7 results on '"Rao, Abhishek S."'
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2. Detection of Epithelial Giant Cells in Nasal Aspirate Cytological Smears Using Deep Learning and Computer Vision Techniques: an Approach for Early Diagnosis of Measles Disease.
- Author
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Rao, Abhishek S., B. H., Karthik Pai, Nayak, Sneha, and Shenoy, Rathika D.
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EPITHELIAL cells , *MULTINUCLEATED giant cells , *DIAGNOSIS , *DEEP learning , *COMPUTER vision , *EARLY diagnosis , *CELL size - Abstract
Measles disease is usually diagnosed through an RT-PCR test, which detects the measles-specific IgM antibody in serum samples. However, this approach is highly expensive and requires a lot of sample preprocessing making it a laborious task. Another approach for measles identification could be a cytological smear test -- which is a low-cost, quick, and accurate method for the early detection of measles disease. Due to the lack of experience in interpreting smears, its usage in clinical practice is currently limited. We have designed a MesoSpot, an automatic segmentation method for the first time that can precisely define the region of giant cells in nasal aspirate cytological smears and assist medical experts in the speedy diagnosis of measles disease. MesoSpot was developed by training 500 cytological smear images collected from various web sources. We have used 50 cytological smear images to validate the model and evaluated its performance using various metrics. The main purpose of the proposed study is to detect multinucleated giant cells and estimate their size and number, which aids in assessing the disease's severity. To annotate the giant cells in each cytological smear, we have applied a semantic segmentation approach using APEER software. UNET architecture was used to train the model, and image processing techniques were used to spot giant cells and predict their size. Various performance metrics were employed to assess the model's efficiency. The model's output was also compared to the ground truth image and statistically confirmed. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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3. Swift Automated System for Distinguishing Blue-White Colonies Post Bacterial Transformation on Agar Plates Using Computer Vision Techniques.
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Rao, Abhishek S., Goveas, Louella Concepta, and Nayak, Sneha
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BACTERIAL transformation , *COMPUTER vision , *BACTERIAL colonies , *AGAR plates , *MICROCYSTIS , *CELL size - Abstract
The manual selection of transformed bacterial colonies from non-transformed ones grown on agar plate post-blue-white screening -- despite chromogenic difference -- is cumbersome, owing to their small size and large cell number. The present study offers a lucrative solution to this problem by the design of an automated system that is not only fast and less laborious but also low priced and user-friendly. The image masking technique was used to distinguish plated transformed colonies. This method uses computer vision techniques to detect the number of blue and white colonies post bacterial transformation on agar plates and calculate the transformation efficiency. To assess the proposed model with the manual counting method, we have validated the model by comparing the manual counting of colonies with the automated system count. Hence, a model was developed that would be an added advantage to biotechnologists as it would minimize the time required for counting and help in productive research. [ABSTRACT FROM AUTHOR]
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- 2021
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4. Phenotypic and functional analysis of malignant mesothelioma tumor-infiltrating lymphocytes.
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Klampatsa, Astero, O'Brien, Shaun M., Thompson, Jeffrey C., Rao, Abhishek S., Stadanlick, Jason E., Martinez, Marina C., Liousia, Maria, Cantu, Edward, Cengel, Keith, Moon, Edmund K., Singhal, Sunil, Eruslanov, Evgeniy B., and Albelda, Steven M.
