Distribution Pattern Of Cells Research Articles

Tension force causes cell cycle arrest at G2/M phase in osteocyte-like cell line MLO-Y4

Bone remodelling is the process of bone resorption and formation, necessary to maintain bone structure or for adaptation to new conditions. Mechanical loadings, such as exercise, weight bearing and orthodontic force, play important roles in bone remodelling. During the remodelling process, osteocytes play crucial roles as mechanosensors to regulate osteoblasts and osteoclasts. However, the precise molecular mechanisms by which the mechanical stimuli affect the function of osteocytes remain unclear. In the present study, we analysed viability, cell cycle distribution and gene expression pattern of murine osteocyte-like MLO-Y4 cells exposed to tension force (TF). Cells were subjected to TF with 18% elongation at 6 cycles/min for 24 h using Flexcer Strain Unit (FX-3000). We found that TF stimulation induced cell cycle arrest at G2/M phase but not cell death in MLO-Y4 cells. Differentially expressed genes (DEGs) between TF-stimulated and unstimulated cells were identified by microarray analysis, and a marked increase in glutathione-S-transferase α (GSTA) family gene expression was observed in TF-stimulated cells. Enrichment analysis for the DEGs revealed that Gene Ontology (GO) terms and Kyoto Encyclopedia Genes and Genomes (KEGG) pathways related to the stress response were significantly enriched among the upregulated genes following TF. Consistent with these results, the production of reactive oxygen species (ROS) was elevated in TF-stimulated cells. Activation of the tumour suppressor p53, and upregulation of its downstream target GADD45A, were also observed in the stimulated cells. As GADD45A has been implicated in the promotion of G2/M cell cycle arrest, these observations may suggest that TF stress leads to G2/M arrest at least in part in a p53-dependent manner.

Multi-omics analyses reveal the difference of stone cell distribution in pear fruit.

Stone cells are the brachysclereid cells in pear (Pyrus) fruit, consisting almost entirely of lignified secondary cell walls. They are distributed mainly near the fruit core and spread radially in the whole fruit. However, the development of stone cells has not been comprehensively characterized. The regulation of stone cell formation on transcriptomic, proteomic and metabolomic levels remains largely unknown. In this study, we performed phenomic analysis on the stone cells and their associated vascular bundles distributed near the fruit cores. Transcriptomic, proteomic and metabolomic analyses revealed significantly positive regulation of biological processes, which contributed to the lignification and lignin deposition in stone cells near the fruit core, including sucrose metabolism, phenylalanine, tyrosine, tryptophan, and phenylalanine biosynthesis. We then found many metabolites generated from the phenylpropanoid pathway, contributing to the cell wall formation of stone cells near the fruit core. Furthermore, we identified a key transcription factor, PbbZIP48, which was significantly highly expressed near the fruit core and was then proven to regulate the lignin biosynthesis in stone cells. In conclusion, we provide new insights into understanding the mechanism of lignified stone cell formation near the pear fruit core at multi-omics levels.

Histamine and gamma-aminobutyric acid in the nervous system of Pygospio elegans (Annelida: Spionidae): structure and recovery during reparative regeneration

BackgroundIn recent two decades, studies of the annelid nervous systems were revolutionized by modern cell labeling techniques and state-of-the-art microscopy techniques. However, there are still huge gaps in our knowledge on the organization and functioning of their nervous system. Most of the recent studies have focused on the distribution of serotonin and FMRFamide, while the data about many other basic neurotransmitters such as histamine (HA) and gamma-aminobutyric acid (GABA) are scarce.ResultsUsing immunohistochemistry and confocal microscopy we studied the distribution of histamine and gamma-aminobutyric acid in the nervous system of a spionid annelid Pygospio elegans and traced their redevelopment during reparative regeneration. Both neurotransmitters show specific patterns in central and peripheral nervous systems. HA-positive cells are concentrated mostly in the brain, while GABA-positive cell somata contribute equally to brain and segmental ganglia. Some immunoreactive elements were found in peripheral nerves. Both substances were revealed in high numbers in bipolar sensory cells in the palps. The first signs of regenerating HAergic and GABAergic systems were detected only by 3 days after the amputation. Further redevelopment of GABAergic system proceeds faster than that of HAergic one.ConclusionsComparisons with other annelids and mollusks examined in this respect revealed a number of general similarities in distribution patterns of HAergic and GABAergic cells in different species. Overall, the differences in the full redevelopment of various neurotransmitters correlate with neuronal development during embryogenesis. Our results highlight the importance of investigating the distribution of different neurotransmitters in comparative morphological and developmental studies.

