TACE inhibition: a promising therapeutic intervention against AATF-mediated steatohepatitis to hepatocarcinogenesis.

Metabolic dysfunction-associated steatohepatitis-driven hepatocellular carcinoma (MASH-HCC) is a global clinical challenge for which there is a limited understanding of disease pathogenesis and a subsequent lack of therapeutic interventions. We previously identified that tumor necrosis factor-alpha (TNF-α) upregulated apoptosis antagonizing transcription factor (AATF) in MASH. Here, we investigated the effect of TNF-α converting enzyme (TACE) inhibition as a promising targeted therapy against AATF-mediated steatohepatitis to hepatocarcinogenesis. A preclinical murine model that recapitulates human MASH-HCC was used in the study. C57Bl/6 mice were fed with chow diet normal water (CD) or western diet sugar water (WD) along with a low dose of carbon tetrachloride (CCl4; 0.2 µL·g-1, weekly) for 24 weeks. TACE activity, TNF-α levels, and AATF expression were measured. The mice were treated with the TACE inhibitor Marimastat for 12 weeks, followed by analyses of liver injury, fibrosis, inflammation, and oncogenic signaling. In vitro experiments using stable clones of AATF control and AATF knockdown were also conducted. We found that AATF expression was upregulated in WD/CCl4 mice, which developed severe MASH at 12 weeks and advanced fibrosis with HCC at 24 weeks. WD/CCl4 mice showed increased TACE activity with reduced hepatic expression of sirtuin 1 (Sirt1) and tissue inhibitor of metalloproteinase 3 (Timp3). The involvement of the SIRT1/TIMP3/TACE axis was confirmed by the release of TNF-α, which upregulated AATF, a key molecular driver of MASH-HCC. Interestingly, TACE inhibition by Marimastat reduced liver injury, dyslipidemia, AATF expression, and oncogenic signaling, effectively preventing hepatocarcinogenesis. Furthermore, Marimastat inhibited the activation of JNK, ERK1/2, and AKT, which are key regulators of tumorigenesis in WD/CCl4 mice and in AATF control cells, but had no effect on AATF knockdown cells. This study shows that TACE inhibition prevents AATF-mediated inflammation, fibrosis, and oncogenesis in MASH-HCC, offering a potential target for therapeutic intervention.

Glomerular parietal epithelial expression of CD44 in minimal change nephrotic syndrome and primary focal segmental glomerulosclerosis: A clinico-pathological study.

Introduction: Minimal change nephrotic syndrome (MCNS) and focal segmental glomerulosclerosis (FSGS) are the two common causes of nephrotic syndrome (NS) in both children and adults with overlapping clinical features, but with distinct prognostic and therapeutic implications. The distinction between these relies entirely on histopathology, which can sometimes be difficult. CD44 is expressed by activated parietal epithelial cells, plays a role in matrix deposition and thus in the pathogenesis of FSGS. Aims: To assess the expression of CD44 in MCNS and FSGS and to evaluate its association with the known clinical and histopathological prognostic factors. Materials and Methods: Thirty cases each of MCNS and FSGS were studied. The clinical, laboratory, histopathological, and CD 44 immunohistochemical data were recorded. The findings were analyzed and correlated. A P value of < 0.05 was considered statistically significant. Results: Statistical association was noted between CD44 positivity and serum creatinine (p = 0.031), estimated glomerular filtration rate (p = 0.040), segmental sclerosis (p < 0.001), tubular atrophy (p = 0.027), interstitial fibrosis (p = 0.027), and histological diagnosis (p < 0.001). The sensitivity, specificity, positive predictive, and negative predictive values were 90%, 76.67%, 79.41% and 88.46%, respectively. Conclusions: CD44 immunostain can effectively distinguish MCNS from FSGS. The congruent results of CD44 positivity with known prognostic factors support the possibility of using the CD44 marker as a predictive tool in selecting high-risk patients and offering appropriate therapeutic measures.

AATF inhibition exerts antiangiogenic effects against human hepatocellular carcinoma.

