Imaging flow cytometry (ImFC) represents a significant technological advancement in the field of cytometry, effectively merging the high-throughput capabilities of flow analysis with the image-based features of microscopy. In this comprehensive review article, we take a historical perspective and delve into the evolution of this methodology, presenting its past, exploring current state-of-the-art and forecasting potential future advancements. The inception of ImFC arose from the innovative combination of a flow cytometer's hydraulics with the technology of an advanced camera. This synergistic coupling facilitated the morphological analysis of cell populations at a high-throughput scale, effectively evolving the landscape of cytometry. However, the application of ImFC faced several challenges. Among the most prominent ones, is the development and implementation of software systems that can reliably and efficiently handle the robust data acquisition and analysis inherent in ImFC. The scale and complexity of the data generated by ImFC necessitates the creation of novel analytical tools that can effectively manage and interpret this data, thus allowing us to unlock the full potential of ImFC. Interestingly, artificial intelligence (AI) algorithms have begun to be implemented on ImFC, offering a promise for enhancing analytical capabilities. The adaptability and learning capacity of AI may prove to be essential in knowledge mining from high-dimensional data produced by ImFC, potentially enabling more accurate analyses. Looking forward, we project that ImFC may become an indispensable tool, not only in research laboratories but also in clinical settings. Given the unique combination of high-throughput cytometry and detailed imaging offered by ImFC, we foresee a critical role for this technology in the next generation of scientific research and diagnostics. As such, we encourage both current and future scientists to consider the integration of ImFC as an addition to their research toolkit and clinical diagnostic routine.

Using a randomized, crossover approach, cyclists (N = 20, overnight fasted) engaged in three 75-km time trials while ingesting water (WAT) or carbohydrate (0.2 g/kg every 15 minutes) from bananas (BAN) or a 6% sugar beverage (SUG). Blood samples were collected pre-exercise and 0 h-, 1.5 h-, and 21 h-post-exercise, and analyzed for NK cytotoxicity activity (NKCA) using pure NK cell populations. The two carbohydrate trials (BAN, SUG) compared to WAT were associated with higher post-exercise glucose, and lower cortisol, total blood leukocyte, neutrophil, and NK cell counts (interaction effects, P < 0.001). The immediate post-exercise increase in NK cell counts was higher in WAT (78%) compared to BAN (32%) and SUG (15%) trials (P ≤ 0.017). The 1.5 h post-exercise decrease in NK cell counts did not differ after WAT (−46%), BAN (−46%), and SUG (−51%) trials. The pattern of change in post-exercise NKCA differed between trials (P < 0.001). The 1.5 h post-exercise decreases in NKCA were 23%, 29%, and 33% in the WAT, BAN, and SUG trials, respectively, but trial contrasts did not differ significantly. Carbohydrate ingestion from BAN or SUG attenuated immediate-post-exercise increases in leukocyte, neutrophil, and NK cell counts, but did not counter the 1.5-h decreases in NK cell counts and NKCA.

C-C motif chemokine receptor 1 (CCR1/CD191) is a member of G-protein-coupled receptors, and is expressed on myeloid cells, such as neutrophils and macrophages. Because the CCR1 signaling promotes tumor expansion in the tumor microenvironment (TME), the modification of TME is an effective strategy for cancer therapy. Although CCR1 is an attractive target for solid tumors and hematological malignancies, anti-CCR1 therapeutic agents have not been approved. Here, we established a novel anti-mouse CCR1 (mCCR1) monoclonal antibody (mAb), C₁Mab-6 (rat IgG_2b, kappa), using the Cell-Based Immunization and Screening (CIBS) method. Flow cytometry and Western blot analyses showed that C₁Mab-6 recognizes mCCR1 specifically. The dissociation constant of C₁Mab-6 for mCCR1-overexpressed Chinese hamster ovary-K1 was determined as 3.9×10^-9 M, indicating that C₁Mab-6 possesses a high affinity to mCCR1. These results suggest that C₁Mab-6 could be a useful tool for targeting CCR1 in preclinical mouse models.

The C-X-C motif chemokine receptor-1 (CXCR1) is a rhodopsin-like G-protein-coupled receptor, expressed on the cell surface of immune cells and tumors. CXCR1 interacts with some C-X-C chemokines, such as CXCL6, CXCL7, and CXCL8/interleukin-8, which are produced by various cells. Since CXCR1 is involved in several diseases including tumors and diabetes mellitus, drugs targeting CXCR1 has been developed. Therefore, the development of sensitive monoclonal antibodies (mAbs) for CXCR1 has been desired for the diagnosis and treatment. This study established a novel anti-mouse CXCR1 (mCXCR1) mAb; Cx₁Mab-1 (rat IgG₁, kappa), using the Cell-Based Immunization and Screening (CBIS) method. Cx₁Mab-1 reacted with mCXCR1-overexpressed Chinese hamster ovary-K1 (CHO/mCXCR1) and mCXCR1-overexpressed LN229 glioblastoma (LN229/mCXCR1) in flow cytometry. Cx₁Mab-1 demonstrated a high binding affinity for CHO/mCXCR1 and LN229/mCXCR1 with a dissociation constant of 2.6×10^-9 M and 2.1×10^-8 M, respectively. Furthermore, Cx₁Mab-1 could detect mCXCR1 by western blot analysis. These results indicated that Cx₁Mab-1 is useful for detecting mCXCR1, and provides a possibility for targeting mCXCR1-expressing cells in vivo experiments.

Immunological aging type definition requires establishing reference intervals from the distribution of immunosenescence biomarkers conditional on age. For 1,605 individuals (18–97 years), we determined the comprehensive IMMmune Age indeX IMMAX from flow cytometry-based blood cell sub-populations, and identified age-specific centiles by fitting generalized additive models for location, scale and shape. The centiles were uncorrelated with age, and facilitated the categorization of individuals as immunologically slow or fast aging types. Using its 50th percentile as reference, we rescaled IMMAX to equivalent years-of-life (EYOL), and computed the immunological age gap as difference between EYOL and chronological age. Applied to preliminary baseline and follow-up measurements from 53 participants of the Dortmund Vital Study (Clinical-Trials.gov Identifier: NCT05155397), the averaged changes in IMMAX and EYOL conformed to the 5-year follow-up period, whereas no significant changes occurred concerning IMMAX centiles and age gap. This suggested that the participants immunologically adapted to aging, and kept their relative positions within the cohort. Sex was non-significant. Methodical comparisons indicated that future confirmatory analyses with the completed follow-up examinations could rely on percentile curves estimated by simple linear quantile regression, while the selection of the immunosenescence biomarker will greatly influence the outcome, with IMMAX representing the preferable choice.

CC chemokine receptor 6 (CCR6) is a member of the G protein-coupled receptor (GPCR) family that is highly expressed in B lymphocytes, effector and memory T cells, regulatory T cells, and immature dendritic cells. CCR6 has been revealed to have important functions in many pathological conditions, such as cancer, intestinal bowel disease, psoriasis, and autoimmune diseases. The only CCR6 chemokine ligand, CC motif chemokine ligand 20 (CCL20), is also involved in pathogenesis by interacting with CCR6. The CCL20/CCR6 axis is drawing attention as an attractive therapeutic target for various diseases. In this study, we developed novel monoclonal antibodies (mAbs) against human CCR6 (hCCR6) using the peptide immunization method, which are applicable to flow cytometry and immunohistochemistry. The established anti-hCCR6 mAb, clone C6Mab-19 (mouse IgG1, kappa), reacted with hCCR6-overexpressed Chinese hamster ovary-K1 (CHO/hCCR6), HepG2 (human liver carcinoma), and HuH-7 (human differentiated hepatoma) cells in flow cytometry. The dissociation constant (KD) of C6Mab-19 was determined as 3.0 × 10−10 M for CHO/hCCR6, 6.9 × 10−10 M for HepG2, and 1.8 × 10−10 M for HuH-7. Thus, C6Mab-19 could bind to exogenously and endogenously expressed hCCR6 with extremely high affinity. Furthermore, C6Mab-19 could stain formalin-fixed paraffin-embedded lymph node tissues from a patient with non-Hodgkin lymphoma by immunohistochemistry. Therefore, C6Mab-19 is suitable for detecting hCCR6-expressing cells and tissues, and could be useful for pathological analysis and diagnosis.

Immune Checkpoint Inhibitors are monoclonal antibodies that are used to treat over one in three cancer patients. While they have changed the natural history of disease, prolonging life and preserving quality of life, they are highly active in less than 40% of patients, even in the most responsive malignancies such as melanoma, and cause significant autoimmune side effects. Licenced biomarkers include tumour Programmed Death Ligand 1 expression by immunohistochemistry, microsatellite instability, and Tumour Mutational Burden, none of which are particularly sensitive or specific. Emerging tumour and immune tissue biomarkers such as novel immunohistochemistry scores, tumour, stromal and immune cell gene expression profiling, and liquid biomarkers such as systemic inflammatory markers, kynurenine/tryptophan ratio, circulating immune cells, cytokines and DNA are discussed in this review. We also examine the influence of the faecal microbiome on treatment outcome and its use as a biomarker of response and toxicity.

Overexpression of human epidermal growth factor receptor 2 (HER2) in breast and gastric cancers is an important target for monoclonal antibody (mAb) therapy. All therapeutic mAbs, including anti-HER2 mAbs, exhibit adverse effects probably due to the recognition of antigens expressed in normal cells. Therefore, tumor-selective or specific mAbs can be beneficial in reducing the adverse effects. In this study, we established a novel cancer-specific anti-HER2 antibody, named H₂Mab-250/H₂CasMab-2 (IgG₁, kappa). H₂Mab-250 reacted with HER2-positive breast cancer BT-474 and SK-BR-3 cells. Importantly, H₂Mab-250 did not react with non-transformed normal epithelial cells (HaCaT and MCF 10A) and immortalized normal epithelial cells in flow cytometry. In contrast, most anti-HER2 mAbs including H₂Mab-119 (IgG₁, kappa) reacted with both cancer and normal epithelial cells. Furthermore, a core-fucose deleted IgG_2a-type H₂Mab-250 (H₂Mab-250-mG_2a-f) could trigger the antibody-dependent cellular cytotoxicity activity to BT-474, but not to HaCaT cells. Furthermore, H₂Mab-250-mG_2a-f exhibited an in vivo antitumor effect against BT-474 xenograft. Immunohistochemical analysis demonstrated that H₂Mab-250 possesses much higher reactivity to the HER2-positive breast cancer tissues compared to H₂Mab-119, and did not react with normal tissues, including heart, breast, stomach, lung, colon, kidney, and esophagus. The epitope mapping demonstrated that the Trp614 of HER2 domain IV mainly contributes to the recognition by H₂Mab-250. H₂Mab-250 could contribute to the development of chimeric antigen receptor-T or antibody-drug conjugates without adverse effects for breast cancer therapy.

In the current study, we assessed the effects of conventional and Ultra High Dose Rate (UHDR) irradiation on behavioral and cognitive performance one month following exposure and assessed whether these effects were associated with alterations in the number of immune cells in the hippocampus using flow cytometry. Conventional- and UHDR-irradiated mice showed impaired novel object recognition. During fear learning, conventional- and UHDR-irradiated mice moved less during the inter-stimulus-interval (ISI) and UHDR-irradiated mice also moved less during the baseline period (prior to the first tone). In irradiated mice, reduced activity levels were also seen in the home cage; conventional- and UHDR-irradiated mice moved less during the light period and UHDR-irradiated mice moved less during the dark period. Following behavioral and cognitive testing, infiltrating immune cells in the hippocampus were analyzed by flow cytometry. The percent Ly6G+ CD45+ cells in the hippocampus was lower in conventional- and UHDR-irradiated than sham-irradiated mice, suggesting that neutrophils might be particularly sensitive to radiation. The percent of Ly6G+ CD45+ cells in the hippocampus was positively correlated with the time spent exploring the novel object in the object recognition test, suggesting that the reduced percent Ly6G+ CD45+ cells in the hippocampus might mediate some of the detrimental radiation-induced cognitive effects.