Barcode clonal tracking of tissue-resident immune cells in rhesus macaque highlights distinct clonal distribution pattern of tissue NK cells.

Tissue resident (TR) immune cells play important roles in facilitating tissue homeostasis, coordinating immune responses against infections and tumors, and maintaining immunological memory. While studies have shown these cells are distinct phenotypically and functionally from cells found in the peripheral blood (PB), the clonal relationship between these populations across tissues has not been comprehensively studied in primates or humans. We utilized autologous transplantation of rhesus macaque hematopoietic stem and progenitor cells containing high diversity barcodes to track the clonal distribution of T, B, myeloid and natural killer (NK) cell populations across tissues, including liver, spleen, lung, and gastrointestinal (GI) tract, in comparison with PB longitudinally post-transplantation, in particular we focused on NK cells which do not contain endogenous clonal markers and have not been previously studied in this context. T cells demonstrated tissue-specific clonal expansions as expected, both overlapping and distinct from blood T cells. In contrast, B and myeloid cells showed a much more homogeneous clonal pattern across various tissues and the blood. The clonal distribution of TR NK was more heterogenous between individual animals. In some animals, as we have previously reported, we observed large PB clonal expansions in mature CD56-CD16+ NK cells. Notably, we found a separate set of highly expanded PB clones in CD16-CD56- (DN) NK subset that were also contributing to TR NK cells in all tissues examined, both in TR CD56-CD16+ and DN populations but absent in CD56+16- TR NK across all tissues analyzed. Additionally, we observed sets of TR NK clones specific to individual tissues such as lung or GI tract and sets of TR NK clones shared across liver and spleen, distinct from other tissues. Combined with prior functional data that suggests NK memory is restricted to liver or other TR NK cells, these clonally expanded TR NK cells may be of interest for future investigation into NK cell tissue immunological memory, with implications for development of NK based immunotherapies and an understanding of NK memory.

Toxic Dinophysis acuminata in southern Chile: A comparative approach based on its first local in vitro culture and offshore-estuarine bloom dynamics

In Chile, Dinophysis acuminata is a conspicuous mixotrophic dinoflagellate that inhabits the Offshore Pacific Coast (OPC) and the Patagonian fjords (PF). In this study, we investigated D. acuminata phylogeny, morphology, and the effect of environmental factors on cell growth and toxin production of isolates from the OPC and the PF. A comparative approach was performed between in vitro cultures, maintained for the first time in Chile, and phytoplankton field monitoring. Phylogenetic analyses revealed that Chilean D. acuminata sequences clustered in the “Dinophysis acuminata complex” with cell morphology consistent with the “Type A” morphotype. Different reaction norms among the strains from the different oceanic conditions were observed, with the highest growth rate and maximum cell density of the strain isolated from the PF, cultured at 15 °C of temperature and 95 μmol m−2 s−1 light intensity. Pectenotoxin 2 (PTX2) was the only toxin detected in all strains isolated from both OPC and PF. Monitoring data (from 2013 to 2021) of D. acuminata and its prey Mesodinium rubrum were also analyzed to determine the effect of temperature, salinity, depth, and geographic distribution on species abundance and presence probability (PP), along with potential links with climatic patterns such as the Southern Annular Mode (SAM) and El Niño Southern Oscillation. Cell distribution patterns of D. acuminata and M. rubrum were highly related and influenced by low salinity and high temperature in the estuarine zone (PF). Meanwhile, a negative relationship between SAM and D. acuminata abundance was observed. Coupling, the first local D. acuminata culture and field data, allows a better understanding of the spatial–temporal and environmental effects on D. acuminata distribution and growth.

Foliar morphology and epidermal ultrastructure of four Selaginella species (Selaginellaceae – Lycophyta)

ABSTRACT The leaf morphology of Selaginella species from subtropical South America show differences in structure and ultrastructure that are useful for species identification. We studied the following epidermal traits: micromorphology of stomatal types, distribution patterns of silica cells, silica cells and silica bodies of Selaginella convoluta, S. microphylla, S. sellowii and S. sulcata, using light and scanning electron microscopy. Five stomatal types were observed, anomocytic, actinocytic, anisocytic, staurocytic and tetracytic and two additional variants, anomodiacytic and anomoparacytic. Five silica cell distribution patterns were identified, laminar, medial, medial-basiscopic, subcostal, and submarginal. Pyramidal silica bodies and conical bodies with concave base are described for the first time. In the studied species, silica bodies are pyramidal in S. convoluta and globular, conical and pyramidal in S. microphylla, whereas in S. sellowii and S. sulcata they are conical, with concave base of different sizes. Illustrations of the epidermal features, LM photographs, SEM micrographs and comparative tables are presented.

