Synchronous hepatocellular carcinoma and renal cell carcinoma: A cytological marvel.

Multiple primary synchronous tumours have always created an inquisitiveness among clinicians, radiologists and pathologists. The diagnosis invariably proposes a challenge to diagnosticians. The coexistence of a primary hepatocellular carcinoma (HCC) and renal cell carcinoma (RCC) is exceedingly rare, with countable number of cases being reported in literature. We report a pioneer case of 75-year-old male, having chronic hepatitis B, diagnosed with synchronous primary RCC and HCC in by fine-needle aspiration cytology (FNAC) and confirmed by immunohistochemistry.

The Role of Novel Tiered Reporting System in Serous Fluid Cytology and Risk of Malignancy Assessment: A Retrospective Study in a Tertiary Care Center.

Background: Serous effusion cytology (SEC) reporting is important for the management of the cancer patient. The International System for Reporting Serous Fluid Cytology (ISRSFC) provides tiered reporting terminology to standardize practice, looking into the risk of malignancy (MAL) for each category. In this study, we have assessed the utility of the ISRSFC and reported our experience at a tertiary cancer center. Materials and Methods: Serous fluid cytology reported from January 2019 to December 2020 was categorized according to ISRSFC diagnostic categories: nondiagnostic (ND), negative for MAL (NFM), atypia of undetermined significance (AUS), suspicious for MAL (SFM), and MAL. The risk of MAL (ROM) and performance parameters were calculated. Results: A total of 2150 serous effusions including 1160 pleural, 929 peritoneal, and 61 pericardial effusions from 2071 patients were reported. There were 742 males and 1329 females. The patient's age ranged from less than 1 yr to 95 years. The volume of the sample ranged from 0.5 ml to 2000 ml. There were 114 ND (5.32%), 1068 NFM (49.67%), 144 AUS (6.69%), 82 SFM (3.81%), and 742 MAL (34.51%) cases. Adenocarcinoma was the most common MAL involving serous fluids (91.50%). The calculated ROM was 15.38% for ND, 24.26% for NFM, 62.96% for AUS, 79.16% for SFM, and 100% for MAL. Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy were, respectively, 68.04%, 96.96%, 96.35%, 72.07%, and 81.33%. Conclusion: ISRSFC provides consistent reporting terminology and the ROM for each category aids in clinical correlation and management.

Neuroblastoma in uncommon age group - A case series diagnosed on cytology.

BACKGROUND: Neuroblastoma is the most common extracranial malignant neoplasm in early childhood. It is rare in the adult population. AIMS AND OBJECTIVES: We aimed to study the incidence of neuroblastoma in the uncommon age group diagnosed on cytology. MATERIALS AND METHODS: A prospective descriptive study spanning 2 years from December 2020 to January 2022 was done, in which neuroblastoma cases diagnosed by Fine needle aspiration cytology aged >12 years were collected. The clinical, cytomorphological and immunohistochemical findings were studied. Histopathological correlation was done wherever available. RESULTS: We identified three cases of neuroblastoma during this period. Two cases were middle-aged adults, and one was an adolescent. All cases presented with abdominal masses and revealed small round cell tumor on cytology. Two cases fell into undifferentiated category and one case fell into the poorly differentiated subtype. All cases were positive for neuroendocrine markers. Histopathological correlation was available in two cases. MYC N amplification was absent in all cases. CONCLUSION: It differs from pediatric neuroblastoma due to the lack of classical histomorphological features and molecular alterations. Adult-onset neuroblastomas carry a worse prognosis than childhood tumors.

Mapping Immune-Tumor Bidirectional Dialogue Using Ultrasensitive Nanosensors for Accurate Diagnosis of Lung Cancer.