Cluster of differentiation 44 (CD44) promotes tumor progression through the recruitment of growth factors, and the acquisition of stemness, invasiveness, and drug resistance. CD44 has multiple isoforms including CD44 standard (CD44s) and CD44 variants (CD44v), which have common and unique functions in tumor development. Therefore, the elucidation of each CD44 isoform’s function in tumor is essential for the establishment of CD44-targeting tumor therapy. We have established various anti-CD44s and anti-CD44v monoclonal antibodies (mAbs) through immunization of CD44v3–10-overexpressed cells. In this study, we established C44Mab-6 (IgG1, kappa), which recognized the CD44 variant 3-encoded region (CD44v3) determined by enzyme-linked immunosorbent assay. C44Mab-6 reacted with CD44v3–10-overexpressed Chinese hamster ovary (CHO)-K1 cells (CHO/CD44v3–10) or some cancer cell lines (COLO205 and HSC-3) by flow cytometry. The apparent KD of C44Mab-6 for CHO/CD44v3–10, COLO205, and HSC-3 was 1.5 × 10−9 M, 6.3 × 10−9 M, and 1.9 × 10−9 M, respectively. C44Mab-6 could detect the CD44v3–10 in western blotting, and stained the formalin-fixed paraffin-embedded tumor sections in immunohistochemistry. These results indicate that C44Mab-6 is useful for detecting CD44v3 in various experiments, and expected for the application of tumor diagnosis and therapy.

Pancreatic cancer exhibits a poor prognosis due to the lack of early diagnostic biomarkers and the resistance to conventional chemotherapy. CD44 has been known as a cancer stem cell marker, and plays tumor promotion and drug resistance in various cancers. Especially, the splicing variants are overexpressed in many carcinomas, and play essential roles in the cancer stemness, invasiveness or metastasis, and resistance to treatments. Therefore, the understanding of each CD44 variant (CD44v) function and distribution in carcinomas is essential for the establishment of CD44-targeting tumor therapy. In this study, we immunized mice with CD44v3–10-overexpressed Chinese hamster ovary-K1 (CHO) cells, and established various anti-CD44 monoclonal antibodies (mAbs). One of the established clones (C44Mab-3; IgG1, kappa) recognized peptides of the variant 5-encoded region, indicating that C44Mab-3 is a specific mAb for CD44v5. Moreover, C44Mab-3 reacted with CHO/CD44v3–10 cells or pancreatic cancer cell lines (PK-1 and PK-8) by flow cytometry. The apparent KD of C44Mab-3 for CHO/CD44v3–10 and PK-1 was 7.1 × 10−10 M and 1.9 × 10−9 M, respectively. C44Mab-3 could detect the exogenous CD44v3–10 and endogenous CD44v5 in western blotting, and stained the formalin-fixed paraffin-embedded pancreatic cancer cells, but not normal pancreatic epithelial cells in immunohistochemistry. These results indicate that C44Mab-3 is useful for detecting CD44v5 in various applications, and expected for the application of pancreatic cancer diagnosis and therapy.

Leishmaniasis is a vector-borne parasitic infection caused by the bite of female Phlebotomine sandflies. World Health Organization (WHO) estimates 100,000 cases to be reported annually on a global scale, moreover, 13 million people were infected between 2010 and 2015 worldwide. Treatment of leishmaniasis by conventional synthetic compounds is met by challenges pertaining to adverse effects which call for the discovery of newer anti-leishmanial molecules. This study was performed to evaluate the effect and modes of action of a sesquiterpene alcoholic molecule Farnesol on Leishmania major, the causative agent of Zoonotic Cutaneous leishmaniasis. The cytotoxic effect of Farnesol against L. major promastigotes, amastigotes and macrophages was assessed by MTT test and counting. The IC50 on promastigotes by Farnesol on L. major was evaluated by flow cytometry. In the findings, Promastigotes were reduced at 167 µM/mL & the mean numbers of L. major amastigotes in macrophages were significantly decreased in exposure to Farnesol at 172 µM/ml. In addition, Farnesol demonstrated no cytotoxicity on macrophages as the IC50 value was 945 µM/ml; it induced significant apoptosis dose-dependent on L. major promastigotes. In silico protein-ligand binding analyses indicated the effect of Farnesol in perturbation of the ergosterol synthesis pathway of Leishmania with attributes suggesting inhibition of Lanosterol-α-demethylase, the terminal enzyme of ergosterol synthesis machinery. Findings from flow cytometry reveal the role of farnesol in apoptosis-induced killing in promastigotes. Farnesol was effective at very lower concentrations when compared to Paromomycin. Further studies are crucial to evaluate the therapeutic potentials of Farnesol alone and in combination with other conventional drugs using clinical settings.

CD44 has been known as a marker of tumor initiating cells, and plays pro-tumorigenic functions in many cancers. The splicing variants play critical roles in malignant progression of cancers by promoting the stemness, cancer cell invasion or metastasis, and resistance to chemo- and radiotherapy. To understand each CD44 variant (CD44v) function is essential to know the property of cancers and establishment of the therapy. However, the function of the variant 4-encoded region has not to be elucidated. Therefore, specific monoclonal antibodies (mAbs) against the variant 4 are indispensable for basic research, tumor diagnosis, and therapy. In this study, we established anti-CD44 variant 4 (CD44v4) monoclonal antibodies (mAbs) by immunizing mice with a peptide containing the variant 4-encoded region. We next performed flow cytometry, western blotting, and immunohistochemistry to characterized them. One of the established clones (C₄₄Mab-108; IgG₁, kappa) reacted with CD44v3-10-overexpressed Chinese hamster ovary-K1 cells (CHO/CD44v3-10). The K_D of C₄₄Mab-108 for CHO/CD44 v3-10 was 3.4 × 10⁻⁷ M. In western blot analysis, C₄₄Mab-108 detected CD44v3-10 in the lysate of CHO/CD44v3-10 cells. Furthermore, C₄₄Mab-108 stained formalin-fixed paraffin-embedded oral squamous carcinoma tissues in immunohistochemistry. These results indicated that C₄₄Mab-108 is useful to detect CD44v4 in various applications.

Background: Current strategies in circulating tumor cell (CTC) isolation in pancreatic cancer heavily rely on the EpCAM and cytokeratin cell status. EpCAM is generally not considered as a good marker given its transitory change during Epithelial to Mesenchymal Transition (EMT) or reverse EMT. There is a need to identify other surface markers to capture the complete repertoire of PDAC CTCs. The primary objective of the study is to characterize alternate surface bi-omarkers to EpCAM on CTCs that express low or negligible levels of surface EpCAM in pancre-atic cancer patients. Methods: Flow cytometry and surface mass spectrometry were used to iden-tify proteins expressed on the surface of PDAC CTCs in culture. CTCs were grown under condi-tions of attachment and in co-culture with naïve neutrophils. Putative biomarkers were then validated in GEMMs and patient samples. Results: Surface proteomic profiling of CTCs identi-fied several novel protein biomarkers. ALCAM was identified as a novel robust marker in GEMM models and in patient samples. Conclusions: We identified several novel surface bi-omarkers on CTCs expressed under differing conditions of culture. ALCAM was validated and identified as a novel alternate surface marker on EpCAMlow CTCs.

Yam (Dioscorea spp.) is a monocotyledonous herbaceous vine plant grown in the tropics and subtropics. It is a multi-species plant with varied intra- and interspecific ploidy levels. Of the 600 species, 11 are cultivated staple supporting the livelihood of over 300 million people. The paucity of information on ploidy and the genomic constitution is a significant challenge to the crop’s genetic improvement through crossbreeding. The objective of this study was to investigate the ploidy levels of 236 accessions across six cultivated and two wild species using chromosome counting, flow cytometry and genotyping-based ploidy determination methods. Results obtained from chromosome counting and genotyping-based ploidy determination were in agreement. In majority of the accessions, chromosome counting and flow cytometry were congruent, allowing future rapid screening of ploidy levels using flow cytometry. Among cultivated accessions, 168 (71%) were diploid, 50 (21%) were triploid, and 12 (5%) were tetraploid. Two wild species included in the study were diploids. Resolution of ploidy level in yams offers opportunities for implementing successful breeding programmes through intra- and inter-specific hybridization.

Objective: To investigate the effects and mechanisms of different concentrations of CCCP on mitophagy in human peripheral blood regulatory T cells. Methods: Tregs were isolated, identified and then grouped, treating with CCCP at a concentration of 2.5 μM, 5 μM, 10 μM, 20 μM and 40 μM for 24h in an incubator. Flow cytometry detected the reactive oxygen species (ROS), mitochondrial membrane potential (MMP), mitochondrial quality, and fluorescence microscopy observed the co-localization of mitochondria and lysosomes in each group. Results: The purity of CD4⁺CD25⁺Tregs was (93.36 ± 1.87) %. With the increase of CCCP concentration, the level of ROS gradually increased, while the MMP decreased gradually. About the mitochondria and lysosome fusion, the fluorescence intensity of orange (yellow) was the highest when the concentration of CCCP was in the range of 5-10 μM while decreased with the CCCP concentration continually increasing. The mitochondrial quality decreased with the increase of CCCP concentration. However, there was no significant difference between groups C, D and E. The mitochondrial quality of groups F and G were significantly lower than that of group E. Conclusions: With the concentration of CCCP gradually increased, the level of ROS in Treg cells increased, and MMP decreased, which promoted the mitophagy, mitochondrial quality maintains homeostasis; When ROS accumulated, and MMP decreased significantly, the mitophagy was inhibited, and the mitochondrial quality was significantly decreased.

One of the G protein-coupled receptors, C-C chemokine receptor 5 (CCR5), is an important regulator for the activation of T and B lymphocytes, dendritic cells, natural killer cells, and macrophages. Upon binding to the ligands, CCR5 activates downstream signaling, which plays critical roles in the innate and adaptive immune response through the promotion of lymphocyte migration and the secretion of pro-inflammatory cytokines. Anti-CCR5 monoclonal antibodies (mAbs) have been developed and evaluated in clinical trials for tumors and inflammatory diseases. In this study, we developed novel mAbs for mouse CCR5 (mCCR5) using the N-terminal peptide immunization. Among the established anti-mCCR5 mAbs, C₅Mab-4 (rat IgG_2a, kappa) and C₅Mab-8 (rat IgG₁, kappa), recognized mCCR5-overexpressed Chinese hamster ovary-K1 (CHO/mCCR5) and an endogenously mCCR5-expressing cell line (L1210) by flow cytometry. The dissociation constant (K_D) values of C₅Mab-4 and C₅Mab-8 for CHO/mCCR5 were determined as 3.5 × 10⁻⁸ M and 7.3 × 10⁻⁹ M, respectively. Furthermore, both C₅Mab-4 and C₅Mab-8 could detect mCCR5 by western blotting. These results indicated that C₅Mab-4 and C₅Mab-8 are useful for detecting mCCR5 by flow cytometry and western blotting and provide a possibility to obtain the proof of concept in preclinical studies.

Gastric cancer (GC) is the third leading cause of cancer-related deaths worldwide. GC with peritoneal metastasis exhibits a poor prognosis due to the lack of diagnostic biomarkers and effective therapy. A comprehensive analysis of malignant ascites identified the genomic alterations and significant amplifications of cancer driver genes, including CD44. CD44 and its splicing variants are overexpressed in tumors, and play crucial roles in the acquisition of invasiveness, stemness, and resistance to treatments. Therefore, the development of CD44-targeting monoclonal antibodies (mAbs) is important for GC diagnosis and therapy. In this study, we immunized mice with CD44v3–10-overexpressed PANC-1 cells and established several dozens of clones that produce anti-CD44v3–10 mAbs. One of the clones (C44Mab-94; IgG1, kappa) recognized the variant-8-encoded region and peptide, indicating that C44Mab-94 is a specific mAb for CD44v8. Furthermore, C44Mab-94 could recognize CHO/CD44v3–10 cells, oral squamous cell carcinoma cell line (HSC-3), or GC cell lines (MKN45 and NUGC-4) in flow cytometric analyses. C44Mab-94 could detect the exogenous CD44v3–10 and endogenous CD44v8 in western blotting and stained the formalin-fixed paraffin-embedded gastric cancer cells in immunohistochemistry. These results indicate that C44Mab-94 is useful for detecting CD44v8 in various applications and is expected to be useful for the application of GC diagnosis and therapy.