Clinical usefulness of a novel classification of T cell distribution patterns in the tumor microenvironment of follicular lymphoma to detect transformation.

The clinical course of follicular lymphoma (FL) is thought to be influenced by the infiltrating immune cells in the tumor microenvironment. Focusing on the distribution patterns of T cells may be a promising approach to estimate the prognosis of FL, especially histological transformation. This study was a retrospectively cohort study in the relationship between the pathological distribution pattern of T cells in the tumor microenvironment and clinical course of FL. One hundred twenty-eight patients with FL initially diagnosed at the University of Tokyo Hospital from January 2008 to January 2017 were evaluated. We classified each patient's specimen at initial diagnosis by the distribution pattern of tumor infiltrating CD3-positive cells, intra-follicle focal (IFF), intra-follicle diffuse (IFD), extra-follicle marginal (EFM), and extra-follicle diffuse (EFD). We analyzed the distribution pattern's correlation with other prognostic factors including overall survival (OS), progression free survival (PFS), and transformation. Among 128 cases, 81 had evaluable pathological specimen. Based on our criteria, in the intra-follicle,17 cases (21%) were classified as IFF. Sixty-four cases (79%) were classified as IFD. In the extra follicle, 25 cases (31%) were classified as EFM. Fifty-six cases (69%) were classified as EFD. There was significant difference in risk of transformation between the EFM and EFD around extra-follicle area in the adjusted model (p < 0.05). Also, cases with IFF and EFM had significantly higher risk of transformation compared to cases with other T cell distribution patterns (p < 0.01). We proposed a new classification of CD3-positive T cell distribution patterns around the follicle lesions in FL and demonstrated its clinical significance.

Post-infantile Giant Cell Hepatitis: A Literature Review and Meta-analysis.

Post-infantile giant cell hepatitis (PIGCH) is a rare disease entity in adults with a multifactorial etiology and widely variable clinical courses and outcomes. The factors associated with the worse outcomes of this disease entity are still unclear. We identified 68 PIGCH patients by searching PubMed and performed meta-analysis. Among the 68 patients, 32% of the cases were associated with autoimmune disorders, followed by 21% associated with viral infections, 10% with medication, and 7% with malignancy. Twenty-four percent of the patients had more than one etiological factor, and 6% had other uncommon etiologies or an etiology that could not be identified. At the time of this report, 17 patients had died of the disease (poor outcome), and 51 patients remained alive with the disease (good outcome). Compared to the patients with a good outcome, the patients with a poor outcome were characterized by older age, lower levels of platelets and albumin, higher level of total bilirubin, and a diffuse distribution pattern of giant cells in the liver. There were no differences in gender distribution, aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, etiological distribution, or other histological features, including interface hepatitis, necrosis, lobular inflammation, portal inflammation, cholestasis, or fibrosis. Further studies would be needed to better understand the disease mechanisms and unmask any additional etiological factors and targeted therapies.

Identification of the Novel G250R Variant Indicates a Role for Thrombomodulin in Modulating the Risk for Venous Thromboembolism

Thrombomodulin displays anticoagulant properties through thrombin binding and protein C activation. In close relatives who had a history of recurrent VTE, a missense variant (G250R) in the EGF-1 domain was identified. In an asymptomatic 250R carrier, higher thrombin generation plasma values were found. Evaluation of cells transfected with the wild-type or the mutant construct using imaging technologies and flow cytometry provided evidences that the 250R variant significantly decreased protein localization on cell membrane. The different pattern of cell distribution suggested that the 250R variant is not able to sufficiently target the plasma membrane. Present findings add information to characterize protein structure-function relationships, further suggesting that misfolding of the protein modulates the expression of mature protein. Functional analyses of the 250R variant increase our understanding of the role of Thrombomodulin in coagulation and support the hypothesis that gene variants reducing protein functionality cause deficiency and behave as a thrombophilic risk factor.

