Cell Ratio Research Articles

NIMG-27. RADIOMETHYLOMICS FOR NON-INVASIVE PREDICTION OF TUMOR PHENOTYPES AND SURVIVAL IN GLIOBLASTOMA

Abstract BACKGROUND DNA methylation profiling has become a pivotal tool in neuro-oncology. However, little is known about the impact of methylation profiling on radiological imaging in IDH-wildtype glioblastoma (GBM). This study explored the potential of radiomics to non-invasively predict molecular characteristics based on the methylation profiling of GBM. METHODS This study included a multi-sampling cohort of 32 patients with 113 samples and a single-sampling cohort of 87 patients with 87 samples, which underwent genome-wide methylation analysis and were classified as GBM by the DKFZ/Heidelberg CNS tumor classifier. We performed two different methylation-based deconvolution analyses. Deconvolution 1 estimated the fractions of tumor subtypes (RTK_I, RTK_II, MES_TYP, and MES_ATYP) and cell types in the microenvironment. Deconvolution 2 inferred the abundance of malignant cell states (stem-like and differentiated cell components) and cell types in the microenvironment. We derived 6084 radiomic features from preoperative multi-parametric MRI in each patient. RESULTS In the multi-sampling cohort, Deconvolution 1 and Deconvolution 2 demonstrated that tumor subclass components and immune components exhibited heterogeneous distribution across samples within patients; however, the stem-like to differentiated cell ratio was preserved across samples within patients. In the entire cohort, patients with tumors exhibiting a high proportion of the stem-like component, defined as stem-like tumors, had significantly shorter overall survival. Stem-like tumors exhibited significantly higher levels of the RTK_I component and lower proportions of the RTK_II component compared to non-stem-like tumors. A machine learning model that utilizes support vector machines to classify stem-like tumors using radiomic features achieved a mean AUC of 0.71 (95% confidence interval: 0.56–0.85) in nested cross-validation. CONCLUSIONS We showed that radiomethylomics can be used to predict tumor composition and patient survival from preoperative MRI in GBM. Based on these promising results we will increase our power with additional samples to facilitate a personalized approach to GBM management.

Exploring the Role of Gut Vascular Barrier Proteins in HIV-Induced Mucosal Damage: A Comparative Study.

This study aims to compare intestinal mucosal damage and the expression levels of occludin, zonula occludens-1 (ZO-1), vascular endothelial (VE)-cadherin, β-catenin, and plasmalemma vesicle-associated protein (PLVAP) in the gut vascular barrier (GVB) among people living with HIV (PLWH), asymptomatic PLWH, and healthy volunteers (non-PLWH). Three groups were selected for the study: PLWH, asymptomatic PLWH, and healthy volunteers. Colonic mucosal tissue samples were collected via colonoscopy from all participants. Histological examination of the colonic mucosa was conducted using hematoxylin and eosin staining. The expression levels of occludin, ZO-1, VE-cadherin, β-catenin, and PLVAP were assessed using RT-qPCR, immunohistochemistry, and western blot analyses. Pathological scores of colonic mucosa in PLWH and asymptomatic PLWH were significantly higher than those in non-PLWH (p < .001 and p = .0056, respectively). CD4+ T cell counts in asymptomatic PLWH and non-PLWH were significantly higher than in PLWH (p < 0.05). The CD4+/CD8+ T cell ratio in non-PLWH significantly exceeded those in PLWH and asymptomatic PLWH (p < .05). Analysis of protein and mRNA expression revealed: (1) no statistically significant differences in PLVAP-mRNA expression across all groups (p > .05); (2) higher PLVAP protein levels in PLWH compared with asymptomatic PLWH and non-PLWH (p < .05), with no significant differences between asymptomatic PLWH and non-PLWH (p = .632); (3) significantly higher PLVAP expression in the colonic mucosa of PLWH and asymptomatic PLWH compared with non-PLWH (p = .034 and p = .011, respectively), with no significant differences between PLWH and asymptomatic PLWH (p > .999). ZO-1 expression was significantly lower in PLWH than in non-PLWH (p = .012), with no notable differences between asymptomatic PLWH and other groups. PLWH, compared with healthy controls, exhibit significant inflammatory changes in the intestinal mucosa. PLVAP expression serves as a potential indicator to assess the extent of GVB damage and disease progression in PLWH.

Separation of Lung Cancer Cells From Mixed Cell Samples Using Aptamer-Modified Magnetic Beads and Permalloy Micromagnets.