Lung cancer is one of the most common cancers with high mortality worldwide despite the development of molecularly targeted therapies and immunotherapies. A significant challenge in managing lung cancer is the accurate diagnosis of cancerous lesions owing to the lack of sensitive and specific biomarkers. The current procedure necessitates an invasive tissue biopsy for diagnosis and molecular subtyping, which presents patients with risk, morbidity, anxiety, and high false-positive rates. The high-risk diagnostic approach has highlighted the need to search for a reliable, low-risk noninvasive diagnostic approach to capture lung cancer heterogeneity precisely. The immune interaction profile of lung cancer is driven by immune cells' distinctive, precise interactions with the tumor microenvironment. Here, we hypothesize that immune cells, particularly T cells, can be used for accurate lung cancer diagnosis by exploiting the distinctive immune-tumor interaction by detecting the immune-diagnostic signature. We have developed an ultrasensitive T-sense nanosensor to probe these specific diagnostic signatures using the physical synthesis process of multiphoton ionization. Our research employed predictive in vitro models of lung cancers, cancer-associated T cells (PCAT, MCAT) and CSC-associated T cells (PCSCAT, MCSCAT), from primary and metastatic lung cancer patients to reveal the immune-diagnostic signature and uncover the molecular, functional, and phenotypic separation between patient-derived T cells (PDT) and healthy samples. We demonstrated this by adopting a machine learning model trained with SERS data obtained using cocultured T cells with preclinical models (CAT, CSCAT) of primary (H69AR) and metastatic lung cancer (H1915). Interrogating these distinct signatures with PDT captured the complexity and diversity of the tumor-associated T cell signature across the patient population, exposing the clinical feasibility of immune diagnosis in an independent cohort of patient samples. Thus, our predictive approach using T cells from the patient peripheral blood showed a highly accurate diagnosis with a specificity and sensitivity of 94.1% and 100%, respectively, for primary lung cancer and 97.9% and 94.4% for metastatic lung cancer. Our results prove that the immune-diagnostic signature developed in this study could be used as a clinical technology for cancer diagnosis and determine the course of clinical management with T cells.

CD99 and NKX2.2 positive neuroblastoma diagnosed on cytology: A potential diagnostic pitfall and necessity of pathological evaluation of the primary site.

Immunohistochemistry plays a vital role in the diagnosis of small round cell tumors. CD99 immunonegativity is one of the features, which helps in distinguishing neuroblastoma from other small round cell tumors. NKX2.2 is a specific marker of Ewing sarcoma, which is a differential for poorly differentiated neuroblastoma. Here, we present a case of metastatic neuroblastoma showing immunoreactivity for both CD99 and NKX2.2 on cytology of the metastatic site causing diagnostic dilemma. Biopsy study of the adrenal lesion revealed presence of differentiating cells and neuropil, highlighting the importance of evaluation of the primary site and limitation of cytology.

Molecular Crosstalk between T Cells and Tumor Uncovers GBM-Specific T Cell Signatures in Blood: Noninvasive GBM Diagnosis Using Immunosensors.

Glioblastoma (GBM) is the most common and aggressive stage IV brain cancer with a poor prognosis and survival rate. The blood-brain barrier (BBB) in GBM prevents the entry and exit of biomarkers, limiting its treatment options. Hence, GBM diagnosis is pivotal for timely clinical management. Currently, there exists no clinically validated biomarker for GBM diagnosis. T cells exhibit the potential to escape a leaky BBB in GBM patients. These T cells infiltrating the GBM interact with the heterogeneous population of tumor cells, display a symbiotic interaction resulting in intertwined molecular crosstalk, and display a GBM-associated signature while entering the peripheral circulation. Therefore, we hypothesize that studying these distinct molecular changes is critical to enable T cells to be a diagnostic marker for accurate detection of GBM from patient blood. We demonstrated this by utilizing the phenotypic and immunological landscape changes in T cells associated with glioblastoma tumors. GBM exhibits a high level of heterogeneity with diverse subtypes of cells within the tumor, enabling immune infiltration and different degrees of interactions with the tumor. To accurately detect these subtle molecular differences in T cells, we designed an immunosensor with a high detection sensitivity and repeatability. Hence in this study, we investigated the characteristic behavior of T cells to establish two preclinically validated biomarkers: GBM-associated T cells (GBMAT) and GBM stem cell-associated T cells (GSCAT). A comprehensive investigation was conducted by mimicking the tumor microenvironment in vitro by coculturing T cells with cancer cells and cancer stem cells to study the distinct variation in GBMAT and GSCAT. Preclinical investigation of T cells from GBM patient blood shows similar characteristics to our established biomarkers (GBMAT, GSCAT). Further evaluating the relative attributes of T cells in patient blood and tissue biopsy confirms the infiltrating ability of T cells across the BBB. A pilot validation using a SERS-based machine learning algorithm was accomplished by training the model with GBMAT and GSCAT as diagnostic markers. Using GBMAT as a biomarker, we achieved a sensitivity and specificity of 93.3% and 97.4%, respectively, whereas applying GSCAT yielded a sensitivity and specificity of 100% and 98.7%, respectively. We also validated this diagnostic methodology by using conventional biological assays to study the change in expression levels of T cell surface markers (CD4 and CD8) and cytokine levels in T cells (IL6, IL10, TNFα, INFγ) from GBM patients. This study introduces T cells as GBM-specific immune biomarkers to diagnose GBM using patient liquid biopsy. This preclinical validation study presents a better translatability into clinical reality that will enable rapid and noninvasive glioblastoma detection from patient blood.