Characterization of Organellar-Specific ABA Responses during Environmental Stresses in Tobacco Cells and Arabidopsis Plants.

Abscisic acid (ABA) is a critical phytohormone involved in multifaceted processes in plant metabolism and growth under both stressed and nonstressed conditions. Its accumulation in various tissues and cells has long been established as a biomarker for plant stress responses. To date, a comprehensive understanding of ABA distribution and dynamics at subcellular resolution in response to environmental cues is still lacking. Here, we modified the previously developed ABA sensor ABAleon2.1_Tao3 (Tao3) and targeted it to different organelles including the endoplasmic reticulum (ER), chloroplast/plastid, and nucleus through the addition of corresponding signal peptides. Together with the cytosolic Tao3, we show distinct ABA distribution patterns in different tobacco cells with the chloroplast showing a lower level of ABA in both cell types. In a tobacco mesophyll cell, organellar ABA displayed specific alterations depending on osmotic stimulus, with ABA levels being generally enhanced under a lower and higher concentration of salt and mannitol treatment, respectively. In Arabidopsis roots, cells from both the meristem and elongation zone accumulated ABA considerably in the cytoplasm upon mannitol treatment, while the plastid and nuclear ABA was generally reduced dependent upon specific cell types. In Arabidopsis leaf tissue, subcellular ABA seemed to be less responsive when stressed, with notable increases of ER ABA in epidermal cells and a reduction of nuclear ABA in guard cells. Together, our results present a detailed characterization of stimulus-dependent cell type-specific organellar ABA responses in tobacco and Arabidopsis plants, supporting a highly coordinated regulatory network for mediating subcellular ABA homeostasis during plant adaptation processes.

Distribution pattern of floor plate neurons in zebrafish.

The floor plate (FP) is a critical signaling center for the development of neural tube and body axis, and is localized at the ventral midline of the neural tube. Multiple types of neurons are present in the floor plate, while the distribution pattern of these neuronal cells remains unclear. By using transgenic zebrafish lines that specifically label different neuronal cells, we investigated the distribution pattern of these neurons in the floor plate region. Our results showed that foxj1a, sox2, clusterin and gfap genes were expressed in the medial floor plate (MFP), consisting of a single row of cells. The cerebrospinal fluid-contacting neurons (CSF-cNs), also named as Kolmer-Agduhr interneurons (KA' and KA" neurons), were located on the lateral sides of MFP. The foxj1a and pkd2l1 genes were expressed in the KA" neurons, which were located to the ventral terminal gap of wedge-shaped MFP cells. The neighboring KA" neurons were separated by neurons expressing Gfap, Olig2 or Sox2. In contrast, the KA' neurons were positive for Foxj1a +/Pkd2l1+/Olig2+, and were localized to the dorsal side of KA" neurons. Similarly, the Sox2 or Olig2 expressing neurons were intermingled with KA' neurons along the anterior-posterior axis in these regions. Further pharmaceutical treatment demonstrated that interference of Notch signaling resulted in the abnormal distribution of floor plate neurons together with strong dorsal body curvature at 3 days post fertilization in the zebrafish larvae. Our data showed the gene expression patterns and relative positions of the floor plate neurons; and suggested a potential role of Notch signaling during floor plate development.

Abstract 1710: Spatial transcriptomics and multiplexed imaging to explore tumor heterogeneity and immune complexity

Abstract Spatial transcriptomics (ST) and CO-Detection by indEXing (CODEX) are becoming the leading transformative approaches for studying spatially resolved tumor heterogeneity and immune complexity. To explore the application of these two cutting-edge technologies and to further investigate the complexities of tumor microenvironment, we applied 10x Visium Gene Expression platform and Akoya CODEX platform on surgical resections from pancreatic ductal adenocarcinoma (PDAC), breast cancer (BRCA), and clear cell renal carcinoma (ccRCC) patients. A total of 30+ protein markers were screened, verified, and conjugated as CODEX antibodies for the 3 types of cancers. Tumor microenvironment was vividly revealed by a set of cancer markers, immune markers, and stroma cells markers on a single ST slide and a single CODEX image. Notably, the distinct immune microenvironment was observed between tumor nests in the neighborhood in terms of cell types, number of cells, and cell distribution patterns. This work highlights the importance of direct visualization for tumor heterogeneity. It also provides a significant insight for understanding tumor complexity and a promising opportunity to benefit personalized medicine. Citation Format: Siqi Chen, Andrew Shinkle, Yanyan Zhao, Chia-Kuei Mo, Andrew Houston, Preet Lal, John M. Herndon, Ryan C. Fields, William Gillanders, Feng Chen, Li Ding. Spatial transcriptomics and multiplexed imaging to explore tumor heterogeneity and immune complexity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1710.