This study involved the design and fabrication of a microfluidic chip integrated with permalloy micromagnets. The device was used with aptamer-modified magnetic beads (MBs) of various sizes to successfully separate lung cancer cells from a mixture of other cells. The overall separation efficiency was evaluated based on the ratios of cells in the different outlets and inlets of the chip. The results showed efficiencies ranging from 43.4% to 50.2% for MB sizes between 1.36 and 4.50 µm. Interestingly, efficiency slightly decreased as the size of the MBs increased, contrary to predictions. Further examination revealed that larger MBs exerted gravitational force on the cell-bound MBs at low flow rates, causing the targets to settle before reaching the main microchannel region. This was attributed to fluidic resistance caused by a size mismatch between the inlet tube and the microfluidic conduit. An increase in cell accumulation at the inlet was observed with larger MB sizes due to gravity. Therefore, the definition of effective separation efficiency was revised to exclude the effect of cell accumulation at the inlet. Effective separation efficiencies were found to be 71.6%, 76.4%, and 79.4% for MB sizes of 1.36, 3.00, and 4.50µm, respectively. The study concluded that larger MBs interacted more with the magnetic force, resulting in better separation. However, cells with smaller MBs were more likely to evade the magnetic force. The investigation provides valuable insights into isolating lung cancer cells using this method, with the potential for clinical application in cancer diagnosis and treatment.

Clinical, pharmacology and in vivo studies of QingDai (indigo naturalis) promotes mucosal healing and symptom improvement in ulcerative colitis by regulating the AHR-Th17/Treg pathway.

Qingdai (QD), derived from various plant sources, is commonly used in traditional Chinese medicine for ulcerative colitis (UC) treatment. However, the clinical efficacy and mechanisms of orally administered QD remain unclear. This study aims to evaluate QD's efficacy in UC treatment and uncover its active components and mechanisms. A randomized controlled trial compared QD to Adisa, followed by UPLC-Q-TOF/MS and network pharmacology analyses to identify QD's core components and targets. In vivo experiments on a UC mouse model explored QD's impact on the AHR-Th17/Treg pathway using PCR, WB, ELISA, and flow cytometry. Results showed QD's efficacy in UC treatment, with mucosal healing and remission comparable to Adisa. UPLC-Q-TOF/MS identified 16 core components in mouse colon tissue, with network pharmacology revealing 67 targets, potentially involving the IL-17 signaling pathway and Th17 cell differentiation. QD and its components up-regulated AHR, CYP1A1, and Foxp3, while down-regulating RORγt. Additionally, QD modulated pro-inflammatory (IL-6, IL-17A) and anti-inflammatory (IL-10, TGF-β1) factors, and Treg/Th17 cell ratios in LPMC. Oral QD administration effectively promoted mucosal healing and improved UC symptoms, potentially through AHR-Th17/Treg pathway regulation.

Clinical and cytological characteristics of serous effusions in 69 cases of lymphoma patients.

To explore the value of cell morphology, immunophenotype, and gene alterations of serosal effusion in the diagnosis of lymphoma. Serosal effusion of 69 cases of lymphoma patients were collected, including 36 cases with malignant effusion and 33 cases with nonmalignant effusion. Ordinary cytology, liquid-based cytology, cellblock, and immunocytochemical staining were performed in each case, some cases were detected by fluorescence in situ hybridization for C-MYC, BCL2, and BCL6 gene translocations. T/B cell ratio in malignant and nonmalignant serosal effusions was analyzed and compared by flow cytometry (FCM)and immunohistochemical (IHC), respectively. The prognostic value of serous effusion in diffuse large B-cell lymphoma (DLBCL) was investigated and another 20 DLBCL cases without effusion were successively selected as control. The number of naive lymphocytes, apoptotic bodies, and mitotic figures were more common in malignant effusions compared with nonmalignant effusions (p < .01). The top three lymphomas in malignant effusion were DLBCL (19/36, 52.8%), mantle cell lymphoma (MCL) (4/36, 11.1%, 3 blastoid variant) and high-grade B-cell lymphoma (HGBL) (4/36, 11.1%). T/B cell ratio by FCM analysis ranged from 0.00 to 0.55 (mean 0.084) in malignant effusion, and 2.58 to 984.00 (mean 249.9) in nonmalignant effusion. The difference was significant (p = .017). The T/B cell ratio by IHC analysis ranged from 0.02 to 3.00 (mean 0.200) in malignant effusion, and 2.00-100.00 (mean 34.10) in nonmalignant effusion. The difference was significant (p = .017). In the effusions involving DLBCL, most effusions were present at the time of diagnosis (57.9%); single pleural effusions were more common (36.8%). The median overall survival times of patients with malignant effusion, nonmalignant effusion and DLBCL without serous effusion were 11, 17, and 23 months respectively (p = .04). Three patients of HGBL died, and the overall survival times were 5, 8, and 9 months, respectively. The cytomorphological characteristics combined with immunophenotype, FCM, gene rearrangement, and other tests can diagnose and classify patients with effusion as the first symptom. The T/B cell ratio is less than 1 by FCM or IHC suggesting a malignant serosal effusion. The presence of malignant effusion in DLBCL patients is an important clue for poor prognosis.