Background and aims: Angiogenesis is a key factor in the growth and metastasis of hepatic tumors and thus a potential therapeutic target in hepatocellular carcinoma (HCC). In this study, we aim to identify the key role of apoptosis antagonizing transcription factor (AATF) in tumor angiogenesis and its underlying mechanisms in HCC. Methods: HCC tissues were analyzed for AATF expression by qRT-PCR and immunohistochemistry. Stable clones of control and AATF knockdown (KD) were established in human HCC cells. The effect of AATF inhibition on the angiogenic processes was determined by proliferation, invasion, migration, chick chorioallantoic membrane (CAM) assay, zymography, and immunoblotting techniques. Results: We identified high levels of AATF in human HCC tissues compared to adjacent normal liver tissues, and the expression was found to be correlated with the stages and tumor grades of HCC. Inhibiting AATF in QGY-7703 cells resulted in higher levels of pigment epithelium-derived factor (PEDF) than controls due to decreased matric metalloproteinase activity. Conditioned media from AATF KD cells inhibited the proliferation, migration, and invasion of human umbilical vein endothelial cells as well as the vascularization of the chick chorioallantoic membrane. Furthermore, the VEGF-mediated downstream signaling pathway responsible for endothelial cell survival and vascular permeability, cell proliferation, and migration favoring angiogenesis was suppressed by AATF inhibition. Notably, PEDF inhibition effectively reversed the anti-angiogenic effect of AATF KD. Conclusion: Our study reports the first evidence that the therapeutic strategy based on the inhibition of AATF to disrupt tumor angiogenesis may serve as a promising approach for HCC treatment.

Pleomorphic adenoma of the external auditory canal.

Pleomorphic adenoma (PA) of the external auditory canal (EAC) is a rare clinical entity with a few cases reported in the literature. The clinical diagnosis of these lesions can be formidable due to their rarity and unusual location. This tumor occurs at various other anatomical locations apart from the major salivary glands. A 30-year-old female presented with a two-year history of a gradually enlarging and painless mass in the left external auditory canal. The tumor was excised, and histopathological and immunohistochemical evaluation revealed a mixed tumor with both epithelial and stromal components of different proportions, recognized and classified today by the World Health Organization (WHO) as a pleomorphic adenoma. The post-operative course was uneventful, and at the 10-month follow-up, no recurrence of the pleomorphic adenoma was noted. We highlight the histological features and the immunohistochemical profile of the tumor and review the literature on glandular neoplasms of the EAC and their recent classification, emphasizing on the histogenesis, clinical presentations, and microscopic features of the tumor. In addition, we aim to discuss vital features in differentiating these tumors from other tumors of the external auditory canal to enable clinicians and pathologists to recognize this uncommon benign neoplasm.

Unraveling the Potential Role of Tecomella undulata in Experimental NASH.

The pathophysiology of nonalcoholic steatohepatitis (NASH) is complex, owing to its diverse pathological drivers and, until recently, there were no approved drugs for this disease. Tecomella is a popular herbal medicine used to treat hepatosplenomegaly, hepatitis, and obesity. However, the potential role of Tecomella undulata in NASH has not yet been scientifically investigated. The administration of Tecomella undulata via oral gavage lowered body weight, insulin resistance, alanine transaminase (ALT), aspartate transaminase (AST), triglycerides, and total cholesterol in western diet sugar water (WDSW) fed mice but had no effect on chow diet normal water (CDNW) fed mice. Tecomella undulata improved steatosis, lobular inflammation, and hepatocyte ballooning and resolved NASH in WDSW mice. Furthermore, Tecomella undulata also alleviated the WDSW-induced Endoplasmic Reticulum stress and oxidative stress, enhanced antioxidant status, and thus reduced inflammation in the treated mice. Of note, these effects were comparable to saroglitazar, the approved drug used to treat human NASH and the positive control used in the study. Thus, our findings indicate the potential of Tecomella undulata to ameliorate WDSW-induced steatohepatitis, and these preclinical data provide a strong rationale for assessing Tecomella undulata for the treatment of NASH.