Cluster of differentiation 44 (CD44) has been investigated as a cancer stem cell (CSC) marker, and plays critical roles in tumor malignant progression. The splicing variants are overexpressed in many carcinomas, especially squamous cell carcinomas, and play critical roles in the promotion of tumor metastasis, the acquisition of CSC properties, and resistance to treatments. Therefore, each CD44 variant (CD44v) function and distribution in carcinomas should be clarified for the establishment of novel tumor diagnosis and therapy. In this study, we immunized mice with a CD44 variant (CD44v3−10) ectodomain and established various anti-CD44 monoclonal antibodies (mAbs). One of the established clones (C44Mab-34; IgG1, kappa) recognized a peptide which covers both variant 7 and 8-encoded region, indicating that C44Mab-34 is a specific mAb for CD44v7/8. Moreover, C44Mab-34 reacted with CD44v3–10-overexpressed Chinese hamster ovary-K1 (CHO) cells or oral squamous cell carcinoma (OSCC) cell line (HSC-3) by flow cytometry. The apparent KD of C44Mab-34 for CHO/CD44v3–10 and HSC-3 was 1.4 × 10−9 M and 3.2 × 10−9 M, respectively. C44Mab-34 could detect CD44v3–10 in western blotting, and stained the formalin-fixed paraffin-embedded OSCC in immunohistochemistry. These results indicate that C44Mab-34 is useful for detecting CD44v7/8 in various applications, and expected for the application of OSCC diagnosis and therapy.

The CC chemokine receptor 3 (CCR3) is a receptor for CC chemokines, including CCL5/RANTES, CCL7/MCP-3, and CCL11/eotaxin. CCR3 is expressed on the surface of eosinophils, basophils, a subset of Th2 lymphocytes, mast cells, and airway epithelial cells. CCR3 and its ligands are involved in airway hyperresponsiveness in allergic asthma, ocular allergy, and cancers. Therefore, CCR3 is an attractive target for those therapies. Previously, anti-mouse CCR3 (mCCR3) monoclonal antibodies (mAbs), C3Mab-3 (rat IgG2a, kappa), and C3Mab-4 (rat IgG2a, kappa) were developed using the Cell-Based Immunization and Screening (CBIS) method. In this study, the binding epitope of these mAbs was investigated using flow cytometry. The CCR3 extracellular domain-substituted mutant analysis showed that C3Mab-3, C3Mab-4, and a commercially available mAb (J073E5) recognized the N-terminal region (amino acids 1–38) of mCCR3. Next, the alanine scanning was conducted in the N-terminal region. The results revealed that Ala2, Phe3, Asn4, and Thr5 of mCCR3 are involved in C3Mab-3 binding, whereas Ala2, Phe3, and Thr5 are essential to C3Mab-4 binding, and Ala2 and Phe3 are crucial to J073E5 binding. These results reveal the involvement of the N-terminus of mCCR3 in the recognition of C3Mab-3, C3Mab-4, and J073E5.

C-C motif chemokine receptor-8 (CCR8) belongs to class A of G protein-coupled receptors (GPCRs). CCR8 interacts with the specific chemokine ligand CCL1/I-309 in humans, which is produced by various cells, including tumor-associated macrophages and regulatory T cells (Treg). CCR8 is highly expressed on Treg and T-helper 2 (T_H2) cells recruited to the inflammation site and is implicated in allergy, asthma, and cancer progression. The CCR8⁺Treg has been suggested to be an important regulator in the immunosuppressive tumor microenvironment (TME); therefore, it has been desired to develop sensitive monoclonal antibodies (mAbs) for CCR8. This study developed a specific mAb for human CCR8 (hCCR8), which is useful for flow cytometry by employing the Cell-Based Immunization and Screening (CBIS) method. The established anti-hCCR8 mAb, C₈Mab-21, (mouse IgM, kappa) reacted with hCCR8-overexpressed Chinese hamster ovary-K1 (CHO/hCCR8) cells, TALL-1 (acute T lymphoblastic leukemia), CCRF-HSB2 (human T-lymphoblastic leukemia), and natural killer cells, which express endogenous hCCR8 by flow cytometry. Furthermore, C₈Mab-21 demonstrated a moderate binding affinity for CHO/hCCR8 and TALL-1 with a dissociation constant of 6.5×10^-8 M and 2.0×10^-8 M, respectively. C₈Mab-21, which was established by the CBIS method, could be a useful tool for analyzing the hCCR8-related biological response using flow cytometry.

Diffuse large B-cell lymphoma (DLBCL) requires a complete staging at diagnosis that may have prognostic and therapeutic implications. The role of bone marrow (BM) biopsy (BMB) is contro-versial in the era of nuclear-imaging techniques. We performed a comparative review of 25 studies focused on BM evaluation at DLBCL diagnosis, including at least two of the following techniques: BMB, flow cytometry, and positron emission tomography (PET-FDG). The report about BM involvement (BMi), diagnostic accuracy and prognostic significance was collected and compared among techniques. A concordance analysis between BMB, FCM and PET was also per-formed, and we deeply evaluated the implications of the different types of BMi: concordant by LBCL or discordant by low-grade B-cell lymphoma for both BMB and FCM, and focal or diffuse uptake pattern for PET. As main conclusion, BMB, FCM and PET are complementary tools which provide different and clinically relevant information in the assessment of BMi in newly DLBCL.

In recent times, a great interest has been motivated in plant-derived compounds known as phytochemicals. The pentacyclic oleanane-, ursane-, lupane-type triterpenes are phytochemicals that exert significant activities against diseases like cancer. Lung cancer is the leading cause of cancer-related death worldwide. Although chemotherapy is the treatment of choice for lung cancer, its effectiveness is hampered by the dose‐limiting toxic effects and chemoresistance. Herein, we investigated the structure-activity relationship of six pentacyclic triterpenes: oleanolic acid (OleA), ursolic acid (UrA), asiatic acid (AsA), betulinic acid (BeA), betulin (Betu), lupeol (Lupe), on NSCLC A549 cells. From our studies, we confirmed that all these triterpenes showed cytotoxicity in the mid µM range, where BeA showed the strongest effect. Then, we determined that most of these triterpenes induced S-phase and G2/M phase cycle arrest, reactive oxygen species, mitochondrial depolarization, and caspase 3 activation. For chemoresistance markers, we elucidated that most triterpenes downregulated STAT3 and PDL1 genes. In contrast, ursane-type UrA and AsA also induced DNA damage, reactive nitrogen species and autophagy. These results showed that even slight structural changes in these triterpenes can influence the cellular response and mechanistic pathways. This study opens promising perspectives for further research on the pharmaceutical role of phytochemical triterpenoids.

A very important role in determining the quality of water is the assessment of its microbiological quality. Water quality control, which could pose a direct threat to human health and life, is performed in the case of water produced at the water treatment plants, tap water or water in swimming pools. However, these traditional methods used to assess its quality are laborious and time consuming. In emergency and incidental situations, in the era of terrorist threats, the need for quick, reliable and reproducible microbiological determinations seems to be essential. In this study, an attempt was made to compare various methods of assessing the microbiological quality of water. The assessment was carried out for waters with different microbiological characteristics: surface waters, rainwater, groundwater, and water supply. The evaluation was carried out using traditional culture methods and high-speed methods: flow cytometry and luminometry. The analysis of microbiological parameters were the basis for the statistical analysis. The conducted microbiological analysis of various types of water, along with their statistical evaluation, showed different dependencies for each of the analysed waters.

Pertussis is a vaccine-preventable disease caused by the bacterium Bordetella pertussis. Over the past years, the incidence and mortality of pertussis increased significantly. A possible cause is the switch from whole cell to acellular pertussis vaccines, although other factors may also contribute. To develop future vaccines and improve current vaccination strategies, it is critical to understand factors influencing the generation of immunological memory. We applied high-dimensional flow cytometry to investigate changes in B cells in individuals of different ages and distinct priming backgrounds upon administration of an acellular pertussis booster vaccine. These findings were correlated to vaccine-specific plasma cells and serum Ig levels. Expansion and maturation of plasma cells 7 days post-vaccination was the most prominent cellular change in all age groups, and was most pronounced for more mature IgG1+ plasma cells. Cellular responses were stronger in individuals primed with whole cell vaccine than in individuals primed with acellular vaccine. Moreover, IgG1+ plasma cell expansion weakly correlated with Prn- and PT- specific serum IgG levels. Our study points at plasma cells as a potential early cellular marker of an immune response and contributes to understanding differences in immune responses between age groups and priming backgrounds.

The growing production and application of carbon-based nanomaterials (CNMs) represent possible risks for aquatic systems. However, the variety of CNMs with different physical and chemical properties, and different morphology complicated the understanding of their potential toxicity. This paper aims to evaluate and compare the toxic impact of four most common CNMs, namely multiwalled carbon nanotubes (CNTs), fullerene (C60), graphene (Gr), and graphene oxide (GrO) in marine microalgae Porphyridium purpureum. The microalgae cells were exposed to the CNMs for 96 h and measured by flow cytometry. Based on the obtained results, we determined no observed effect level (NOEL), calculated EC10 and EC50 concentrations for growth rate inhibition, esterase activity, membrane potential, and reactive oxygen species (ROS) generation changes for each tested CNMs. According to the sensitivity (growth rate inhibition) of P. purpureum, the used CNMs can be listed in following order: CNTs &gt; GrO &gt; Gr &gt; C60. The toxicity of CNTs was significantly higher than the toxic effect of the other used CNMs and only this sample caused increase of ROS generation in microalgae cells. This effect caused by trace metal residuals in CNTs and high affinity between particles and microalgae associated with the presence of exopolysaccharide coverage on P. purpureum cells.

Beta-glucan-stimulated mammalian macrophages show an increased responsiveness to secondary stimulation in a nonspecific manner. This phenomenon is known as trained innate immunity. Our study aimed to explore training of primary chicken monocytes. We hypothesized that primary chicken monocytes, similar to their mammalian counterparts, can be trained with β-glucan resulting in increased responses of these cells to a secondary stimulus. Primary blood monocytes of white leghorn chickens were primary stimulated with β-glucan microparticulates (M-βG), LPS, recombinant chicken interleukin-4 (IL-4) or combinations of these components for 48 h. On day 6, the primary stimulated cells were secondary stimulated with LPS. Nitric oxide (NO) production levels were measured as an indicator of pro-inflammatory activity. In addition, the cells were analysed by flow cytometry to characterize the population of trained cells and to investigate the expression of surface markers associated with activation. After the secondary LPS stimulation, surface expression of CSF1R and the activation markers CD40 and MHC-II was higher on macrophages that were trained with a combination of M-βG and IL-4 compared to unstimulated cells. This increased expression was paralleled by enhanced NO production. In conclusion, this study showed that trained innate immunity can be induced in primary chicken monocytes with β-glucan, which is in line with previous experiments in mammalian species. Innate immune training may have potential to improve health and vaccination strategies within the poultry sector.

The C-C motif chemokine receptor 8 (CCR8) is highly and selectively expressed in regulatory T (Treg) cells and is associated with tumor progression. The massive accumulation of Treg cells into tumors suppresses the effector function of CD8⁺ cells against tumor cells. Therefore, selective depletion of Treg cells using anti-CCR8 monoclonal antibodies (mAbs) reinvigorates antitumor immune responses and improves responses to cancer immunotherapy. Previously, we developed an anti-mouse CCR8 (mCCR8) mAb, C₈Mab-2, using the Cell-Based Immunization and Screening (CBIS) method. In this study, the binding epitope of C₈Mab-2 was investigated using flow cytometry. The mCCR8 extracellular domain-substituted mutant analysis showed that C₈Mab-2 recognizes the N-terminal region (1–33 amino acids) of mCCR8. Next, 1×alanine (or glycine) scanning and 2×alanine (or glycine) scanning were conducted in the N-terminal region. The results revealed that the _17-DFFTAP_-22 sequence is important for the recognition by C₈Mab-2, and Thr20 is a central amino acid of the epitope. These results revealed the involvement of the N-terminus of mCCR8 in the recognition by C₈Mab-2.