Cordycepin Enhances the Anticancer Efficacy of PD-L1 Blockade by Modulating the Tumor Microenvironment of Colon Cancer

BackgroundPD-L1 blockade has been found to be effective in treating multiple malignancies. Combined therapy is proposed to provide better therapeutic effects. Cordycepin, a prominent bioactive compound found in cordyceps, can inhibit the development of various cancers. PurposeThis study aimed to determine the efficacy of combined anti-PD-L1 antibody and cordycepin in tumor treatment. MethodsA single-cell RNA sequencing was used to analyze the mechanism of combined treatment. ResultsCombination therapy of anti-PD-L1 and cordycepin significantly inhibited tumor growth by regulating the T cell ratio and improving the function of CD8+T cells. Furthermore, cordycepin promoted the reprogramming of type-II macrophages into type-I macrophages, a process confirmed through flow cytometry analysis of the underlying mechanism. ConclusionOur findings demonstrate that the combination of anti-PD-L1 and cordycepin effectively suppressed tumor growth by regulating the proportion of T cells and reprograming type-II macrophages.

Phytochemical examination of Cistus laurifolius Extract and its Impact on Cytotoxicity, Apoptosis and Oxidative Stress in Colorectal and Breast Cancer Cell Lines

IntroductionColon and breast cancer are the most common types of cancer worldwide. This study investigatesd the anticancer properties of Cistus laurifolius L. leaf extract on breast cancer (MCF-7) and human colon cancer (Caco-2) cell lines. MethodsThe research involved meticulous collection, drying and processing of C. laurifolius leaves to extract bioactive compounds, subsequently analyzed for phenolic content using advanced LC-MS/MS technology. Cell viability of the extract on MCF-7 and Caco-2 cells was demonstrated by MTT test. Levels of critical apoptotic markers (Bad, Bax/Bcl-2, Bax/Bcl-xl, Caspase-9) and Total Antioxidant Capacity (TAC) and Total Oxidant Capacity (TOC), which affect the antioxidant system, were evaluated by the ELISA method. Identification of phenolic compounds, including quercetin and rutin, through target prediction analysis enriches our understanding of bioactive molecules. ResultsThe results of the study showed that C. laurifolius extract inhibited cell proliferation time and dose-dependent on Caco-2 and MCF-7 cells (P<0.05). TAC, Bax/Bcl-2 and Bax/Bcl-xl ratios in MCF-7 and Caco-2 cells increased in a dose-dependent manner compared to the control group. In MCF-7 cells, TAC (p<0.05; p<0.01), Bax/Bcl-2 (p<0.001; p<0.0001) and Bax-Bcl-xl (p<0.01) ratios increased at 24 hours compared to the control group. In Caco-2 cells, TAC (p<0.001), Bax/Bcl-2 ratios increased at 48 hours (p<0.05), while Bax-Bcl-xl ratios decreased (p<0.01; p<0.001) compared to the control group. ConclusionC. laurifolius leaf extracts emerge as a promising anticancer candidate, hindering cell proliferation and inducing apoptosis in colon and breast cancer cells. The classification of bioactive molecules may facilitate further clinical therapeutic interventions targeting colon and breast cancer.

Luteolin prevents cadmium-induced PC12 cell death by suppressing the Akt/mTOR signaling pathway.

Cadmium (Cd) is an environmental pollutant that can cause neurodegenerative disorders. Luteolin (Lut) is a natural flavonoid compound. However, whether Lut protects against Cd-induced nerve cell death remains unclear. In the present study, PC12 cells were used to investigate the neuroprotective effect of Lut against Cd poisoning. Changes in cell viability, apoptosis, B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein expression, and protein kinase B (Akt)/mammalian target of rapamycin (mTOR) pathway activity were analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Hoechst 33258 staining, flow cytometry, and western blotting. Lut markedly attenuated the Cd-induced reduction in cell viability, nuclear fragmentation, condensation, and the decrease in the Bcl-2/Bcl-2-associated X protein ratio in PC12 cells. Furthermore, Lut blocked the Cd-mediated activation of the Akt/mTOR signaling pathway. Moreover, inhibition of the Akt/mTOR signaling pathway with LY294002 (a PI3K inhibitor) enhanced the protective effect of Lut against Cd-induced cell death by suppressing Cd-induced activation of Akt, mTOR, and eukaryotic initiation factor 4E binding protein 1. The results showed that Lut prevented Cd-induced cell death partly by blocking the Akt/mTOR signaling pathway. Lut may be a potential agent for preventing Cd-induced nerve cell damage and neurodegenerative diseases.