Morphometric study of the vestibuloauditory organ of the African clawed frog, Xenopus laevis.

Independent auditory end-organs appear first in amphibians in vertebrate phylogeny. In amphibians, sound detection is carried out by the amphibian papilla, basilar papilla and macula saccule. Amphibians inhabit distinct habitats and exhibit specific behaviours and sound frequency responses, so the amphibian vestibuloauditory system is an excellent model for considering the relationships between behaviour and physiological/anatomical vestibuloauditory properties. The African clawed frog, Xenopus laevis, lives in shallow water throughout its life and is thought to use sound in a higher frequency range compared with terrestrial anurans. In this study, the size of each vestibuloauditory end-organ and the distribution of ganglion cells in the vestibuloauditory ganglion were examined using haematoxylin and eosin staining and lectin histochemistry in Xenopus laevis. This study revealed that the size ratios among end-organs in Xenopus are similar to those in terrestrial anurans. Large and small cells were observed in the ganglion, but their distribution patterns are different from those in general terrestrial anurans. Lycopersicon esculentum lectin stained a large number of ganglion cells. Lectin-stained cells were found throughout the whole ganglion, but were especially abundant in the caudal part. These results suggested a unique distribution pattern of the vestibuloauditory ganglion cells in Xenopus.

INSP-12. Molecular architecture of the human brain and brain cancer

Abstract Single-cell gene expression analysis has revolutionized biomedical research, leading to comprehensive atlases of human tissues, insights into development and homeostasis and allowed highly multiplexed perturbations with cell type specificity. Here, I present our comprehensive single-cell atlas of the human nervous system, both in development and in the adult. We describe unexpected regional diversity among human glial cell types, with implications for regions-specific glial brain disease. However, in single-cell analysis, spatial context is lost, complicating the analysis of cell-cell interactions, spatial gradients and patterns of cell distribution. Spatial methods of gene expression analysis have therefore increased in importance, and are currently a very active are of research and development. We have developed a scalable and highly multiplexed method of spatial gene expression analysis based on electrophoretic capture of mRNA. I showcase its throughput and robustness by presenting complete spatial atlases of the adult mouse brain, as well as the whole developing human head at 7 weeks post-conception. Furthermore, I will discuss our ongoing work to apply these methods to human glioblastoma in order to reveal the spatial architecture of this devastating tumor.

Sex-specific characteristics of cells expressing the cannabinoid 1 receptor in the dorsal horn of the lumbar spinal cord.

It is becoming increasingly clear that robust sex differences exist in the processing of acute and chronic pain in both rodents and humans. However, the underlying mechanism has not been well characterized. The dorsal horn of the lumbar spinal cord is the fundamental building block of ascending and descending pain pathways. It has been shown that numerous neurotransmitter and neuromodulator systems in the spinal cord, including the endocannabinoid system and its main receptor, the cannabinoid 1 receptor (CB1 R), play vital roles in processing nociceptive information. Our previous findings have shown that CB1 R mRNA is widely expressed in the brain in sex-dependent patterns. However, the sex-, lamina-, and cell-type-specific characteristics of CB1 R expression in the spinal cord have not been fully described. In this study, the CB1 R-iCre-EGFP mouse strain was generated to label and identify CB1 R-positive (CB1 RGFP ) cells. We reported no sex difference in CB1 R expression in the lumbar dorsal horn of the spinal cord, but a dynamic distribution within superficial laminae II and III in female mice between estrus and nonestrus phases. Furthermore, the cell-type-specific CB1 R expression pattern in the dorsal horn was similar in both sexes. Over 50% of CB1 RGFP cells were GABAergic neurons, and approximately 25% were glycinergic and 20-30% were glutamatergic neurons. The CB1 R-expressing cells also represented a subset of spinal projection neurons. Overall, our work indicates a highly consistent distribution pattern of CB1 RGFP cells in the dorsal horn of lumbar spinal cord in males and females.