Oxidoreductase (OXR) enzymes are in high demand for biocatalytic applications in food industry and cosmetics: glucose oxidase (GOx), cellobiose dehydrogenase (CBDH), bioremediation: horseradish peroxidase (HRP), laccase (LAC), and medicine for biosensors and miniature biofuel cells (GOx, CBDH, LAC, HRP). They can be used in a soluble form and/or within the yeast cell walls expressed as chimeras on the surface of yeast cells (YSD), such as P. pastoris and S. cerevisiae. However, most of the current studies suffer from either low yield for soluble enzyme expression or low enzyme activity when expressed as chimeric proteins using YSD. This is always the case in studies dealing with heterologous expression of oxidoreductase enzymes, since there is requirement not only for multiple OXR genes integrations into the yeast genome (super transformations), and codon optimization, but also very careful design of fermentation media composition, and fermentation conditions during expression due to need for adding transition metals (copper, iron), and metabolic precursors of FAD and heme. Therefore, scientists are still trying to find optimal formula using above mentioned approaches, and most recently also using protein engineering and directed evolution for additional increase in the yield of recombinant enzyme production. In this review article we will cover all the current state of the art technologies and most recent advances in the field that yielded high expression level for some of these enzymes in specially designed expression/fermentation systems. We will also tackle and discuss new possibilities for further increase in fermentation yield by the cutting-edge technologies such are directed evolution, protein and strain engineering, high-throughput screening methods based on in vitro compartmentalization, flow cytometry and microfluidics.

Malaria is rare in Europe, but it is a medical emergency and programs for its control should ensure access to early diagnosis and prompt treatment within 24-48 h of the symptom’s onset. The consistent number of imported malaria cases and the risk of reappearance of autochthonous cases stimulated laboratories in non-endemic countries to evaluate diagnostic methods/algorithms: microscopy remains the gold standard, but with limitations. RDTs have greatly expanded the ability to diagnose malaria for rapid results due to simplicity and low cost, but they lack sensitivity and specificity. PCR-based assays provide more relevant information but need well-trained technicians. According to WHO global Technical Strategy for Malaria 2016-2030, the future direction for a rapid diagnosis is the development of Point-Of-Care Testing, which would be helpful for prompt/accurate treatment and for preventing the diseases spread. Despite the limitations of current diagnostic methods, they play important roles in dealing with the global malaria situation, including decreasing its incidence. Recently evidence suggested that Artificial Intelligence could be utilized for assisting pathologists in malaria diagnosis. Future studies need to be conducted to see whether this system can be used for routine malaria diagnostic procedures.

The coronavirus disease caused by SARS-CoV-2 (known as COVID 19) is highly contagious and has spread rapidly over 200 countries over last three months. WHO suggested urgent escalation in testing, isolation and contact tracing, as the "backbone" of managing the pandemic. Globally, the detection of SARS-CoV-2 in patients are done by RT-PCR and blood antibody-based testing. In addition to the existing processes, flow-cytometry could be used as a high-throughput and efficient diagnostic method for detection of COVID 19. The suspected COVID 19 samples can be analyzed using ‘Indirect flow cytometry’, with the specific primary antibody and fluorescent tagged secondary antibodies. The fluorescence signal can distinguish the infected v/s non-infected samples. In the present article, we have summarized the applications of Flow virometry to study various viruses and have proposed possible application in the detection of COVID 19.

SARS-CoV-2 infection can lead to life-threatening clinical manifestations. Patients with cardiovascular disease (CVD) are at higher risk for severe courses of COVID-19. However, strategies to predict the course of SARS-CoV-2 infection in CVD patients at the time of hospital admission are still missing. Here, we investigated whether this prediction is achievable by prospectively analysing the blood immunophenotype of 94 participants, including 20 uninfected and 37 acutely SARS-CoV-2-infected CVD patients and 37 healthy donors, using a 36-colour spectral flow cytometry panel. Unsupervised data analysis revealed little differences between healthy donors and CVD patients, whereas the distribution of the cell populations changed dramatically in SARS-CoV-2-infected CVD patients. The latter had more mature NK cells, activated monocyte subsets, central memory CD4+ T cells, and plasmablasts but fewer dendritic cells, CD16+ monocytes, innate lymphoid cells, and CD8+ T cell subsets. Moreover, we identified an immune signature characterised by mucosal-associated invariant T (MAIT) cells, intermediate effector CD8+ T cells, and natural killer T (NKT) cells that is predictive for CVD patients with a severe course of COVID-19. Thus, intensified immunophenotype analyses can help identify patients at risk of severe COVID-19 at hospital admission, improving clinical outcomes through specific treatment.

Uncultured, unmodified, autologous, adipose-derived regenerative cells (UA-ADRCs) are a safe and effective treatment option for various musculoskeletal pathologies. However, it is unknown whether the composition of the final cell suspension systematically varies with the subject's individual age, sex, body mass index and ethnicity. UA-ADRCs were isolated from lipoaspirate from n=232 subjects undergoing elective lipoplasty using the Transpose RT system (InGeneron, Inc.; Houston, TX, USA). The UA-ADRCs were assessed for the number of nucleated cells, cell viability and the number of viable nucleated cells per gram of adipose tissue harvested. Cells from n=37 subjects were further characterized using four-channel flow cytometry. The present study shows, for the first time, that key characteristics of UA-ADRCs can be independent of the subject's age, sex, BMI and ethnicity. This result has important implications for the general applicability of UA-ADRCs in regeneration of musculoskeletal tissue. Future studies must determine whether in-dependence of key characteristics of UA-ADRCs of the subject's individual age, sex, BMI and eth-nicity only applies to the system used in the present study, or also to others of the more than 25 different experimental methods and commercially available systems used to isolate UA-ADRCs from lipoaspirate that have been described in the literature.

We report the first case of Daratumumab interference of allogeneic crossmatch tests repeatedly causing aberrant false-positive results, which inadvertently delayed transplant for a waitlisted renal patient with multiple myeloma. Daratumumab is an IgG1κ human monoclonal antibody commonly used to treat multiple myeloma, characterized by cancerous plasma cells and often leads to renal failure requiring kidney transplant, by depleting CD38-expressing plasma cells. In this case study, the patient had end-stage renal disease secondary to multiple myeloma and was continuously receiving Daratumumab infusions. The patient did not have any detectable antibodies to human leukocyte antigens but repeatedly had unexpected positive crossmatch by the flow cytometry-based method with 26 of the 27 potential deceased organ donors, implying donor-recipient immunological incompatibility. However, further review and analysis suggested that the positive crossmatches were likely false-positive as a result of interference from Daratumumab binding to donor cell surface CD38 as opposed to the presence of donor-specific antibodies. The observed intensity of the false-positive crossmatches was also highly variable, potentially due to donor- and/or cell-dependent expression of CD38. The variability of CD38 expression was, therefore, for the first time, characterized on the T and B cells isolated from various tissues and peripheral blood of 78 individuals. Overall, T cells were found to have a lower CD38 expression profile than the B cells, and no significant difference was observed between deceased and living individuals. Finally, we show that a simple cell treatment by dithiothreitol can effectively mitigate Daratumumab interference thus preserving the utility of pre-transplant crossmatch in multiple myeloma patients awaiting kidney transplant.

Mandevilla (Apocynaceae) is a greatly appreciated genus in the world ornamental market. In this study, we attempted to address the poor genetic knowledge and the huge taxonomic gaps existing in this genus by analyzing the germplasm of 55 accessions. After cytometrically determining the triploid genome size (1,512.64 Mb) of a reference sample (variety “Mandevilla 2001”), the plastidial genome (cpDNA, 0,18 Mb) and a draft of the nuclear genome (nuDNA, 207 Mb) were assembled. While cpDNA was effective in reconstructing the phylogenesis of the Apocynaceae family based on a DNA superbarcoding approach, the nuDNA assembly length was found to be only 41% of the haploid genome size (506 Mb, predicted based on the K-mer frequency distribution). Its annotation enabled the prediction of thousands of genes, of which 5,275 resulted in full CDS. Among them, we identified nine genes whose orthologs (in Catharanthus roseus) encode enzymes involved in the biosynthesis of monoterpene indole alkaloids (MIAs), including catharanthine, tabersonine and vincadifformine. The nuclear genome draft was also useful to develop a highly informative (mean PIC=0.62) set of 23 SSR markers that was validated on the Mandevilla collection. These results were integrated with cytometric measurements, nuclear ITS1 haplotyping and chloroplast DNA barcoding analyses to assess the origin, divergence and relationships existing among the 55 accessions object of the study. As expected, based on the scarce information available in the literature, the scenario was extremely intricate. A reasonable hypothesis is that most of the accessions represent interspecific hybrids sharing the same species as maternal parent (i.e., Mandevilla sanderi).

Artificial intelligence methods may help in unveliling information hidden in high-dimensional oncological data. Flow cytometry studies of haematological malignancies provide quantitative data with the potential to be used for the construction of response biomarkers. Many computational methods from the bioinformatics toolbox can be applied to these data but have not been exploited in their full potential in leukaemias, specifically for the case of childhood B-cell acute lymphoblastic leukemia. In this paper we analysed flow cytometry data obtained on diagnosis from 54 paediatric B-cell acute lymphoblastic leukemia patients from two local institutions. We constructed classifiers based on the Fisher’s Ratio to quantify differences in expression levels of immunophenotypical markers between patients with relapsing and non-relapsing disease. The distribution of the marker CD38 was found and validated to have a strong discriminating power between both patient cohorts, thus providing a classifier.

This work is related to the environmental toxicology risk assessment and evaluation of possible transformation of carbon-based nanomaterials (CNMs) after the contact with marine microalgae. The materials used in the study represent common and widely applied multiwalled carbon nanotubes (CNTs), fullerene (C60), graphene (Gr), and graphene oxide (GrO). The toxicity was evaluated as growth rate inhibition, esterase activity, membrane potential, and reactive oxygen species generation changes. The measurement was performed with flow cytometry after 3, 24, 96 h, and 7 days. Biotransformation of nanomaterials were evaluated after 7 days of microalgae cultivation with CNMs by FTIR and Raman spectroscopy. The calculated toxic level (EC50 in mg/L, 96 h) of ued CNMs reduced in the following order: CNTs (18.98) &gt; GrO (76.77) &gt; Gr (159.40) &gt; C60 (414.0). Oxidative stress and membrane depolarization was the main toxic action of CNTs and GrO. At the same time, Gr and C60 decreased the toxic action with time and had no negative impact on microalgae even at concentration of 125 mg/L. Moreover, C60 and Gr after seven days of the contact with microalgae cells obtained structural deformations.

Probiotics make up a large and growing segment of the commercial market of dietary supplements and are touted as offering a variety of human health benefits. Some of the purported positive impacts of probiotics include, but are not limited to, stabilization of the gut microbiota, prevention of gastrointestinal disorders and modulation of the host immune system. Current research suggests that the immunomodulatory effects of probiotics are strain specific and vary in mode of action. Here, we examined the immunomodulatory properties of Bacillus subtilis strain DE111 in a healthy human population. In a randomized, double blind, placebo-controlled four-week intervention, we examined peripheral blood mononuclear cells (PBMCs) at basal levels pre- and post-treatment as well as in response to stimulation with bacterial lipopolysaccharide (LPS). We observed an anti-inflammatory effect of B. subtilis, manifested as a decrease in immune cell populations within the basal state along with an increase in anti-inflammatory immune cells in response to LPS stimulation. Overall gastrointestinal health, microbiota, and circulating and fecal markers of inflammation and gut barrier function were largely unaffected by DE111 treatment. These data suggest that the novel probiotic B. subtilis DE111 may have clinical applications in modulating immune homeostasis via anti-inflammatory mechanisms.

Small cellular particles are released into the surroundings of cells and are proposed to play an important role in intercellular communication and consequently the responses of microbial com-munities to environmental stressors. We studied the connection between the small cellular parti-cles and the efficiency of three culture series of the microalge Phaeodactylum tricornutum and bac-teria (axenic microalgae, bacterial culture and co-culture of the two) in removing bisphenols from their growth medium. The microorganism growth rate was determined by flow cytometry, protein profiles were examined by protein gel electrophoresis, cultures and small cellular particle isolates were imaged by scanning electron microscopy, and bisphenols were analyzed using gas chroma-tography coupled with tandem mass spectrometry (GC-MS/MS). Higher growth rates of microal-gae were observed in the co-culture than in the axenic microalgal culture, while the presence of bisphenols neither influenced the morphology of the microalgal cells, protein profiles, nor the small cellular particle isolates. Biotic removal of bisphenols ranged from 0% to 71% and differed among the culture series in a compound-specific manner. However, it remains unclear which mechanisms influenced algal growth and bisphenol removal. Further research on the mechanisms of interspecies communication is needed to advance our understanding of microbial communities at the nano-level.