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FUNCTIONAL analysis ,LYMPHOCYTES ,PROGRAMMED cell death 1 receptors ,CELL suspensions ,TRANSCRIPTION factors - Abstract
Given the growing interest and promising preliminary results of immunotherapy in malignant pleural mesothelioma (MPM), it has become important to more fully understand the immune landscape in this tumor. This may be especially relevant in deciding who might benefit most from checkpoint blockade or agonist antibody therapy. Since the phenotype of tumor infiltrating lymphocytes (TILs) in MPM has not been fully described and their function has not been carefully assessed, we collected fresh tumor and blood from 22 patients undergoing surgical resection and analysed single cell suspensions by flow cytometry. The functionality of TILs was assessed by measurement of cytokine expression (IFN-γ) following overnight stimulation ex vivo. Results showed low numbers of CD8+ TILs whose function was either moderately or severely suppressed. The degree of TIL hypofunction did not correlate with the presence of co-existing macrophages or neutrophils, nor with expression of the inhibitory receptors PD-1, CD39 and CTLA-4. Hypofunction was associated with higher numbers of CD4 regulatory T cells (Tregs) and with expression of the inhibitory receptor TIGIT. On the other hand, presence of tissue-resident memory (Trm) cells and expression of TIM-3 on CD8+ cells were positively associated with cytokine production. However, Trm function was partially suppressed when the transcription factor Eomesodermin (Eomes) was co-expressed. Understanding the function of TILs in malignant mesothelioma may have clinical implications for immunotherapy, especially in choosing the best immunotherapy targets. Our data suggests that Treg cell blocking agents or TIGIT inhibitor antibodies might be especially valuable in these patients. [ABSTRACT FROM AUTHOR]
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- 2019
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5. Human tumor-associated monocytes/macrophages and their regulation of T cell responses in early-stage lung cancer.
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Singhal, Sunil, Stadanlick, Jason, Annunziata, Michael J., Rao, Abhishek S., Bhojnagarwala, Pratik S., O'Brien, Shaun, Moon, Edmund K., Cantu, Edward, Danet-Desnoyers, Gwenn, Ra, Hyun-Jeong, Litzky, Leslie, Akimova, Tatiana, Beier, Ulf H., Hancock, Wayne W., Albelda, Steven M., and Eruslanov, Evgeniy B.
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MONOCYTES ,MACROPHAGES ,T cells ,IMMUNOSUPPRESSIVE agents ,LUNG cancer ,LUNG diseases - Abstract
PD-L1
+ tumor cells, but not macrophages, are likely responsible for inhibiting tumor-specific T cells in early-stage human lung tumors. In defense of tumor-associated macrophages: Although preclinical models using implanted tumors in mice are valuable, they cannot completely recapitulate the early stages of natural tumor development in humans. To better understand how monocytes and macrophages influence developing tumor immunity, Singhal et al. studied samples from patients with early-stage lung cancer. They phenotyped monocytes and macrophages in tumors and adjacent tissue, as well as samples from control subjects without cancer. The authors found that, although tumor-associated macrophages expressed PD-L1, these cells did not generally suppress T cell responses. Their results suggest that tumor-associated macrophages should not be assumed to be protumorigenic, especially during early stages of cancer. Data from mouse tumor models suggest that tumor-associated monocyte/macrophage lineage cells (MMLCs) dampen antitumor immune responses. However, given the fundamental differences between mice and humans in tumor evolution, genetic heterogeneity, and immunity, the function of MMLCs might be different in human tumors, especially during early stages of disease. Here, we studied MMLCs in early-stage human lung tumors and found that they consist of a mixture of classical tissue monocytes and tumor-associated macrophages (TAMs). The TAMs coexpressed M1/M2 markers, as well as T cell coinhibitory and costimulatory receptors. Functionally, TAMs did not primarily suppress tumor-specific effector T cell responses, whereas tumor monocytes tended to be more T cell inhibitory. TAMs expressing relevant MHC class I/tumor peptide complexes were able to activate cognate effector T cells. Mechanistically, programmed death-ligand 1 (PD-L1) expressed on bystander TAMs, as opposed to PD-L1 expressed on tumor cells, did not inhibit interactions between tumor-specific T cells and tumor targets. TAM-derived PD-L1 exerted a regulatory role only during the interaction of TAMs presenting relevant peptides with cognate effector T cells and thus may limit excessive activation of T cells and protect TAMs from killing by these T cells. These results suggest that the function of TAMs as primarily immunosuppressive cells might not fully apply to early-stage human lung cancer and might explain why some patients with strong PD-L1 positivity fail to respond to PD-L1 therapy. [ABSTRACT FROM AUTHOR]- Published
- 2019
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6. Origin and Role of a Subset of Tumor-Associated Neutrophils with Antigen-Presenting Cell Features in Early-Stage Human Lung Cancer.