Immunolocalization of endomucin-reactive blood vessels and α-smooth muscle actin-positive cells in murine nasal conchae

ObjectivesRecently, the biological functions of endomucin-positive blood vessels and closely associated αSMA-positive cells in long bones have been highlighted. The surrounding tissues of the flat bones, such as nasal bones covered with mucosa and lamina propria, are different from those of the long bones, indicating the different distributions of endomucin-positive blood vessels and αSMA-reactive cells in nasal bones. This study demonstrates the immunolocalization of endomucin-reactive blood vessels and αSMA-positive cells in the nasal conchae of 3- and 7-week-old mice. MethodsThe nasal conchae of 3-week-old and 7-week-old male C57BL/6J mice were used for immunoreaction of endomucin, CD34, PDGFbb, TRAP, and c-kit. ResultsWhile we identified abundant endomucin-reactive blood vessels in the lamina propria neighboring the bone, not all were positive for endomucin. More CD34-reactive cells and small blood vessels were observed in the nasal conchae of 3-week-old mice than in those of 7-week-old mice. Some αSMA-positive cells in the nasal conchae surrounded the blood vessels, indicating vascular smooth muscle cells, while other αSMA-immunopositive fibroblastic cells were detected throughout the lamina propria. αSMA-positive cells did not co-localize with c-kit-immunoreactivity, thereby indicating that the αSMA-positive cells may be myofibroblasts rather than undifferentiated mesenchymal cells. ConclusionsUnlike long bones, nasal conchae contain endomucin-positive as well as endomucin-negative blood vessels and exhibit numerous αSMA-positive fibroblastic cells throughout the lamina propria neighboring the bone. Apparently, the distribution patterns of endomucin-positive blood vessels and αSMA-positive cells in nasal conchae are different from those in long bones.

Comprehensive Assessment of Selected Immune Cell Subpopulations Changes in Chemotherapy-Naïve Germ Cell Tumor Patients.

The pattern of immune cell distribution in testicular germ cell tumors (GCT) significantly differs from the immune environment in normal testicular tissues. The present study aimed to evaluate the role of different leukocyte subpopulation in GCTs. A cohort of 84 chemotherapy-naïve GCT patients was analyzed. Immunophenotyping of peripheral blood leukocyte subpopulations was carried out by flow cytometry. In addition, the data assessing the immunophenotypes and the baseline clinicopathological characteristics of the included subjects were statistically evaluated. Their prognostic value for the assessment of progression-free survival (PFS) and overall survival (OS) was estimated. The percentage of different innate/adaptive immune cell subpopulations was significantly associated with poor risk-related clinical features, including the number of metastatic sites, presence of retroperitoneal, mediastinal, lung, brain and non-pulmonary visceral metastases as well as with the S-stage and International Germ Cell Consensus Classification Group (IGCCCG) risk groups. In univariate analysis, the percentages of neutrophils, eosinophils, dendritic cells type 2, lymphocytes and T cytotoxic cells were significantly associated with PFS, while the neutrophil, non-classical monocyte and lymphocyte percentage were associated with OS. However, all these outcome correlations were not independent of IGCCCG in multivariate analysis. The data indicated a link among different innate/adaptive peripheral immune cell subpopulations in GCT patients. In addition, the association between these subpopulations and tumor characteristics was also investigated. The findings of the present study may contribute to a deeper understanding of the interactions between cancer and innate/adaptive immune response in GCT patients.

157 Supramarginal Resection Impact on Overall Survival for IDH-Wildtype Glioblastoma According to Their Cell Density Distribution and Infiltration Profile: A Mathematical Model