LAG-3 Expression, γδ-T cell/MHC-I Interactions and Prognosis in Merkel Cell Carcinoma

Merkel Cell Carcinoma (MCC) is an aggressive cutaneous malignancy with a poor prognosis. One of the major mechanisms of immune evasion in MCC involves downregulation of MHC-I. Anti-PD-1/PD-L1 checkpoint inhibitors (CKIs) have revolutionized treatment for MCC, producing objective responses in ∼50% of patients, and are now standard of care; however, a substantial proportion of patients either fail to respond or develop resistance to CKIs. Given these recent successes, identification of other targetable immune checkpoints in the MCC tumor microenvironment (TME) is of great interest. Additionally, γ-delta (γδ) T cells may play critical roles in the response to MHC-I deficient cancers; therefore, evaluating γδ-T cells as a prognostic biomarker is warranted. We characterized the expression of PD-L1, PD-1, CD3, CD8, LAG-3, MHC-I, and γδ-T cells by IHC in a pre-immunotherapy retrospective cohort of 54 cases of MCC, and quantified expression levels and marker density via HALO. The increased density of LAG-3 and γδ-T cells correlated with other markers of an inflamed TME, with significant positive associations across all six markers (p<.002). Reflective of their putative role in the response to MHC-I suppressed cancers, cases with low HLA-I density showed a trend towards a higher ratio of γδ-T cells:CD3+ T cells (Spearman's r=-0.1582, p=0.21). Importantly, high CD3 density (HR=0.23, p=0.002), LAG-3 density (HR=0.47, p=0.037), γδ-T cell density (HR=0.26, p=0.02), and CD8 density (HR=0.27, p=0.03) showed associations with improved progression-free survival. Conditional tree analysis demonstrated that high CD8 and TCRδ expression were non-significant predictors of improved PFS and OS. Overall, LAG-3 is expressed in MCC infiltrates and is prognostic in pre-immunotherapy MCC, suggesting a potential role for LAG-3 inhibition in MCC. Additionally, CD8 and γδ-T cells may play a critical role in the response to MCC, and γδ-T cell density may represent a novel biomarker in MCC.

A machine-learning-based algorithm for bone marrow cell differential counting

BackgroundDifferential counting (DC) of different cell types in bone marrow (BM) aspiration smears is crucial for diagnosing hematological diseases. However, a clinically applicable method for automatic DC has yet to be developed. ObjectiveThis study developed and validated an artificial intelligence (AI)-based algorithm for identifying and classifying nucleated cells in BM smears. MethodsIn the development phase, a mask region–based convolutional neural network (Mask R-CNN)-based AI model was trained to detect and classify individual BM cells. We used a large data set of expert-annotated images representing a variety of disease categories. The BM slides were stained with Liu’s stain or Wright–Giemsa stain. Consensus meetings were held to ensure experts from different institutes applied consistent criteria in classifying cells. Subsequently, the performance of the AI algorithm in identifying cell images and determining cell ratios was evaluated using a multinational clinical dataset. ResultsThe AI model was trained on 542 slides (85.1 % stained with Liu’s stain and 14.9 % with Wright–Giemsa stain) containing 597,222 annotated cells. It achieved an accuracy of 0.94 for the testing dataset containing 26,170 cells. The performance of the AI model was further validated using another multinational real-world dataset (data obtained from three centers in Taiwan and one in the United States) comprising 200,639 cells. The AI model achieved an accuracy of 0.881 in classifying individual cells and demonstrated high precision in classifying blasts (0.927), bands and polymorphonuclear neutrophils (0.955), plasma cells (0.930), and lymphocytes (0.789). When the differential counting percentage of each cell type was assessed, a strong correlation (ρ > 0.8) between the AI and manual methods was observed for most cell categories. ConclusionsIn this study, an AI algorithm was developed and clinically validated using large, multinational datasets. Our algorithm can locate and classify BM cells simultaneously and has potential clinical applicability for automating BM differential counting.

Association of innate versus specific immunity with heart failure incidence: a prospective study.