Background: A malfunction of the innate immune response in COVID-19 is associated with eosinopenia particularly in more severe cases. This study tested the hypothesis that this eosinopenia is COVID-19 specific and is associated with systemic activation of eosinophils.. Methods: Blood of 15 healthy controls and 75 adult patients with suspected COVID-19 at the ER were included before PCR testing and analyzed by point-of-care automated flow cytometry (CD10, CD11b, CD16 and CD62L) in the absence or presence of a formyl peptide (fNLF). 45 SARS-CoV-2 PCR positive patients were grouped based on disease severity. PCR negative patients with proven bacterial (n=20) or other viral (n=10) infections were used as disease controls. Eosinophils were identified with the use of the FlowSom algorithm. Results: Low blood eosinophil numbers (<100 cells/microL; p<0.005) were found both in patients with COVID-19 and with other infectious diseases albeit less pronounced. Two discrete eosinophil populations were identified in healthy controls both before and after activation with fNLF based on the expression of CD11b. Before activation, the CD11bbright population consisted of 5.4% (CI95% = 3.8, 13.4) of total eosinophils. After activation, this population of CD11bbright cells comprised nearly half the population (42.21%, CI95% = 35.9, 54.1). Eosinophils in COVID-19 had a similar percentage of CD11bbright cells before activation (7.6%, CI95% = 4.5, 13.6), but were clearly refractory to activation with fNLF as a much lower percentage of cells end up in the CD11bbright fraction after activation (23.7%, CI95% = 18.5, 27.6; p<0.001). Conclusion: Low eosinophil numbers in COVID-19 are associated with refractoriness in responsiveness to fNLF, which is likely caused by migration of fully functional cells to the tissue. Such homing might be beneficial as eosinophils have been implicated in viral killing.

Idiopathic lung fibrosis (IPF) is a progressive and fatal degenerative lung disease of unknown etiology. Although in its final stages it implicates in a reactive manner all lung cell types, the initial damage involves the alveolar epithelial compartment, in particular the alveolar epithelial type 2 cells (AEC2s). AEC2s serve dual progenitor and surfactant secreting functions, both of which are deeply impacted in IPF. Thus, we hypothesize that the size of the surfactant processing compartment, as measured by Lysotracker incorporation, allows the identification of different epithelial states in the IPF lung. Flow cytometry analysis of epithelial Lysotracker incorporation delineates two populations (Lysohigh and Lysolow) of AEC2s which behave in a compensatory manner during bleomycin injury and in the donor/IPF lung. Employing flow cytometry and transcriptomic analysis of cells isolated from donor and IPF lungs, we demonstrate that the Lysohigh population expresses all classical AEC2 markers and is drastically diminished in IPF. The Lysolow population, which is increased in proportion in IPF, co-expresses AEC2s and basal cell markers resembling the phenotype of the previously identified intermediate AEC2 population in the IPF lung. In that regard, we provide an in-depth flow-cytometry characterization of Lysotracker uptake, HTII-280, proSP-C, mature SP-B, NGFR, KRT5 and CD24 expression in human lung epithelial cells. Combining functional analysis with extra- and intra- cellular marker expression and transcriptomic analysis, we advance the current understanding of epithelial cell behavior and fate in lung fibrosis.

In addition to nucleic acids, various viral proteins are involved in the assembly of virion. If it is possible to create a biologically active agent that leads to the degradation of one of the key proteins for virus assembly and/or destroys the viral factory, then this agent will effectively cope with virus replication. One of these key proteins for the SARS-CoV-2 virus is the nucleocapsid protein (N-protein). As such a bioactive agent, we offer a modular nanotransporter (MNT) developed by us, which, in addition to an antibody mimetic to the N-protein, contains an amino acid sequence for the attraction of the Keap1 E3 ubiquitin ligase. This should lead to the subsequent degradation of the N-protein. We have shown that functional properties of modules within the MNT permit its internalization into target cells, endosome escape into the cytosol, and binding to the N-protein. Using flow cytometry and Western blot, it was demonstrated that significant degradation of N-protein is observed when A549 and A431 cells transformed with N-protein are incubated with the developed MNTs. The proposed MNTs open up a new approach for the treatment of viral diseases.

Background: Developing an efficient and standardized method to isolate and characterize adipose-derived stem cells (ASCs) from the stromal vascular fraction (SVF) of the adipose tissue for clinical application represents one of the major challenges in cell therapy and tissue engineering. Methods: In this study, we proposed an innovative, non-enzymatic protocol to collect clinically useful ASCs within freshly isolated SVF from adipose tissue by centrifugation of the infranatant portion of lipoaspirate and to determine the characteristic cytofluorimetric pattern, prior to in vitro culture. Results: The SVF yielded a mean of 73,32 \pm\ 10,89% cell viability evaluated with CALCEINA-FITC, i.e. cell-permeant dye. The ASCs were positive for PC7-labeled mAb anti-CD34 and negative for both PE-labeled mAb anti-CD31 and APC-labeled mAb anti-CD45. The frequency of ASCs estimated according to the panel of cell surface markers used was 51,06%\ \pm 5,26% versus the unstained ASCs subpopulation that was 0,74%\pm0,84% (P<0.0001). The ASCs events/\muL were 1602,13/\muL \pm 731,87/\muL. Conclusion: Our findings suggested that ASCs found in freshly isolated adipose SVF obtained by centrifugation of lipoaspirate can be immunophenotypically identified with a basic panel of cell surface markers. These findings aimed to provide standardization and contribute to reducing the inconsistency on reported cell surface antigens of ASC derived from the existing literature.

Pressure drop and flow pattern of oil-water flows were investigated in a 19 mm ID clear polyvinyl chloride pipe consisting of U-bend with radius of curvature of 100 mm. The range for oil and water superficial velocities tested were and respectively. Measurements were carried out under different flow conditions in a test section that consisted of four different parts: upstream of the bend, at the bend and at two redeveloping flow locations after the bend. The result indicated that the bend had limited influence on downstream flow patterns. However, the shear forces imposed by the bend caused some shift flow pattern transition and bubble characteristics in the redeveloping flow section after the bend relative to develop flow before the bend. Generally, pressure gradient at all the test sections increased with both oil fraction and water superficial velocity and there was a sharp change of pressure gradient profile during phase inversion. The transition point where phase inversion occurred was always within the range of . Pressure losses differed at the various test sections and the difference was strongly linked to the superficial velocity of the phases and the flow pattern. At high mixture velocity, pressure losses at the redeveloping section after the bend were higher than that at the bend and that for fully developed flows. At low mixture velocity, pressure losses at the bend are higher than in the straight sections. Pressure drop generally decreased with level of flow development downstream of the bend.

The flow properties of pellets or granules are crucial for further processing the drug dosage forms. Optimal compression or filling the multiparticulate dosage forms into capsules is influenced by forces between discrete particles, which could be partially characterized by flow properties. Several techniques were developed for the examination of flowability, including static and dynamic methods applying empirical studies and the up-to-date chaos theory; however newest methods seem only to be powerful with the supplement of empirical principles. Our experiments try to refine both the technique of analysis and the methods by finding new, alternative ways. Our approach to the flowability measurements was to set up a dynamic time-dependent model which combines empirical observations and the chaos theory on a geometrical basis, thus find new aspects regarding flow properties of pellets and granules that could be relevant for drug developers.

Juvenile myelomonocytic leukemia (JMML) is a deadly pediatric leukemia driven by RAS pathway mutations, of which >35% are gain-of-function in PTPN11. Although DNA hypermethylation portends severe clinical phenotypes, the landscape of histone modifications and chromatin profiles in JMML patient cells have not been explored. Using global mass cytometry, Epigenetic Time of Flight (EpiTOF), we analyzed hematopoietic stem and progenitor cells (HSPCs) from five JMML patients with PTPN11 mutations. These data revealed statistically significant changes in histone methylation, phosphorylation and acetylation marks that were unique to JMML HSPCs when compared with healthy controls. Consistent with these data, assay for transposase-accessible chromatin with sequencing (ATAC-seq) analysis revealed significant alterations of chromatin profiles at loci encoding post translational modification enzymes, strongly suggesting their mis-regulated expression. Collectively, this study reveals histone modification pathways as an additional epigenetic abnormality in JMML patient HSPCs, thereby uncovering a new family of potential druggable targets for the treatment of JMML.

This study will present a comprehensive review of the two-phase flow boiling heat transfer coefficient of hydrocarbons such propane (R-290), butane (R-600) and iso-butane (R-600a) and ethanol at various experimental conditions. Studying the multiphase flow heat transfer coefficient has a crucial importance for many heat transfer equipment to achieve higher efficiency for more compact design and cost reduction. One reason behind choosing hydrocarbons as refrigerants in this study is because hydrocarbons have zero ozone depletion potential (ODP=0) and insignificant direct global warming potential (GWP = 3). Moreover, thermodynamic and thermophysical characteristics of hydrocarbons qualify them to be a strong candidate for more heat transfer applications. Initially, by constructing a database for the working fluids from various experimental work available in the literature. The current data that this study has collected for the flow boiling of spans wide ranges of parameters, such as: mass flux, heat flux, operating pressure, and saturation temperature, etc. Furthermore, by comparing the experimental multiphase heat transfer coefficient database with the anticipated values of each correlation, the prediction performance of 26 correlations found in the literature was assessed. This study leads to the selection of the best prediction method based on the minimum deviation of predicted results from the experimental database provided by calculated mean absolute error (MAE) from the assessed correlations. The findings of this study can also be useful in the development of more accurate correlation methods for these fluids and improve the prediction of their flow boiling characteristics.

The article presents a multiphase numerical computational fluid dynamics (CFD) for simulating hydrothermal liquefaction of sewage sludge in a continuous plug-flow reactor. The discrete particle method (DPM) was used to analyze the solid particle interaction in liquid-solid high shear flows to investigated coupling computational fluid dynamics (CFD) and discrete phase model (DPM) method. Increasing solid particles interaction were observed with increasing liquid velocity. The study examined the influence of parameters such as flow rate, temperature and residence time on the efficiency of bio-oil production. Furthermore, the trajectories of the solid particles are affected significantly by the ash flow and the solid particle size, thus the trajectories of particles resembles the streamlines. A parametric study is performed to investigate the effect of slurry flow rate, temperature and external heat transfer coefficient on the biocrude oil productions performance. This simulation data will be used in the future to design and scale up a large-scale HTL reactor.

Optical micro-angiography (OMAG) is a new method of detecting flow rate and widely used for in vivo imaging. Although OMAG can distinguish between flowing and stationary parts, it cannot obtain accurate flow rate information. This study proposed a range formula for OMAG and the ultrahigh-sensitivity OMAG (UHS-OMAG) method to quantify the measurement range of an entire system. The parameters of the angle between beam scanning and flow directions, the angular velocity of the galvanometer, and the offset of incident light were introduced, and a formula for calculating the range was derived. Experiments were conducted to measure fine and ultra-fine flow rates by using OMAG and UHS-OMAG methods. The minimum measured flow rate was approximately 30 μm/s, and the maximum measured flow rate was approximately 8 mm/s. Experimental results are in good agreement with the preset results.

Oxidative stress has been implicated in cellular senescence and aging, as well as in the onset and progression of many diverse genetic and acquired diseases and conditions. The search for biomarkers of oxidative stress has become relevant to many biomedical fields using many different methods and approaches. Fluorescence methodology is advantageous because of its simplicity and high sensitivity. Fluorescent probes are frequently used to investigate the role of reactive oxygen species in oxidative stress in experimental or clinical settings, often involving the use of flow cytometry and related single-cell based technologies. To assess the specific role of reactive oxygen species in oxidative stress studies by cytometric methodologies, it is essential to detect and characterize these species accurately. However, the detection and quantitation of individual intracellular ROS is still a challenge, but different and complementary strategies may focus on other endpoints of oxidative stress. In this review we present and discuss briefly the limitations and perspectives of such approaches. Based on our results, we also provide recommendations for proper design of cytometric studies of oxidative stress in order to prevent or minimize experimental errors..