INTRODUCTION: Supramarginal resection of the T2-FLAIR beyond the T1 contrast enhancement has been suggested to further improve overall survival in glioblastoma (GBM) patients, with a potentially enhanced role in IDH-wildtype. GBMs present with high variability in tumor cell density, distribution, and infiltration patterns evidenced in imaging and histological analysis. Preoperative estimation of GBM grow kinetics with advanced mathematical models can provide further insight for surgical planning. METHODS: Volumetric measurements from T1 contrast-enhancing (CE) and T2-FLAIR regions of interest from retrospective pre and postoperative MRIs of patients that underwent a gross total resection of the CE were obtained. Tumor invasiveness profile (proliferation/diffusion (ρ/D) ratio) was calculated using the following formula: where VFL and VT1 are the preoperative FLAIR and contrast-enhancing volumes, respectively. Patients were then split into nodular (ND), moderately diffuse (MD), and highly diffuse (HD) subgroups based on their tumor invasiveness profile. SMR extension and survival analyses were performed among groups. RESULTS: 101 patients met the criteria for this study. Tumors were classified as ND (n = 34), MD (n = 34), and HD (n = 33) tumors. On multivariate analysis, increasing SMR had a significant positive correlation with OS only in MD and HD tumors (HR 0.98; 95% CI, 0.98–0.99; p = 0.05, and HR 0.98; 95% CI, 0.97–0.99; p = 0.02), respectively. On threshold analysis, OS benefit was seen with SMR from 10% to 29%, 10%-59%, and 30%-90%, for ND, MD, and HD, respectively. CONCLUSION: SMR impact on overall survival in this cohort of IDH-wildtype is significantly influenced by the degree of tumor invasiveness. The results of this study showed increased overall survival with increasing SMR for moderate and highly invasive tumors. Further analysis with SMR grouped in 10% intervals demonstrated an increase in overall survival for all groups, with a lower SMR extension to benefit nodular tumors.

Analysis of Globular Cells in Corneal Nerve Vortex.

PurposeLess was known about globular cells which were a type of dendritic cells (DCs) in cornea. We aimed to investigate the morphological and distribution characteristics of globular cells in corneal vortex and their clinical correlations with ocular surface.MethodsCase records of patients who underwent in vivo confocal microscopy (IVCM) were evaluated retrospectively. The morphology and distribution features of globular cells in cornea nerve vortex and their co-existence status with Langerhans cells (LCs) were analyzed. Data of ocular surface symptoms and signs were collected and their correlations with globular cells distribution patterns and dendritic forms were performed. Dry eye patients without LCs were treated with preservative-free artificial tears, while patients with LCs were treated with artificial tears and fluoromethalone until the activated LCs disappeared.ResultsA total of 836 eyes from 451 individuals were included. Three distribution patterns of globular cells in vortex were investigated, type 1 scattered globular cells (57.66%), type 2 large amounts of globular cells (≥50 cells) gathering in vortex and along some fixed vortex direction horizontally (13.52%) and type 3 no globular cells (28.83%). Their location and cell count altered slightly in the follow-ups but would not disappear. LCs could co-exist with globular cells and could fade after treatment. The type 2 distribution pattern was associated with older age (p = 0.000) and higher upper eyelid Meiboscore (p = 0.006). Dendritic globular cells had higher Meiboscore than Non-dendritic forms.ConclusionsGlobular cells had characteristic distribution patterns and biological features different from LCs. They were associated with long-term irritation of the meibomian gland dysfunction.

Combined Consideration of Tumor-Associated Immune Cell Density and Immune Checkpoint Expression in the Peritumoral Microenvironment for Prognostic Stratification of Non-Small-Cell Lung Cancer Patients.

Given the complexity and highly heterogeneous nature of the microenvironment and its effects on antitumor immunity and cancer immune evasion, the prognostic value of a single immune marker is limited. Here, we show how the integration of immune checkpoint molecule expression and tumor-associated immune cell distribution patterns can influence prognosis prediction in non-small-cell lung cancer (NSCLC) patients. We analyzed tissue microarray (TMA) data derived from multiplex immunohistochemistry results and measured the densities of tumor-infiltrating CD8+ and FOXP3+ immune cells and tumor cells (PanCK+), as well as the densities of programmed cell death 1 (PD-1)+ and programmed cell death ligand 1 (PD-L1)+ cells in the peritumor and intratumor subregions. We found a higher density of infiltrating CD8+ and FOXP3+ immune cells in the peritumoral compartment than in the intratumoral compartment. In addition, unsupervised hierarchical clustering analysis of these markers revealed that the combination of high CD8/FOXP3 expression, low PD-1 and PD-L1 immune checkpoint expression, and lack of epidermal growth factor receptor (EGFR) mutation could be a favorable predictive marker. On the other hand, based on the clustering analysis, low CD8/FOXP3 and immune checkpoint (PD-1 and PD-L1) expression might be a marker for patients who are likely to respond to strategies targeting regulatory T (Treg) cells. Furthermore, an immune risk score model was established based on multivariate Cox regression, and the risk score was determined to be an independent prognostic factor for NSCLC patients. These results indicate that the immune context is heterogeneous because of the complex interactions of different components and that using multiple factors in combination might be promising for predicting the prognosis of and stratifying NSCLC patients.