Immune disorders are key heart failure (HF) triggers, but little is known about whether the status of immunity affects the incidence of HF. To explore this, we used blood cell counts and derived ratios to investigate the association between immunity status markers and HF incidence. The number and proportion of peripheral blood leucocytes in a physiological state are related to the body's immune status. Neutrophils, monocytes, SII (systemic immune-inflammatory index), NLR (neutrophil-to-lymphocyte ratio), and PLR (platelet-to-lymphocyte ratio) serve as innate immunity status markers, while lymphocytes and LMR (lymphocyte-to-monocyte ratio) serve as specific immunity status markers. 330 362 UK Biobank (UKB) participants were finally examined. Cox proportional hazard models were used to explore the relationship between immunity status markers and HF incidence. Flexible parametric survival models were used to capture time-varying relationships between blood cell ratios and HRs for HF. Subgroup analyses were conducted by age, sex, and body mass index. Finally, sensitivity analyses were performed to validate the results. During a median follow-up of 14.1 years, 9611 (2.9%) participants developed HF. Neutrophils, monocytes, SII, and NLR were positively associated with HF incidence, with fully adjusted per SD increment HR (95% CI) of 1.20 (1.17 to 1.22), 1.09 (1.07 to 1.12), 1.12 (1.10 to 1.14), and 1.16 (1.14 to 1.18), respectively. Platelets, lymphocytes, and LMR were inversely correlated with HF incidence, with fully adjusted per SD increment HR (95% CI) of 0.97 (0.95 to 1.00), 0.97 (0.95 to 0.99), and 0.90 (0.88 to 0.92), respectively. The innate immunity status markers were positively associated with HF incidence, while specific immunity status markers exhibited an inverse association, offering novel insights for HF prediction and intervention.

Prognostic values of combined ratios of white blood cells in glioma: a systematic review and meta-analysis.

Gliomas, the most prevalent type of neurological tumor, pose a challenging prognosis for patients. Recent studies have underscored the importance of inflammatory markers such as the neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), and monocyte/lymphocyte ratio (MLR) in predicting the prognosis of gliomas. We undertook a thorough meta-analysis to elucidate the role of these inflammatory markers in forecasting the prognosis of glioma patients. We extracted hazard ratios (HR) and their corresponding 95% confidence intervals (95% CI) from each study for analysis. To assess heterogeneity and identify influential studies, we conducted sensitivity analysis. Subgroup analysis was performed to investigate sources of heterogeneity, and we employed Egger's test to evaluate publication bias in the meta-analysis. Higher NLR levels were associated with shorter overall survival (HR = 1.46, 95% CI: 1.33-1.60) and progression-free survival (HR = 1.24, 95% CI: 1.04-1.48). There was no significant correlation between PLR levels and overall survival (HR = 1.01, 95% CI: 1.00-1.01) or progression-free survival (HR = 1.00, 95% CI: 0.98-1.02) in glioma patients. Elevated MLR levels were associated with decreased overall survival in glioma patients (HR = 1.78, 95% CI: 1.36-2.34). SII levels did not show any significant association with overall or progression-free survival in glioma patients (HR = 1.00, 95% CI: 0.99-1.01).In the sensitivity analysis, two studies potentially contributed to the instability. Subgroup analyses showed patient population and area were identified as potential sources of heterogeneity. Egger's test showed that there was publication bias in the relationship between NLR and PLR and overall survival (P < 0.05).All randomized controlled models, except for these, were not affected by publication bias. NLR and MLR are two reliable indicators of inflammation in the prognosis of glioma patients; PLR and SII do not have significant value in the prognosis of glioma patients.

Effect of Voluntary Wheel Running on Anxiety- and Depression-Like Behaviors in Fluoride-Exposed Mice.

Fluoride, an environmental toxicant, could induce endoplasmic reticulum stress (ERS) in neuronal cells ultimately leading to apoptosis and emotional dysfunction. Meanwhile, voluntary wheel running contributes to mitigate anxiety and depression. Our investigation aimed to study the effect of voluntary wheel running on anxiety- and depression-like behaviors in fluoride-exposure mice. The results showed that exposure to 100 mg/L sodium fluoride (NaF) for 6 months can induce anxiety- and depression-like behavior in mice. Fluorosis mice subjected to voluntary wheel running have less anxiety- and depression-like behaviors. Nissl and TUNEL staining demonstrated that fluoride led to a reduced proportion of Nissl body area in the cerebral cortex and an increased apoptotic ratio of nerve cells in the cerebral cortex. In contrast, these pathologic damages were improved in voluntary wheel running mice exposed to NaF. Moreover, the expressions of mRNA in the cerebral cortex GABA, GAD65, GAD67, DR, vGLU, 5-HT1A, BDNF, NMDAR1, and Bcl2 were downregulated and the levels of c-fos, GRP78, PERK, eIF2α, CHOP, Caspase-12, and Caspase-3 mRNA were upregulated in mice exposed to fluoride. NaF treatment had increased the PERK, ATF6, IRE1, p-eIF2α, and Caspase-3 protein levels and reduced the expressions of proteins, including GAD67, VGAT, BDNF, NMDAR1, PSD95, and SYN. By contrast, fluorosis mice subjected to voluntary wheel running enhanced the expression of GAD65, GAD67, VGAT, and neuroplasticity-related proteins in mice and inhibited the PERK-CHOP pathway. It is worth noting that the correlation between the amount of exercise and the behavioral indicators as well as neurotransmitter levels was found. In conclusion, voluntary wheel running inhibits the fluoride-induced ERS and GRP78 expression through the PERK-CHOP pathway and plays an anti-apoptotic role, ultimately ameliorating emotional dysfunction in NaF-exposed mice.