Pleomorphic adenoma of the external auditory canal

ABSTRACT Pleomorphic adenoma (PA) of the external auditory canal (EAC) is a rare clinical entity with a few cases reported in the literature. The clinical diagnosis of these lesions can be formidable due to their rarity and unusual location. This tumor occurs at various other anatomical locations apart from the major salivary glands. A 30-year-old female presented with a two-year history of a gradually enlarging and painless mass in the left external auditory canal. The tumor was excised, and histopathological and immunohistochemical evaluation revealed a mixed tumor with both epithelial and stromal components of different proportions, recognized and classified today by the World Health Organization (WHO) as a pleomorphic adenoma. The post-operative course was uneventful, and at the 10-month follow-up, no recurrence of the pleomorphic adenoma was noted. We highlight the histological features and the immunohistochemical profile of the tumor and review the literature on glandular neoplasms of the EAC and their recent classification, emphasizing on the histogenesis, clinical presentations, and microscopic features of the tumor. In addition, we aim to discuss vital features in differentiating these tumors from other tumors of the external auditory canal to enable clinicians and pathologists to recognize this uncommon benign neoplasm.

A clinico-pathological study of lupus nephritis based on the International Society of Nephrology-Renal Pathology Society 2003 classification system.

INTRODUCTION: Lupus nephritis (LN) is a major complication of systemic lupus erythematosus (SLE). Renal involvement is a major determinant of the prognosis of SLE. The histological classification of LN is a key factor in determining the renal survival of patients with LN. Prompt recognition and treatment of renal disease are important, as early response to therapy is correlated with better outcome and renal biopsy plays an important role in achieving this. OBJECTIVES: The objective of this study was to correlate the clinical and laboratory findings with histopathological classes of LN as per the 2003 International Society of Nephrology-Renal Pathology Society (ISN/RPS) classification system. PATIENTS AND METHODS: Fifty-six patients with SLE, undergoing a renal biopsy for renal dysfunction were studied. The comparison of data from multiple groups was made by Pearson's Chi-square test and between two groups by independent samples t-test. The values of P < 0.05 were considered statistically significant. RESULTS: Of the 56 cases studied, 51 (91.1%) were females. The most common presenting symptoms were edema, arthralgia, and hypertension. Class IV (55.4%) was the most common class. Thirty-nine (69.6%) cases showed full house immunostaining. Hypertension, hematuria, proteinuria, and tubulo-interstitial disease showed a significant correlation (P < 0.05) with ISN/RPS classification, 2003. CONCLUSION: Assessment and management of patients with suspected LN are greatly facilitated through information obtained by renal biopsy. Since renal morphology may predict long-term prognosis, and no clinical or laboratory feature uniformly predicts prognosis, it is important to study the constellation of features in LN for better patient management.

Ki-67 immunostaining in astrocytomas: Association with histopathological grade - A South Indian study.

BACKGROUND: Astrocytomas are the most common primary tumor of the central nervous system. The distinction between different tumor grades can be tested despite criteria given by the World Health Organization (WHO). Ki-67 is a potent biological marker used in grading of astrocytomas, which estimates growth of the neoplasm quantitatively and will help in predicting prognosis accurately. OBJECTIVES: The aim of this was to study the proliferative activity using Ki-67 immunostaining and to assess the relationship of Ki-67 staining with the histopathological grading of astrocytomas. PATIENTS AND METHODS: Thirty cases of histologically proven astrocytomas were studied. The histopathological grade was assessed using the 2007 WHO criteria. Immunohistochemistry for Ki-67 was done on paraffin-embedded wax sections. P < 0.05 was considered statistically significant. RESULTS: Thirty cases of astrocytomas studied showed a male preponderance (M:F = 1.72:1) with a mean age of 48.1 years. Of these, Grade I, (n = 1, 3.33%), Grade II, (n = 7, 23.3%), Grade III (n = 6, 20%), and Grade IV (n = 16, 53.3%) astrocytomas were analyzed. The mean Ki-67 labeling index (LI) in Grades I, II, III, and IV was 0.02, 0.81, 9.14, and 17.81, respectively. Statistically significant difference was seen in the Ki-67 LI of low-grade (Grade II) and high-grade astrocytomas (Grades III and IV). There was concordance between histopathological grading and Ki-67 LI in 27 (90%) and discordance in 3 (10%) cases. CONCLUSION: Ki-67 LI varies considerably in different grades of astrocytomas and considerable overlaps can be observed between them. It can be of great help in situations where there is a lack of correlation between clinical parameters and histopathological diagnosis. Determination of Ki-67 LI should constitute a part of routine investigations in patients with astrocytomas.