Mechanical ventilation systems, which are used for breathing support when a person is not able to do it by their own, requires a device for measuring the air flow to the patient in order to monitoring and a assure the magnitude establish by a medical staff. Flow sensors are the conventional devices used for the air flow measuring; however, there were not available in Peru, because of the international demand during COVID-19 pandemic. In this sense, a novel air flow sensor based on orifice plate and an intelligent transducer stage were developed as an integrated design. Advanced methodologies in simulations and experiments using specially designed equipment for this application were carried out. The obtained data was used for a mathematical characterization and dimensions validation of the integrated design. The device was tested in its real working conditions, it was implemented in a breathing circuit connected to a low-cost mechanical ventilation system based on cams. Results indicate that the designed air flow sensor/transducer is a low-cost complete medical device for mechanical ventilators able to provide satisfactorily all the ventilation parameters air flow, pressure and volume over time by measuring the air flow and calculating the others. Furthermore, this device provides directly a filtered equivalent electrical signal for a display or a computer.

Objectives: The purposes of this study were to 1) analyze the disruption of intracardiac flow by flow components and kinetic energy (KE) parameters in single ventricle physiology patients and compare with controls; 2) investigate left ventricular diastolic dysfunction in terms of 4D flow parameters in repaired Fontan (rFontan) group; 3) compare intracardiac flow parameters in morphologic LV and RV rFontan patient groups. Methods: Twenty-five rFontan patients (age: 10±3, M/F: 15/10) and fourteen controls (age: 10±2, M/F: 8/6) were prospectively recruited. All underwent cine and 4D flow cardiovascular magnetic resonance (CMR) on 3.0T scanner. Cardiac function and inter-ventricular mechanical dyssynchrony were analyzed using cine images. From 4D flow CMR, ventricular flow components were assessed: direct flow, retained inflow, delayed ejection flow and residual volume. Global and regional blood flow KE parameters, normalized to end-diastolic volume (EDV) were analyzed for functional single ventricle (FSV). Left ventricular (LV) diastolic dysfunction was assessed on echocardiography guidelines. Results: In comparison of rFontan vs. controls, median FSV residual volume (28% vs. 23%, P= 0.034) were higher in rFontan; median FSV direct flow (32% vs. 40%, P=0.005) and delayed ejection flow (17% vs. 24%, P=0.024) were lower in rFontan. FSV KEiEDV parameters were all lower in rFontan (all P<0.05). No significant differences were observed for flow components and KE parameters between patients with and without inter-ventricular dyssynchrony. FSV direct flow (AUC=0.76, Sensitivity=86%, Specificity=70%) was an independent predictor of LV diastolic dysfunction. Residual volume and E-wave KEiEDV were significantly different between morphologic RV and morphologic LV patient group. Conclusions: The changed flow pattern and decreased KE may be hemodynamic disturbances and impaired ventricular filling in rFontan patients. Reduced direct flow is associated with LV diastolic dysfunction. Morphologic right ventricular subgroup is worse than morphologic left ventricular group in terms of the intracardiac hemodynamics.

This study focuses on traffic condition analysis, especially in under developing country India and makes more visible of traffic flow by using traffic flow theory in order to understand real traffic condition. India is one of rapid economic growing countries and large market with second largest population 1.3 billion in 2018. On the other hand, there are social issues such as environment air pollution and global warming by traffic CO2 emission of transportation. This kind of condition is not only in India, but in other South East Asia and Africa in future. From recent more than one-month traffic observation in a typical major city Ahmedabad in Gujarat state, which has about 8 million population and industrialized city. In terms of traffic data collection, 14 CCTV cameras are used in the city. Based on multiple traffic cameras monitoring, author found the unique traffic flow characteristics and compares traffic flow theory. In this study, it is described what is key parameters to show real traffic congestion condition and how these congestion occurs.

The aim of this study is to model hydrodynamic processes of the Istanbul Strait with its stratified flow characteristic and calibrate the most important parameters using local and global search algorithms. For that two open boundary conditions are defined, which are in the North and South part of the Strait. Observed bathymetric, hydrographic, meteorological and water level data are used to set up the Delft3D-FLOW model. First, the sensitivities of model parameters on the numerical model outputs are assessed using PEST toolbox. Then, the model is calibrated based on the objective functions focusing on the flowrates of upper and lower layers. The salinity and temperature profiles of the Strait are only used for model validation. The results show that the calibrated model outputs of Istanbul Strait are reliable and consistent with the in-situ measurements. The sensitivity analysis reveals that the Spatial Low-Pass Filter Coefficient, Horizontal Eddy Viscosity, Prandtl-Schmidt Number, Slope in log-log Spectrum and Manning Roughness Coefficient are most sensitive parameters affecting flowrate performance of the model. The agreement between observed salinity profiles and simulated model outputs is promising whereas the match between observed and simulated temperature profiles is weak showing that the model can be improved particularly for simulating the mixing layer.

Hospital bed shortage is a worldwide concern. Beds unavailability has caused elective surgery cancellations, at our hospital peaking in spring 2016 at over 50%. This is often due to difficult patient step-down from intensive care (ICU) and high-dependency units (HDU). In our general/digestive surgery service admitting approximately 1000 patients yearly, ward rounds were run on a consultant firm basis. We report a quality improvement (ISRCTN13976096) introducing in our service a structured daily multi-disciplinary board round framework (SAFER Surgery R2G) adapted from the “SAFER patient flow bundle” and the "Red to Green days” approaches to enhance flow, comparing the previous year to 2017, when our framework was applied for 12 months.We used a Plan-Do-Study-Act (PDSA) methodology. Our intervention consisted in 1) systematic communication of the key care plan after the afternoon ward rounds to the nurse in charge; 2) 30’ 10AM Monday-to-Friday multidisciplinary board rounds, attended daily by the senior-team and weekly by hospital and site managers, revising the key care plan aiming at safe, early discharges, assessing the appropriateness of each inpatient day and tackling any cause of delay. We measured patient flow by average length of stay (LOS), ICU/HDU step-downs and operation cancellations count, monitoring safety through early 30-day readmissions. Compliance was assessed by board round attendance and staff satisfaction rate surveys.After 12 months (PDSA 2, N=1032), LOS significantly decreased from 7.2(±8.9) to 6.3(±7.4) days (p=0.003); ICU/HDU bed step-down flow increased by 9.3% from 345 to 375 (p=0.197), surgery cancellations dropped from 38 to 15 (p=0.100). 30-day re-admissions increased from 0.9% (N=9) to 1.3% (N=14)(p=0.390). Average cross-specialty attendance was 80%. Satisfaction rates were >75%, regarding enhanced teamwork and faster decisions.The SAFER Surgery R2G framework has shown an increase in patient flow in the context of an enhanced multidisciplinary approach, requiring senior staff commitment to remain sustainable.

Plant cells' ability to withstand abiotic stress is strongly linked to modifications in their mechanical characteristics. Nevertheless, the lack of a workable method for consistently tracking plant cell mechanical properties severely restricts our comprehension of the mechanical alterations in plant cells under stress. With Polyethylene Glycol (PEG6000), we created a drought-like environment, and we used the Double Resonator Piezoelectric Cytometry (DRPC) method to dynamically and non-invasively track changes in the stress (ΔS) created and viscoelasticity (storage modulus G' and loss modulus G") of protoplasts and suspension cells of rice during drought stress. The findings demonstrate that, rice suspension cells and protoplasts react mechanically differently to 5%–15% PEG6000 stress, implying distinct resistance mechanisms. However, neither of them can withstand 25% PEG6000 stress, they respond mechanically similarly to 25% PEG6000 stress. The results of DRPC are further corroborated by the morphological alterations of rice cells and protoplasts observed under an optical microscope. To sum up, the DRPC technique functions as a precise cellular mechanical sensor and offers novel research tools for the evaluation of plant cell adversity and differentiating between the mechanical reactions of cells and protoplasts under abiotic stress.

This study aimed to enhance a micro hydroelectric generator system driven by free-flow vortex and to compare efficiency of Propeller and Crossflow turbines. Series of turbines in each type were designed and tested at water-flowrate of 0.02 m3/s. The turbine housing has 1 meter in diameter and 0.5-meter height with 2 meters outlet drain at the bottom. The best efficiency extracted from Crossflow turbines with the same height (0.3 meter) but different in diameter (0.4, 0.5, 0.6, and 0.7 meter) and numbers of blade (12, 18, 24, 30, and 23) was from an 18 blades turbine at 23.01% of efficiency. The best efficiency extracted from Propeller turbines with 5 blades was from a 0.4-meter-high turbine with a diameter of 0.7 meter at 13.92% of efficiency. There were 12 Propeller turbines designed in this study. They were different in height (0.2, 0.3, and 0.4 meter) and, in each height, 0.4, 0.5, 0.6, and 0.7 of diameter was applied. The result revealed that Cross Flow turbine had more efficiency to the system than Propeller turbine (9.09%) at the water-flowrate of 0.02 m3/s

Design criteria for coastal defenses exposed to wave overtopping are usually assessed by mean overtopping discharges and maximum individual overtopping volumes. However, it is often difficult to give clear and precise limits of tolerable overtopping for all kind of layouts. A few studies analyzed the relationship between wave overtopping flows and hazard levels for people on sea dikes, confirming that one single value of admissible mean discharge or individual overtopping volume is not a sufficient indicator of the hazard, but detailed characterization of flow velocities and depths is required. This work presents the results of an experimental campaign aiming at characterizing the flow characteristics associated to maximum individual overtopping volumes for an urbanized stretch of a town along the Catalan coast, where a walking and bike path and a railway run along the coastline are exposed to significant overtopping events every stormy season. The work compares different safety criteria for pedestrian. Results prove that safety of pedestrian on a sea dike can be still guaranteed even for overtopping volumes larger than 1000 l/m. Pedestrian hazard is rather proved to be linked to the combination of overtopping flow velocity and flow depth.

The paper presents a modeling framework to analyze the effect of episodic rainfall supply on groundwater dynamics in the Ionian coastal plain multilayered aquifer. The focus is essentially on the short-term behavior of the shallowest layer, with a specific analysis on episodic rainfall events. In the studied aquifer, groundwater level responds sensitively to rainfall events, highlighting the presence of preferential recharge zones. The hydraulic head peak is a function of groundwater level antecedent to the rainfall event. A kinematic dispersion wave model was used to model infiltration processes via preferential pathways. A one-dimensional and non-linear particle based numerical model was developed. Particles with constant water volume travel according to celerity and hydraulic dispersion to simulate the infiltration rate wave through the vadose zone. The flow rate that reaches the water table represents the input function to determine groundwater level fluctuations along groundwater flow direction and according to the lithological features of the surficial levels of the multilayered aquifer, its storage capacity changes passing from unconfined to confined conditions. The model was validated with high time resolution time series of precipitation and groundwater level coming from Terra Montonata meteo-climatic and groundwater level monitoring station. The developed model represents a basis for evaluating and predicting groundwater discharge of the shallowest layers of the Ionian coastal multilayered aquifer under natural conditions including episodic rainfall.

Heart attacks and strokes are one of the leading causes of death in the world today, and heart attacks caused by clogged arteries that carry blood to the heart muscle are a significant part of these strokes. These are caused by the accumulation of fat particles in the walls of the arteries and the reduction of blood flow through it over a long process. The process of fat penetration in the underlying layers of the Artery wall has been the focus of many researchers, and various researches and Simulations have been done on it, in each of them, the effect of specific parameters has been considered. In the present study, the effect of blood flow rate on the flow pattern in a bifurcate artery with two ducts has been investigated using FLUENT software with Computation fluid dynamic Method. The effect of the angle between the two ducts of the Artery on the flow pattern has been investigated.