Additive manufactured 3D re-entrant auxetic structures for enhanced impact resistance

Abstract This study presents a novel exploration of the geometric parameters within a 3D re-entrant auxetic lattice structure, specifically focusing on their unique impact energy absorption properties, which were systematically evaluated through drop weight impactor testing. Each lattice configuration was additively manufactured using stereolithography, allowing for precise control over strut thickness (t), re-entrant angle (θ), and the aspect ratio (h/l) of unit cells during both low and high energy impact scenarios. This study found that the overall auxetic behavior is predominantly controlled by the aspect ratio of the cell ribs, while the modulus is governed by rib thickness. A finite element model was subsequently developed to simulate the experimental impact loading conditions and was used to examine a wider range of parameters that were not experimentally tested. The simulated dynamic test results displayed the deformation trends and changes to the Poisson's ratio. Among the studied parameters, experimental results highlighted that a lattice structure with t = 1.6 mm, θ = 65°, and a h/l ratio = 1.8 exhibited the highest specific energy absorption (SEA) under uniaxial impact deformation with 5 Joules of impact energy. Conversely, when employing 20 Joules of impact energy revealed the greatest SEA at t = 1.0 mm, θ = 65°, and an h/l ratio of 2.2. The results demonstrate unique deformation mechanism of auxetic structures under impact loading and the capacity to adapt the 3D re-entrant lattice structure for applications requiring tailored impact energy absorption.

Characterization of white blood cell ratios in South American camelids presented at a veterinary teaching hospital

White blood cell (WBC) ratios are used as diagnostic markers for various inflammatory or tumor diseases as well as stress in a broad range of species. The aim of this work was to provide data on five WBC ratios (neutrophil-to-lymphocyte ratio [NLR], band neutrophil-to-lymphocyte ratio [BLR], band neutrophil-to-neutrophil-to-lymphocyte ratio [BNLR], band neutrophil-to-neutrophil ratio [BNR] and lymphocyte-to-monocyte ratio [LMR]) in South American camelids (SAC) and characterize their association with demographic and important diagnostic parameters. Medical records of 307 SAC (275 alpacas, 32 llamas) that were presented at a veterinary teaching hospital were evaluated retrospectively. WBC ratios were calculated based on hematologic results of the initial blood samples. The influence of species, sex, age, body condition score, WBC count, and anemia on those ratios was investigated using descriptive statistics and generalized linear models. NLR, BLR and LMR were found to be significantly influenced by age and WBC count. Associations of individual WBC ratios with species, nutritional status or an anemic condition could be detected. NLR was 4.32; 2.31–7.81 (median; IQR), BLR 0.24; 0.07–0.87, BNLR 3.66 × 10–3; 1.17 × 10–3 − 14.20 × 10–3, BNR 0.06; 0.02–0.15 and LMR was 7; 3.54–14.67. Our data might serve as a basis for further studies on WBC ratios in SAC. The animals in this study showed a variety of underlying diseases. It should hence be noted that these values are orientation values and provide a representative overview of conditions in a clinic, but are not suitable as reference values for healthy animals.