Comparison of Cell Preparations between Commercially Available Filter Cards of the Cytospin with Custom Made Filter Cards.

INTRODUCTION: Cytological evaluation of body fluids is an important diagnostic technique. Cytocentrifuge has contributed immensely to improve the diagnostic yield of the body fluids. Cytocentrifuge requires a filter card for absorbing the cell free fluid. This is the only consumable which needs to be purchased from the manufacturer at a significant cost. AIM: To compare the cell density in cytocentrifuge preparations made from commercially available filter cards with custom made filter cards. MATERIALS AND METHODS: This was a prospective analytical study undertaken in department of pathology of a tertiary care centre. A 300 GSM handmade paper with the absorbability similar to the conventional card was obtained and fashioned to suit the filter card slot of the cytospin. Thirty seven body fluids were centrifuged using both conventional and custom made filter card. The cell density was measured as number of cells per 10 high power fields. The median cell density was compared using Mann-Whitney U test. The agreement between the values was analysed using Bland Altman analysis. RESULTS: The median cell count per 10 High power field (HPF) with conventional card was 386 and that with custom made card was 408. The difference was not statistically significant (p = 0.66). There was no significant difference in the cell density and alteration in the morphology between the cell preparations using both the cards. CONCLUSION: Custom made filter card can be used for cytospin cell preparations of body fluids without loss of cell density or alteration in the cell morphology and at a very low cost.

Cannonballs in Pap Smears: Double Whammy of Bacterial Vaginosis and Associated Infections.

INTRODUCTION: Cannonballs are structures consisting of neutrophils adherent to epithelial cells, often seen in vaginal Pap smears of patients with trichomoniasis and chlamydiasis. We frequently observed these cannonballs in inflammatory Pap smears showing evidence of bacterial vaginosis. OBJECTIVE: To study the association between cannonballs and bacterial vaginosis with associated infections in inflammatory Pap smears. STUDY DESIGN: This was a retrospective study performed over a period of 6 months between April 2014 and September 2014. Three hundred consecutive Pap smears assessed during the study period were retrieved from the archives and studied. RESULTS: A total of 280 smears were studied. Neutrophilic infiltrates were seen in 83.6%, cannonballs in 82.3%, and bacterial vaginosis in 70.7% of the smears. Cannonballs were found in 84.4% of the smears showing evidence of bacterial vaginosis with associated inflammatory infiltrates. There was a significant association between cannonballs and bacterial vaginosis (p = 0.0001). The odds ratio was 13.8 (95% CI: 7.2-26.2). CONCLUSION: The present study shows a significant association between cannonballs and bacterial vaginosis and associated vaginal infections.

Myxoinflammatory fibroblastic sarcoma of the thigh: A morphologic diversity.

Myxoinflammatory fibroblastic sarcoma (MIFS)/acralmyxoinflammatory fibroblastic sarcoma (AMFS) is a rare, painless, low-grade neoplasm which commonly occurs in the extremities. It has a distinctive morphology and can be a diagnostic challenge, simulating inflammatory conditions as well as neoplastic conditions. They are low-grade sarcomas with a protracted clinical course, a high rate of local recurrence and a low rate of metastasis. We report a case of proximal MIFS in a 50-year-old woman who presented with a mass in the thigh.

Fine needle aspiration cytology of pseudosarcomatous reactive lesions of soft tissues: A report of two cases.

Pseudosarcomatous lesions are reactive proliferative lesions of the soft tissue, that are likely to be misdiagnosed as malignant, based on clinical and histological features. The most common lesions are nodular fasciitis, proliferative fasciitis, proliferative myositis and myositis ossificans. These rapidly growing soft-tissue lesions can represent a variety of diagnoses involving radically different treatment modalities. Accurate diagnosis is important to avoid unnecessary and often mutilating surgery. We report two cases to illustrate the importance of correct identification of these lesions by fine needle aspiration cytology.