Using multi-color flow cytometry analysis, we studied the immunophenotypical differences between leukemic blasts/LSCs from patients with AML/MDS and hematopoietic stem and progenitor cells (HSPCs) from patients in complete remission (CR) following their successful treatment. The panel of markers included CD34, CD38, CD45RA, CD123 as representatives for a hierarchical hematopoietic stem and progenitor cell (HSPC) classification as well as programmed death ligand 1 (PD-L1). Rather than restricting the evaluation on a 2- or 3-dimensional analysis, we applied a t-distributed stochastic neighbor embedding (t-SNE) approach to get a deeper insight and segregation between leukemic and normal HPSCs. For that purpose, we created a t-SNE map, which resulted in the visualization of 27 cell clusters based on their similarity concerning the composition and intensity of antigen expression. Two of these clusters were “leukemia-related” containing a great proportion of CD34⁺/CD38^- hematopoietic stem cells (HSCs) or CD34⁺ cells with a strong coexpression of CD45RA/CD123, respectively. CD34⁺ cells within the latter cluster were also highly positive for PD-L1 reflecting their immunosuppressive capacity. Beyond this proof of principle study, the inclusion of additional markers will be helpful to refine the differentiation between normal HSPCs and leukemic cells, particularly in the context of minimal disease detection and antigen-targeted therapeutic interventions. Furthermore, we suggest a protocol for the assignment of new cell ensembles in quantitative terms, via a numerical value, the Pearson coefficient, based on a similarity comparison of the t-SNE pattern with a reference.

Debris flows have generated major disasters worldwide due to their great destructive capacity, which is associated with their high energy levels and short response times. To achieve an adequate risk management of these events, it is necessary to define as accurately as possible the different hazard levels to which the territory is exposed. This article develops a new methodology to estimate this hazard based on the hydrodynamic characteristics of the flow and the granulometry of the sediments that can be mobilized by the flow. The hydrodynamic characteristics of the flow are determined through a mathematical modeling that considers the rheology of non-Newtonian flows and the different volumes of sediments that could be transported during events corresponding to different return periods. The proposed methodology was implemented in the Jamundí River basin (Colombia). The results obtained indicate that in the upper part of this basin there is a low hazard level, while in the lower part of the basin approximately 15% of the affected territory has a medium hazard level and the remaining 85% has a low hazard level. The methodology developed is simple to implement but technically rigorous since it considers all relevant aspects in the generation of debris flows.

The two-phase flow of water and reactive gas in the channel of proton exchange membrane fuel cell (PEMFC) plays an important role in high-quality water management. In this study, the removal effect of 3D wave channel and 2D straight channel for liquid water was studied by volume of fluid method. The dynamic behavior of droplet emerge from the gas diffusion layer (GDL) into the channel under the influence of gas flow was investigated. The effects of droplet growth, deformation, detachment, force, pore size on water critical behavior and water content in the channel are discussed. The results show that the 3D wave channel is superior to the removal of liquid water than the 2D straight channel. The increase of velocity is conducive to the discharge liquid water. While infinite increase of velocity will cause more parasitic power loss. The larger pore size of GDL is not advantageous to the performance of PEMFC, but the appropriate increase of pore size is conducive to the discharge of liquid water. The basic knowledge obtained in this study will help to deepen the understanding of droplet dynamics in PEMFC gas channels, and help to optimize the design and working conditions of these channels.

Dams design and their operation cause strong environmental alteration and therefore a long-term debate is ongoing for the scale of these projects. At the same time, the concept of Environmental Flow Assessment (EFA) is a crucial element of modified ecosystems featuring large infrastructure such as dams and reservoirs for mitigating potential environmental degradation while they operate. Nowadays, integrated scientific frameworks are required to quantify the risks caused by large infrastructure. Through the use of stochastic analysis, it is possible to quantify these uncertainties, and present a solution that incorporates long-term persistence and environmental sustainability into a balanced reservoir simulation model. In this work, an attempt is made to determine a benchmark reservoir size incorporating hydrological and ecological criteria though stochastic analysis. The primary goal is to ensure the best possible conditions for the ecosystem, and then secondarily to allow a steady supply of water for other uses. Using a synthetic timeseries based on historical inputs, it is possible to determine and preserve essential statistical characteristics of a river’s streamflow, and use these to detect the optimal reservoir capacity that maximizes environmental and local water demand reliability.

Unsteady flow is the most common and complicated form of fluid motion in nature. This paper takes the beaches of key waterways in the middle and lower reaches of the Yangtze River as the research object, analyzes the water surface distribution and average flow velocity distribution near the beach, and analyzes the differences. The relationship between the flow pattern of the steep slope and the water flow pattern of the entire survey area and the steep slope of the beach and its nearby water flow structure. The water flow near the beach is divided into four major blocks: the swelling water area, the falling water area, the backwater area and the maximum flow rate area. The water flow structure characteristics and the cause of each block are analyzed.

Widely used in hydraulics, the Colebrook equation for flow friction relates implicitly to the input parameters; the Reynolds number, Re and the relative roughness of inner pipe surface, ε/D with the output unknown parameter; the flow friction factor, λ; λ=f(λ, Re, ε/D). In this paper, a few explicit approximations to the Colebrook equation; λ≈f(Re, ε/D), are generated using the ability of artificial intelligence to make inner patterns to connect input and output parameters in explicit way not knowing their nature or the physical law that connects them, but only knowing raw numbers, {Re, ε/D}→{λ}. The fact that the used genetic programming tool does not know the structure of the Colebrook equation which is based on computationally expensive logarithmic law, is used to obtain better structure of the approximations which is less demanding for calculation but also enough accurate. All generated approximations are with low computational cost because they contain a limited number of logarithmic forms used although for normalization of input parameters or for acceleration, but they are also sufficiently accurate. The relative error regarding the friction factor λ, in best case is up to 0.13% with only two logarithmic forms used. As the second logarithm can be accurately approximated by the Padé approximation, practically the same error is obtained also using only one logarithm.

The oscillating heat pipe (OHP) is a new member in the family of heat pipes, and it has great potential applications in energy conservation. However, the fluid flow and heat transfer in the OHP as well as the fundamental effects of inner diameter on them have not been fully understood, which are essential to the design and optimization of the OHP in real applications. Therefore, by combining the high-speed visualization method and infrared thermal imaging technique, the fluid flow and thermal performance in the OHPs with inner diameters of 1, 2 and 3 mm are presented and analyzed. The results indicate that three fluid flow motions, including small oscillation, bulk oscillation and circulation, coexist or, respectively, exist alone with the increasing heating load under different inner diameters, with three flow patterns occurring in the OHPs, viz. bubbly flow, slug flow and annular flow. These fluid flow motions are closely correlated with the heat and mass transfer performance in the OHPs, which can be reflected by the characteristics of infrared thermal images of condensers. The decrease in the inner diameter increases the frictional flow resistance and capillary instability while restricting the nucleate boiling in OHPs, which leads to a smaller proportion of bubbly flow, a larger proportion of short slug flow, a poorer thermal performance, and easier dry-out of working fluid. In addition, when compared with the 2 mm OHP, the increasing role of gravity induces the thermosyphon effect and weakens the 'bubble pumping' action, which results in a little smaller and bigger thermal resistances of 3 mm OHP under small and bulk oscillation of working fluid, respectively.

In this paper, we present the extension of the pressure-based solver designed for the simulation of compressible and/or incompressible two-phase flows of viscous fluids. The core of the numerical scheme is based on the hybrid Kurganov— Noele — Petrova/PIMPLE algorithm. The governing equations are discretized in the conservative form and solved for velocity and pressure, with the density evaluated by an equation of state. The acoustic-conservative interface discretization technique helps to prevent the unphysical instabilities on the interface. The solver was validated on various cases in wide range of Mach number, both for single-phase and two-phase flows. The numerical algorithm was implemented on the basis of the well-known open-source Computational Fluid Dynamics library OpenFOAM in the solver called interTwoPhaseCentralFoam. The source code and the pack of test cases are available on GitHub: https://github.com/unicfdlab/hybridCentralSolvers

The turbulent Poiseuille flow between two parallel plates is one of the simplest possible physical situations, and it has been studied intensively. In this paper, we propose a modified Prandtl-van Driest mixing length that satisfies both boundary conditions and wall damping effects. With our new formulations, we numerically solve the problem and, moreover, propose an approximate analytical solution of mean velocity. As applications of our solution, an approximate analytical friction coefficient of turbulent Poiseuille flow is proposed.

The problem of fluid dynamics can be greatly simplified if, for every point in space, the strain-rate tensor is diagonalized. This tensor is introduced into the Navier-Stokes equations via material law and divergence of the stress tensor. This article shows that local SO(3)xU(1) gauge fields can be used to locally diagonalize the diffusion components of the strain-rate tensor. The gauge fields resulting from the connection can be interpreted as convection components of the flow, they show properties of quasiparticles and can be interpreted as elementary vortices. Thus, the proposed approach not only offers new insights for the solution and situative simplification of the Navier-Stokes equations, it also uncovers hidden symmetries within the flow convection, allowing - depending on boundary conditions - further interpretation.

Aero-fuel centrifugal pumps are important power plants in aero-engines. Unlike most of the existing centrifugal pumps, a combination impeller is integrated with the pump to improve its performance. First, the critical geometrical parameters of combination impeller and volute are given. Then, the effects of combination impeller on flow characteristics inside the impeller and volute are clarified by comparing simulation results with that of the conventional impeller, where the effectiveness of selected numerical method is validated by an acceptable agreement between simulation and experiment. Finally, the experiment is performed to test the external performance of studied pump. A significant feature of this study is that the flow characteristics are significantly ameliorated by reducing the flow losses emerged in impeller inlet, impeller outlet and volute tongue. Correspondingly, the head and efficiency of combination impeller are higher with comparison to conventional impeller. Consequently, it is a promising approach in ameliorating flow field and improving external performance by applying a combination impeller to an aero-fuel centrifugal pump.

Deformation flows are flows incorporating shear, strain and rotational components. These flows are ubiquitous in the geophysical flows, such as the ocean and atmosphere. They appear near almost any salience, such as isolated coherent structures (vortices and jets), various fixed obstacles (submerged obstacles, continental boundaries). Fluid structures subject to such deformation flows may exhibit drastic changes in motion. In this review paper, we focus on the motion of a small number of coherent vortices embedded in deformation flows. Problems involving isolated one and two vortices are addressed. When considering a single-vortex problem, the main focus is on the evolution of the vortex boundary and its influence on the passive scalar motion. Two vortex problems are addressed with the use of point vortex models, and the resulting stirring patterns of neighbouring scalars are studied by a combination of numerical and analytical methods from the dynamical system theory. Many dynamical effects are reviewed with emphasis on the emergence of chaotic motion of the vortex phase trajectories and the scalars in their immediate vicinity.

Since its introduction almost 50 years ago, the concept of ‘flow’ has been descriptive of optimal experiences, also in relation to play. However, the explorative nature of play leads to some discrepancies between flow and the optimal experience of play. In this paper the differences between flow and play are explored, leading to proposing the state of ‘wonder’ (directed at exploration) as an alternative to ‘flow’. From this perspective, the study further explores how we may design toys that enable meaningful experiences of play, identifying opportunities in designing for toys as the enablers of immersed experiences of wonder.

There is a long standing ‘two times problem’ in that a satisfactory reconciliation between the time of physics and that of psychology has not been realized. A partial solution to the past/present/future phenomenon has been successfully given by the Hartle information gathering and processing system (IGUS) view. That model IGUS robot is enhanced here for the entire ‘two times problem’ to deal with not only the temporal experiences of the flow of time but also those of manifest time. A dualistic robot is proposed which has a veridical system of temporal experiences that are compatible with various spacetime cosmologies. It also has an illusory system of corresponding temporal experiences. This dualistic IGUS robot was made possible by discovering temporal experience within the brain that correspond to those of physics. The dualistic theory suggests that the veridical system, as a result of evolution, begets the illusory system to enhance behavioral adaptation. Thus, there is just one fundamental physical time which the brain does, indeed, possess and then enhances with illusory counterparts. Therefore, there should no longer be a need to reify illusory temporal experiences as modern spacetime cosmologies tend to do. Physical time already resides within human time.