Platelet-large cell ratio predicts no-reflow in patients undergoing primary coronary angioplasty for ST-segment elevation myocardial infarction

Abstract Background Increased platelet indices have been shown to reflect higher degree of platelet activation and have been related to poorer short- and long-term outcomes in patients presenting with ST-segment elevation myocardial infarction (STEMI). Purpose We aimed to evaluate the ability of a validated marker of platelet activation (i.e., the platelet-large cell ratio [P-LCR]) to predict the occurrence of no-reflow in patients undergoing primary percutaneous coronary intervention (pPCI) for STEMI. The ability of P-LCR to predict other markers of thrombotic charge (i.e., use of glycoprotein IIb/IIIa inhibitors and/or thrombus aspiration) was also investigated. Methods Use of glycoprotein IIb/IIIa inhibitors and/or thrombus aspiration, occurrence of the ‘no-reflow’ phenomenon and P-LCR values on admission were recorded in 836 consecutive patients treated by pPCI for STEMI. Patients’ demographic data and symptom onset-to-cardiac catheterization laboratory time intervals were also recorded. Correlations between P-LCR values and the presence of thrombotic charge markers were evaluated. Results No-reflow occurred in 124 (14.8%) of the study patients; glycoprotein IIb/IIIa inhibitors and thrombus aspiration were used in 228 (27.3%) and in 289 (34.5%) of patients, respectively. Mean P-LCR values were 27.1 ± 7.6%. P-LCR values on admission were significantly higher in patients who presented no-reflow than in those who did not (29.1 ± 5.6% vs. 25.3 ± 5.8%, p = 0.03). Similarly, P-LCR were significantly higher in patients in whom glycoprotein IIb/IIIa inhibitors (29.5 ± 6.1% vs. 25.2 ± 5.3%, p = 0.04) and/or thrombus aspiration (28.4 ± 5.7% vs. 24.8 ± 5.4%, p = 0.03) were used. A P-LCR value ≥26.9% predicted the occurrence of no-reflow with 70.9% sensitivity and 55.4% specificity (p = 0.04). Conclusions In the present cohort, P-LCR values on admission predicted the presence of thrombotic charge markers in patients undergoing pPCI for STEMI. Evaluating P-LCR on admission could help identify patients at increased risk of no-reflow, in whom more intensive antiplatelet strategies may be of interest.

From bite to brain: Neuro-immune interactions in food allergy.

Immunoglobulin E (IgE)-mediated food allergies are reported to affect around 3.5% of children and 2.4% of adults, with symptoms varying in range and severity. While being the gold standard for diagnosis, oral food challenges are burdensome, and diagnostic tools based on specific IgE can be flawed. Furthering our understanding of the mechanisms behind food allergy onset, severity and persistence could help reveal immune profiles associated with the disease, to ultimately aid in diagnosis. Alterations to cytokine levels and immune cell ratios have been identified, though further research is needed to fully capture the heterogenous nature of food allergy. Moreover, the existence of such immune alterations also raises the question of potential wider systemic effects. For example, recent research has emphasised the existence and impact of neuro-immune interactions and implicated behavioural and neurological changes associated with food allergy. This review will provide an overview of such food allergy-driven neuro-immune interactions, with the aim of emphasising the importance of furthering our understanding of the immune mechanisms underlying IgE-mediated food allergy.

In Vitro Assessment of Thermo-Responsive Scaffold as a 3D Synthetic Matrix for CAR-T Potency Testing Against Glioblastoma Spheroids.

Chimeric antigen receptor (CAR) T cell immunotherapy has demonstrated exceptional efficacy against hematological malignancies, but notably less against solid tumors. To overcome this limitation, it is critical to investigate antitumor CAR-T cell potency in synthetic 3D microenvironments that can simulate the physical barriers presented by solid tumors. The overall goal of this study was the preliminary assessment of a synthetic thermo-responsive material as a substrate for invitro co-cultures of anti-disialoganglioside (GD2) CAR-T cells and patient-derived glioblastoma (GBM) spheroids. Independent co-culture experiments demonstrated that the encapsulation process did not adversely affect the cell cycle progression of glioma stem cells (GSCs) or CAR-T cells. GSC spheroids grew over time within the terpolymer scaffold, when seeded in the same ratio as the suspension control. Co-cultures of CAR-T cells in suspension with hydrogel-encapsulated GSC spheroids demonstrated that CAR-T cells could migrate through the hydrogel and target the encapsulated GSC spheroids. CAR-T cells killed approximately 80% of encapsulated GSCs, while maintaining effective CD4:CD8 T cell ratios and secreting inflammatory cytokines after interacting with GD2-expressing GSCs. Importantly, the scaffolds also facilitated cell harvesting for downstream cellular analysis. This study demonstrated that a synthetic 3D terpolymer hydrogel can serve as an artificial scaffold to investigate cellular immunotherapeutic potency against solid tumors.

Aquificae overcomes competition by archaeal thermophiles, and crowding by bacterial mesophiles, to dominate the boiling vent-water of a Trans-Himalayan sulfur-borax spring.