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Singhal, Sunil, Bhojnagarwala, Pratik S., O'Brien, Shaun, Moon, Edmund K., Garfall, Alfred L., Rao, Abhishek S., Quatromoni, Jon G., Stephen, Tom Li, Litzky, Leslie, Deshpande, Charuhas, Feldman, Michael D., Hancock, Wayne W., Conejo-Garcia, Jose R., Albelda, Steven M., and Eruslanov, Evgeniy B.
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TUMORS , *NEUTROPHILS , *ANTIGEN presenting cells , *LUNG cancer , *PROGENITOR cells - Abstract
Summary Based on studies in mouse tumor models, granulocytes appear to play a tumor-promoting role. However, there are limited data about the phenotype and function of tumor-associated neutrophils (TANs) in humans. Here, we identify a subset of TANs that exhibited characteristics of both neutrophils and antigen-presenting cells (APCs) in early-stage human lung cancer. These APC-like “hybrid neutrophils,” which originate from CD11b + CD15 hi CD10 − CD16 low immature progenitors, are able to cross-present antigens, as well as trigger and augment anti-tumor T cell responses. Interferon-γ and granulocyte-macrophage colony-stimulating factor are requisite factors in the tumor that, working through the Ikaros transcription factor, synergistically exert their APC-promoting effects on the progenitors. Overall, these data demonstrate the existence of a specialized TAN subset with anti-tumor capabilities in human cancer. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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7. Human Tumor-Associated Macrophages and Neutrophils Regulate Antitumor Antibody Efficacy through Lethal and Sublethal Trogocytosis.
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Singhal S, Rao AS, Stadanlick J, Bruns K, Sullivan NT, Bermudez A, Honig-Frand A, Krouse R, Arambepola S, Guo E, Moon EK, Georgiou G, Valerius T, Albelda SM, and Eruslanov EB
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- Humans, Tumor-Associated Macrophages metabolism, Trogocytosis, Antibody-Dependent Cell Cytotoxicity, Phagocytosis, Receptors, Fc, Antigens, Neoplasm, Neutrophils metabolism, Neoplasms pathology
- Abstract
The clinical benefits of tumor-targeting antibodies (tAb) are modest in solid human tumors. The efficacy of many tAbs is dependent on Fc receptor (FcR)-expressing leukocytes that bind Fc fragments of tAb. Tumor-associated macrophages (TAM) and neutrophils (TAN) represent the majority of FcR+ effectors in solid tumors. A better understanding of the mechanisms by which TAMs and TANs regulate tAb response could help improve the efficacy of cancer treatments. Here, we found that myeloid effectors interacting with tAb-opsonized lung cancer cells used antibody-dependent trogocytosis (ADT) but not antibody-dependent phagocytosis. During this process, myeloid cells "nibbled off" tumor cell fragments containing tAb/targeted antigen (tAg) complexes. ADT was only tumoricidal when the tumor cells expressed high levels of tAg and the effectors were present at high effector-to-tumor ratios. If either of these conditions were not met, which is typical for solid tumors, ADT was sublethal. Sublethal ADT, mainly mediated by CD32hiCD64hi TAM, led to two outcomes: (i) removal of surface tAg/tAb complexes from the tumor that facilitated tumor cell escape from the tumoricidal effects of tAb; and (ii) acquisition of bystander tAgs by TAM with subsequent cross-presentation and stimulation of tumor-specific T-cell responses. CD89hiCD32loCD64lo peripheral blood neutrophils (PBN) and TAN stimulated tumor cell growth in the presence of the IgG1 anti-EGFR Ab cetuximab; however, IgA anti-EGFR Abs triggered the tumoricidal activity of PBN and negated the stimulatory effect of TAN. Overall, this study provides insights into the mechanisms by which myeloid effectors mediate tumor cell killing or resistance during tAb therapy., Significance: The elucidation of the conditions and mechanisms by which human FcR+ myeloid effectors mediate cancer cell resistance and killing during antibody treatment could help develop improved strategies for treating solid tumors., (©2024 The Authors; Published by the American Association for Cancer Research.)
- Published
- 2024
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