Nanoplatform to Investigate Tumor-Initiating Cancer Stem Cells: Breaking the Diagnostic Barrier.

Drug-resistant capacity in a small population of tumor-initiating cancer stem cells (tiCSCs) can be due to aberrant epigenetic changes. However, currently available conventional detection methods are inappropriate and cannot be applied to investigate the scarce population (tiCSCs). In addition, selective inhibitor drugs are shown to reverse epigenetic changes; however, each cancer type is discrete. Hence, it is essential to probe the resultant changes in tiCSCs even after therapy. Therefore, we have developed a multimode nanoplatform to investigate tiCSCs, detect epigenetic changes, and subsequently explore their transformation signals following drug therapy. We performed this by developing a surface-enhanced Raman scattering (SERS)-active nanoplatform integrated with n-dopant using an ultrafast laser ionization technique. The dopant functionalization enhances Raman scattering ability and permits label-free analysis of biomarkers in tiCSCs with the resolution down to the cellular level. Here, we investigated epigenetic biomarkers of tiCSCs in pancreatic and lung cancers. An extended study using inhibitor drugs demonstrates an unexpected increase of tiCSCs from lung cancer; this difference can be attributed to transformation changes in lung tiCSC. Thus, our work brings new insight into the differentiation abilities of CSCs upon epigenetic reversal, emphasizing unique perceptions in cancer treatment.

Doing More with Less: Fine Needle Aspiration Cytology in Pediatric Neoplasms.

BACKGROUND: Cancer is one of the leading causes of death in the pediatric age group following infections. Among the diagnostic modalities, fine needle aspiration cytology (FNAC) is increasingly recognized as it permits rapid diagnosis with low cost and complication. In this study, we emphasize the diagnostic value of FNAC and describe the cytomorphological spectrum of tumors diagnosed on FNAC in pediatric patients with or without the aid of ancillary tests. MATERIALS AND METHODS: This retrospective study included a total of 614 patients under the age of 15 years for whom fine needle aspiration (FNA) was done during a period of 3 years with or without guidance. The cytology smears were reviewed, and the morphological spectrum was analyzed with the ancillary studies. RESULTS: Aspirates from children constituted around 3.5% of the total FNAC performed in our Institute. Of the 614 cases, 336 were male, and 278 were female with age under 15 years. Neoplastic cases constituted around 72%, which included benign (2%) and malignant (98%) tumors. The spectrum include hematolymphoid neoplasms in 39.3%, small round cell tumors (SRCT) in 24.9%, Wilms tumor in 9.2%, germ cell tumors in 4.8%, spindle cell neoplasms in 4.8%, hepatoblastoma in 3.2%, and osteosarcoma in 3% of the cases. The metastatic lesions constituted 8.1% of the cases diagnosed by FNA. CONCLUSION: FNA proves to be a reliable and efficien modality in diagnosing pediatric neoplasms in the hands of a skilled cytopathologist.

Predicting Metastasis from Cues of Metastatic Cancer Stem-like Cells-3D-Ultrasensitive Metasensor at a Single-Cell Level.

