clinical IB stage to pathological IIIA stage. Moreover, in 1 patient (18%) with nodal upstaging, a positive node was intraoperatively identified as SLN. Comparing this protocol to the usual practice, no difference was found in terms of the operating time, conversion rate, and complication rate. Our preliminary experience suggests that sentinel lymph node detection, in association with the accurate pathological staging of cN0 patients achieved using OSNA, is safe and effective in the identification of metastasis, which is usually undetected by standard diagnostic methods. Full article

Alcohol-associated liver disease (ALD) is a substantial cause of morbidity and mortality worldwide and represents a spectrum of liver injury beginning with hepatic steatosis (fatty liver) progressing to inflammation and culminating in cirrhosis. Multiple factors contribute to ALD progression and disease severity. Here, we overview several crucial mechanisms related to ALD end-stage outcome development, such as epigenetic changes, cell death, hemolysis, hepatic stellate cells activation, and hepatic fatty acid binding protein 4. Additionally, in this review, we also present two clinically relevant models using human precision-cut liver slices and hepatic organoids to examine ALD pathogenesis and progression. Full article

Background and Objectives: This review paper highlights the key alternatives to the blue dye/radioisotope method of sentinel lymph node biopsy (SLNB). It analyses the research available on these alternative methods and their outcomes compared to the traditional techniques. Materials and Methods: This review focused on fifteen articles, of which five used indocyanine green (ICG) as a tracer, four used magnetic tracers, one used one-step nucleic acid amplification (OSNA) and Metasin (quantitative reverse transcriptase-polymerase chain reaction), one used the photosensitiser talaporfin sodium, one used sulphur hexafluoride gas microbubbles, one used CT-guided lymphography and two focused on general SLNB technique reviews. Results: Of the 15 papers analysed, the sentinel node detection rates were 69–100% for indocyanine green, 91.67–100% for magnetic tracers, 81% for talaporfin sodium, 9.3–55.2% for sulphur hexafluoride gas microbubbles, 90.5% for CTLG and 82.7–100% for one-step nucleic acid amplification. Conclusions: Indocyanine green fluorescence (ICG) and magnetic tracers have been proven non-inferior to traditional blue dye and isotope regarding SLNB localisation. Further studies are needed to investigate the use of these techniques in conjunction with each other and the possible use of language learning models. Dedicated studies are required to assess cost efficacy and longer-term outcomes. Full article