Trans-Himalayan hot spring waters rich in boron, chlorine, sodium and sulfur (but poor in calcium and silicon) are known based on PCR-amplified 16S rRNA gene sequence data to harbor high diversities of infiltrating bacterial mesophiles. Yet, little is known about the community structure and functions, primary productivity, mutual interactions, and thermal adaptations of the microorganisms present in the steaming waters discharged by these geochemically peculiar spring systems. We revealed these aspects of a bacteria-dominated microbiome (microbial cell density ~8.5 × 104 mL-1; live:dead cell ratio 1.7) thriving in the boiling (85°C) fluid vented by a sulfur-borax spring called Lotus Pond, situated at 4436 m above the mean sea-level, in the Puga valley of eastern Ladakh, on the Changthang plateau. Assembly, annotation, and population-binning of >15-GB metagenomic sequence illuminated the numeral predominance of Aquificae. While members of this phylum accounted for 80% of all 16S rRNA-encoding reads within the metagenomic dataset, 14% of such reads were attributed to Proteobacteria. Post assembly, only 25% of all protein-coding genes identified were attributable to Aquificae, whereas 41% was ascribed to Proteobacteria. Annotation of metagenomic reads encoding 16S rRNAs, and/or PCR-amplified 16S rRNA genes, identified 163 bacterial genera, out of which 66 had been detected in past investigations of Lotus Pond's vent-water via 16S amplicon sequencing. Among these 66, Fervidobacterium, Halomonas, Hydrogenobacter, Paracoccus, Sulfurihydrogenibium, Tepidimonas, Thermus and Thiofaba (or their close phylogenomic relatives) were presently detected as metagenome-assembled genomes (MAGs). Remarkably, the Hydrogenobacter related MAG alone accounted for ~56% of the entire metagenome, even though only 15 out of the 66 genera consistently present in Lotus Pond's vent-water have strains growing in the laboratory at >45°C, reflecting the continued existence of the mesophiles in the ecosystem. Furthermore, the metagenome was replete with genes crucial for thermal adaptation in the context of Lotus Pond's geochemistry and topography. In terms of sequence similarity, a majority of those genes were attributable to phylogenetic relatives of mesophilic bacteria, while functionally they rendered functions such as encoding heat shock proteins, molecular chaperones, and chaperonin complexes; proteins controlling/modulating/inhibiting DNA gyrase; universal stress proteins; methionine sulfoxide reductases; fatty acid desaturases; different toxin-antitoxin systems; enzymes protecting against oxidative damage; proteins conferring flagellar structure/function, chemotaxis, cell adhesion/aggregation, biofilm formation, and quorum sensing. The Lotus Pond Aquificae not only dominated the microbiome numerically but also acted potentially as the main primary producers of the ecosystem, with chemolithotrophic sulfur oxidation (Sox) being the fundamental bioenergetic mechanism, and reductive tricarboxylic acid (rTCA) cycle the predominant carbon fixation pathway. The Lotus Pond metagenome contained several genes directly or indirectly related to virulence functions, biosynthesis of secondary metabolites including antibiotics, antibiotic resistance, and multi-drug efflux pumping. A large proportion of these genes being attributable to Aquificae, and Proteobacteria (very few were ascribed to Archaea), it could be worth exploring in the future whether antibiosis helped the Aquificae overcome niche overlap with other thermophiles (especially those belonging to Archaea), besides exacerbating the bioenergetic costs of thermal endurance for the mesophilic intruders of the ecosystem.

Benzylpiperazinyl derivatives of alepterolic acid: Synthesis and cytotoxic evaluation.

Natural products play a significant role in the development of modern drugs. Alepterolic acid, a labdane-type diterpenoid firstly isolated from Aleuritopteris argentea (Gmél.) Fée, has been identified as a valuable template for the synthesis of potent anticancer agents by structural modification. In this study, a series of new derivatives was obtained by coupling alepterolic acid with benzylpiperazines. It was found that (3,4-dichlorobenzyl)piperazinyl alepterolic acid (compound 6p) displayed the most toxic against MCF-7 cell line, with IC50 value of 8.31±0.67 μM. Further investigations demonstrated that compound 6p induced morphological changes in MCF-7 cells, inhibited proliferation in a time- and dose-dependent manner. Furthermore, western blot analysis revealed that compound 6p induced a significant increase in cleaved caspase-9, cleaved caspase-3, cleaved poly (ADP-ribose) polymerase (PARP) and Bax/Bcl2 ratio in MCF-7 cells. All of these results confirmed that compound 6p induced endogenous apoptosis in MCF-7 cells. Conclusively, the findings suggest that the incorporation of benzylpiperazine to alepterolic acid represents a promising approach for the discovery of new drug candidates.