Metastasis is the primary reason for treatment failure and cancer-related deaths. Hence forecasting the disease in its primary state can advance the prognosis. However, existing techniques fail to reveal the tumor heterogeneity or its evolutionary cascades; hence they are not feasible to predict the onset of metastatic cancer. The key to metastasis originates from the primary tumor cells, evolving by inheriting multistep sequential cue signals. We have identified this specific population, termed metastatic cancer stem-like cells (MCSCs), to foresee cancer's ability to metastasize. An invasive property renders MCSCs nonadherent, summoning a powerful technique to forecast metastasis. Thus, we have generated an ultrasensitive 3D-metasensor to efficiently capture and investigate MCSCs and magnify the vital premetastatic signals from a single cell. We developed 3D-metasensor by an ultrafast laser ionization technique, consisting of self-assembled three-dimensionally organized nanoprobes incorporated with dopant functionalities. This distinct methodology establishes attachment with nonadherent MCSCs, elevates Raman activity, and enables probing of consequent signals (metabolic, proliferation, and metastatic) specifically altered in MCSCs. Extensive analysis using prediction tools-the area under the curve (AUC) and principal component analysis (PCA)-revealed high sensitivity (100%) and specificity (80%) to differentiate the MCSCs from other populations. Further, investigation reveals that the cue signal level from MCSCs of primary cancer is analogous to MCSCs from higher-level tumors, disclosing the relative dependence to estimate the primary tumor's capacity to metastasize. Multiple spectrum evaluation using the metasensor pinpoint the dynamic cues in MCSCs predict the onset of metastasis; thus, exploring these metastasis hallmarks can enhance prognosis and revolutionize therapy strategies.

Correction to "Green Transfection: Cationic Lipid Nanocarrier System Derivatized from Vegetable Fat, Palmstearin Enhances Nucleic Acid Transfections".

[This corrects the article DOI: 10.1021/acsomega.7b00935.].

Primary Ewing sarcoma of the lung diagnosed on cytology: Case report with review of literature.

Primary Ewing sarcoma (ES) of the lung is anecdotally rare, with few cases reported in literature. This report describes a 46 year-old man who presented with cough and chest pain. CT Thorax revealed a lesion in the right lung. Ultrasound guided fine-needle aspiration of the mass and subsequent cytologic examination exhibited a small round cell morphology. Immunohistochemistry done on the cell block revealed CD 99 and FLI-1 positivity, confirming the diagnosis of ES. FISH supported the diagnosis, showing the EWSR1 rearrangement. Radiologic investigations ruled out a primary tumour elsewhere, confirming the diagnosis of primary pulmonary ES. The patient was started on chemotherapy.

Probing Cancer Metastasis at a Single-Cell Level with a Raman-Functionalized Anionic Probe.

Cancer metastasis is the primary reason for cancer-related deaths, yet there is no technique capable of detecting it due to cancer pathogenesis. Current cancer diagnosis methods evaluate tumor samples as a whole/pooled sample process loses heterogeneous information in the metastasis state. Hence, it is not suitable for metastatic cancer detection. In order to gain complete information on metastasis, it is desirable to develop a nondestructive detection method that can evaluate metastatic cells with sensitivity down to single-cell resolution. Here we demonstrated self-functionalized anionic quantum probes for in vitro metastatic cancer detection at a single-cell concentration. We achieved this by incorporating a nondestructive SERS ability within the generated probes by integrating anionic surface species and NIR plasmon resonance. To the best of our knowledge, this was the first time that metastatic cancer cells were detected through their neoplastic transformations. With reliable diagnostic information at the single-cell sensitivity in an in vitro state, we successfully discriminated against cancer malignancy states.

An anti-oxidant, α-lipoic acid conjugated oleoyl-sn-phosphatidylcholineas a helper lipid in cationic liposomal formulations.

Development of safe non-viral carrier systems for efficient intra-cellular delivery of drugs and genes hold promise in the area of translational research. Liposome based delivery systems have emerged as one of the attractive strategies for efficient delivery of drugs and nucleic acids. To this end, number of investigations was carried on liposomal formulations using lipids for achieving higher efficiency in transfection with lower cytotoxicities. In our efforts to develop safer and efficient liposomal delivery systems, we synthesized a novel anti-oxidant lipid, α-lipoyl, oleyl-sn-phosphatidylcholine (LOPC) and used as a helper lipid in combination with a cationic amphiphile, Di-Stearyl Dihydroxy Ethyl Ammonium Chloride (DSDEAC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) at varying concentrations of LOPC. DNA binding properties of the liposomal formulations (DS, DS LA1, DS LA2 and DS LA3) revealed that increasing the percentage of single aliphatic chain lipid LOPC, did not affect the DNA binding properties. But, transfection profiles of these liposomal formulations in 3 different cell lines (HeLa, HEK 293 and MCF7) showed difference in their efficacies. Results showed that optimal percentage of LOPC i.e. 25% in DSDEAC and DOPC at 1:1 molar ratio (DS LA1) enhanced transfection as compared to DSDEAC:DOPC alone. The endosomal escape studies with NBD labelled lysotracker and Rhodamine labelled liposomal formulations revealed that DS LA1 and DS LA2 facilitated the release of genetic cargo with a better efficiency than their counter parts. Reactive Oxygen Species (ROS), a key modulator of necroptosis were lowered with the treatment of DS LA1 than other liposomal formulations. Here in, we present a novel liposomal formulation using DSDEAC and DOPC at 1:1 molar ratio doped with 25-50% (mole ratio) LOPC as an efficient delivery system for enhanced transfection with quenching of ROS levels compared to formulations without LOPC.

Green Transfection: Cationic Lipid Nanocarrier System Derivatized from Vegetable Fat, Palmstearin Enhances Nucleic Acid Transfections.

Cationic lipid-guided nucleic acid delivery holds great promise in gene therapy and genome-editing applications for treating genetic diseases. However, the major challenge lies in achieving therapeutically relevant efficiencies. Prior findings, including our own, demonstrated that asymmetry in the hydrophobic core of cationic lipids imparted superior transfection efficiencies. To this end, we have developed a lipid nanocarrier system with an asymmetric hydrophobic core (PS-Lips) derived from a mixture of fatty acids of food-grade palmstearin and compared its efficiency with symmetric palmitic acid-based nanocarrier system (P-Lip). PS-Lips exhibited superior transfection efficiencies with both plasmid DNA (pDNA) and mRNA in multiple cultured cells than the control P-Lip. More importantly, PS-Lips exhibited 2-fold superior transfections with linear nucleic acid, green fluorescent protein (GFP) mRNA in hematopoietic cells, when compared with the commercial control lipofectamine RNAiMAX. PS-Lips was also found to be effective in delivering genome-editing tools (CRISPR/Cas9, sgRNA encoded pDNA with a reporter GFP construct) than P-Lip in HEK-293 cells. In the present study, we report that cationic liposomes derivatized from natural food-grade fat palmstearin with a natural hydrophobic core asymmetry are efficient in delivering both linear and circular nucleic acids. In particular, PS-Lips is efficient in delivering mRNA to hematopoietic cells. These findings can be further exploited in the genome-editing approach for treating ß-globinopathies.

Vascular endothelial growth factor expression and angiogenesis in various grades and subtypes of meningioma.

BACKGROUND: Vascular endothelial growth factor (VEGF) expression has been extensively studied in astrocytoma, whereas relatively less literature exists on VEGF expression in meningioma. MATERIALS AND METHODS: Patients operated for meningioma from 2006 to 2011 (n = 46) were included. Tumor was subtyped and graded as per WHO grading. Immunohistochemistry was performed for MIB labeling index, VEGF, and CD 34 staining. The patterns of VEGF expression in various histological subtypes and grades and its correlation with microvascular density were analyzed. RESULTS: This series consisted of 40 Grade I meningioma, 4 Grade II tumors, and 2 Grade III tumors. While 14 (30.4%) tumors showed no staining with VEGF antibody, 32 (69.6%) were positive for VEGF. Sixty five percent of Grade I tumors showed VEGF positivity, while 100% of Grade II and Grade III tumors were VEGF positive (P = 0.157). The mean microvascular density in VEGF-negative tumors was 9.00, while that of VEGF-positive tumors was 17.81(P = 0.013). There was a gradual increase in microvascular density from tumors which are negative for VEGF to tumors which expressed moderate to strong VEGF, the difference being statistically significant (P = 0.009). CONCLUSIONS: VEGF expression correlated with the microvascular density in meningioma irrespective of tumor grade, with a gradual increase in microvascular density in relation to the VEGF score.