In this study, the author proposes to evaluate the effect of sales growth, Receivable Turnover and operating cash flow on the liquidity of PT. Unilever Indonesia Plc. The research method used is descriptive method with a quantitative approach. In this statement, the population used in this study is the financial statement data from PT. Unilever Indonesia Plc. from 2010 to 2018, the technique of determining the sampling uses Purposive Sampling. This research data uses secondary data from PT. Unilever Indonesia Plc financial statements from 2010 to 2018. All data sources were obtained from the website of the Indonesia Stock Exchange at https://www.idx.co.id, the company's website and Google search. Our analysis reveals that sales growth and accounts receivable turnover from PT. Unilever Indonesia Plc. has no influence on the liquidity of PT. Unilever Indonesia Plc, while operating cash flow has an influence on the Liquidity of PT Unilever Indonesia Plc. This means the ups and downs of the value of sales and accounts receivable turnover of a company has no influence on the liquidity of PT. Unilever Indonesia Plc, while operating cash flow has increased or decreased has an influence on the liquidity of PT Unilever Indonesia Plc. The value of sales growth, accounts receivable turnover and operating cash flow can explain the liquidity of PT Unilever Indonesia Plc. by 78%, while 22% is explained by other factors which are not included in this study.

Rainfall is the main driver of hydrological processes in dryland environments and characterising the rainfall variability and processes of runoff generation are critical for understanding ecosystem function of catchments. Using remote sensing and in situ data sets, we assess the spatial and temporal variability of the rainfall, rainfall-runoff response, and effects of antecedent soil moisture and ground cover at different spatial scales on runoff coefficients in the Upper Burdekin catchment, northeast Australia, which is a major contributor of sediment and nutrients to the Great Barrier Reef. The high temporal and spatial variability of rainfall exerts significant controls on runoff generation processes. Rainfall amount and intensity are the primary runoff controls, and runoff coefficients for wet antecedent conditions were higher than for dry conditions. The majority of runoff occurred via surface runoff generation mechanisms, with subsurface runoff likely contributing little runoff due to the intense nature of rainfall events. At annual to seasonal temporal scales and for relatively large catchments, we could not detect a significant effect of ground cover on runoff. We conclude that in the range of moderate to large catchments (193 – 36,260 km²) runoff generation processes are sensitive to both antecedent soil moisture and ground cover. A higher runoff-ground cover correlation in drier months with sparse ground cover highlighted the critical role of cover at the onset of the wet season and how runoff generation is more sensitive to cover in drier months than in wetter months. The monthly water balance analysis indicates that runoff generation in wetter months (January and February) is partially influenced by saturation overland flow, most likely confined to saturated soils in riparian corridors, swales, and areas of shallow soil. By March and continuing through October, the soil ‘bucket’ progressively empties by evapotranspiration, and Hortonian overland flow becomes the dominant, if not exclusive, flow generation process. The results of this study can be used to better understand the rainfall-runoff relationships in dryland environments and subsequent exposure of coral reef ecosystems in Australia and elsewhere to terrestrial runoff.

Maxwell equations governing electromagnetic effects are being shown to be equivalent to the compressible inviscid Navier-Stokes equations applicable in fluid dynamics and representing conservation of mass and linear momentum. The latter applies subject to a generalized Beltrami condition to be satisfied by the magnetic field. This equivalence indicates that the compressible inviscid Navier-Stokes equations are Lorentz invariant as they derive directly from the Lorentz invariant Maxwell equations subject to the same Beltrami condition. In addition, the derivation results provide support for the claim that electromagnetic potentials have physical significance as demonstrated by Aharonov-Bohm effect, and are not only a convenient mathematical formulation.

Temporary streams are a key component of the hydrological cycle in arid and semi-arid regions, but their flow is highly variable and difficult to measure. In this paper, we present a novel approach that could be used to assess the flow of temporary streams and characterize their environmental status. Specifically, we apply the Image Velocimetry method to estimate surface velocity in temporary streams using unmanned aerial vehicles (UAVs) equipped with optical sensors (IV-UAV method). The IV-UAV method enables the easy, safe and quick estimation of the velocity on the water’s surface. This method was applied in different temporary streams in Lesvos Island, Greece. The produced results show that the IV-UAV can be implemented at low discharges, at temporary streams and at small streams. However, there were many occasions that measurements were unable due to various reasons such as dense vegetation or archaeological sites. Despite of this, the proposed methodology could be incorporated in optical protocols which are used to assess the environmental status of temporary streams of Mediterranean conditions. Finally, this would become a valuable tool for understanding the dynamics of these eco-systems and monitoring changes over time.

The discrete element method (DEM) coupled with computational dynamics (CFD) has been considered one of the most sensitive ways of studying fluidized beds. This paper presents gas-solid fluidized bed simulations by use of CFD-DEM. The two-phase interaction is calculated in a way of decoupling. Within the effective neighborhood of the particles, the kernel approximation method is used to calculate the local porosity, thereby calculating the drag force on the particles. At the grid scale, the momentum exchange coefficient is calculated using a two fluid model based method, thereby calculating the source term of the phase interaction in the fluid control equation. The simulated form, size and motion process of the big bubble are all consistent with experimental observations. The simulated solid volume fraction and relative pressure averaged in a horizontal plane at 45 mm above the bottom are in reasonable agreement with the experimental data. The fluctuation of the simulated bed height over time is similar to the experimental measurement results. The reproduction of these qualitative and quantitative results indicates that the proposed method effectively improves simulation performance and accuracy.

Active flow control is a promising technology for reducing noise, emissions, and power consumption in various applications. To better understand the performance of synthetic jet actuators, a computational model that couples structural mechanics with electrostatics, pressure acoustics, and fluid dynamics has been needed. The model presented here has been validated against experimental data and then used to investigate the fluid behavior inside and outside the synthetic jet actuator cavity, impacts of thermoviscous losses on capturing the acoustic response of the actuator, and viability of different modeling methods of diaphragms in computational simulations. The results capture the feedback from the fluid onto the diaphragm and highlight the need for careful acoustic modeling.

Material and methods. The trial was carried out over two years in Southern Italy. Two grapevine rootstocks, 110R and SO4, were compared to evaluate the water extraction ability from the soil and the effect on the yield and quality of the Cardinal grapevine table cultivar. Therefore, a new approach to plant water consumption based on sap flow was adopted. Results. The earlier and faster water refilling of the xylem in Cardinal onto 110R (C/110R) appears responsible for the earlier evolution of the phenological phases than Cardinal onto SO4 (C/SO4). The maximum length of the principal shoot was reached in Cardinal/110R compared to C/SO4, while a higher number of lateral shoots with lower internode has changed canopy architecture and light distribution in C/SO4. The 110R used more water compared to SO4; it was possible to quantify the real transpired flux of the plant per day: sap flow was 12.3 L.plant-1.d-1 and 11.7 L.plant-1d-1 in C/110R in the first and second year, respectively, while it was 14% lower in alternative graft combination. Conclusions. However, C/SO4 was able to sustain leaf water status and physiological mecha-nisms, although with lesser performance than C/110R, but that did not cause negative effects on production parameters.

With the wide application of flow sensor, their reliability under extreme conditions has been concerned in recent years. The reliability of a Micro Electro Mechanical Systems flow sensor under temperature (Ts) is researched in this paper. Firstly, the Step-stress accelerated degradation test is designed. Because this flow sensor consists flow sensor chip and signal processing system, the accelerated degradation testing is implemented, respectively. While the results show that the biggest drift is 3.15% for flow sensor chips with 150℃ conditions, and 32.91% for this flowmeter. By analysis, it could be found that the attenuation of the signal processing system is significant to the degeneration of this flowmeter. The minimum drift of the signal processing system accounts for 82.01% of this flowmeter. Secondly, using the Coffin-Manson model, the relationship between cycle-index and Ts is established. And the lifetime with different Ts is estimated by the Arrhenius model. In addition, the Weibull distribution is applied to evaluate the lifetime distribution. Finally, the reliability function of the Weibull distribution is demonstrated, and the survival rate within one year is 87.69% with 85℃ conditions. Thus, by the application of accelerated degradation testing, the acquired results are innovative and original. This research illustrates the reliability research, which provides a relational database for the application of this flow sensor.

Background: Poor patient flow can lead to adverse outcomes for patients and organisational inefficiency. Many hospitals have addressed suboptimal patient flow by increasing resourcing, such as bed stock and staffing, however, this is an unsustainable approach. In determining the nature of poor patient flow issues, it is important to collect data from healthcare professionals who manage patient flow on a daily basis. Doing so provides insights into the current state of patient flow management in its entirety, whilst also helping with the development of sustainable solutions.; Methods: Thirteen semi-structured interviews were conducted with clinicians who were directly involved with patient flow at a tertiary care centre in Tasmania, Australia. Results: Through a thematic analysis method, four major themes were developed: managing patient flow, communication for decisions, tools as enablers and barriers and increasing complexity. Conclusions: The findings of this study provide great insights into patient flow issues, with potential solutions identified to address them.

This paper presents the results of simulations of particle-laden air-solid jet flow in long straight tubes using CFD-DEM, along with an analysis of coarse-graining. Although previous studies have used CFD-DEM for similar flows, these have typically been in a dilute flow regime where uncoupled simulations are accurate. However, fully coupled simulations can introduce issues, necessitating validation studies to ensure that all coupling parameters are effectively used and that the physics is accurately represented. This paper validates the simulations against two different experimental studies, with fluid Reynolds numbers between 10,000 and 40,000 and Stokes numbers between 5.6 and 50. Interestingly, the profiles of mean particle velocity exhibit fewer discrepancies as the Stokes number increases but more discrepancies for the root mean square velocity compared to the experiments. Particle number flux is consistent with the experiments after the nozzle exit. Coarse-graining is also applied to the same simulations, achieving relatively accurate results. However, as expected, scaling of contact collision frequencies, forces, and stresses cannot be achieved, meaning that coarse-graining may be useful for comparing designs or operating parameters on an industrial scale but falls short when measuring the total energy dissipation of one experiment.

Gas embolism is a medical condition leading to the blockage of blood flow in the microvasculature by gas bubbles. While reported as rare, gas embolism often has devastating, fatal physiological consequences. Despite this acute importance, the genesis and evolution of air bubbles in blood vessels under different physiological conditions, such as blood viscosity and blood flow rate, is still understudied, largely because of difficult experimentation and in situ visualization. The objective of this work was to study the gas embolism phenomenon in vitro, using a microfluidic system that mimicked the architecture of microvasculature. The microfluidic systems comprised linear channels with two different air inlet types, namely, T- and Y-junctions with three different widths (20 µm, 40 µm, and 60 µm), and a 30 µm width honeycombed network with three bifurcation angles (30°, 60°, and 90°). Three synthetic liquids equivalent to 0%, 20%, and 46% hematocrit that mimicked the physiological blood viscosity and hematocrit concentrations were used. Our results show that: (i) 20 µm and 40 µm width channels had an elevated risk of gas embolism due to wide fluctuations in the total slug sizes; (ii) the resistance to the flow of air bubbles increased with the increase in the equivalent concentration of hematocrit; (iii) gas bubbles causing blockages and dampening of the flow velocity were frequently observed in 20 µm channels, and lastly (iv) increased risk of gas embolism was observed in the honeycomb architecture with 60° and 30° bifurcations. This work suggests that in vitro experimentation using microfluidic devices with microvascular tissue-like structures opens the possibility of studying this medical condition with high reproducibility and impacts the fact-based medical guidelines for preventing or mitigating iatrogenic occurrences.

This paper presents an analysis of linear viscous stress Favre-Averaged turbulence models for computational fluid dynamics (CFD) of fully turbulent round jets with a long straight tube geometry in the near field. Although similar work has been performed in the past with very relevant solutions, considerations were not given for the issues and limitations involved with coupling between an Eulerian and Lagrangian phase, such as in fully two-way coupled CFD-DEM. These issues include limitations on solution domain, mesh cell size, wall modelling, and momentum coupling between the two phases in relation to the particles size. Therefore, within these considerations, solutions are provided to the Navier-Stokes equations with various turbulence models using a three-dimensional wedge long straight tube geometry for fully developed turbulence flow. Simulations are performed with a Reynolds number of 15000 and 50000 using two different tube diameters. It is found that a modified k−ε turbulence model achieved the most agreeable results for both the velocity and turbulent flow fields between these two flow regimes, while a modified k−ω SST/BSL also provided suitable results.

Evidence supports an association between low magnesium (Mg) intake and coronary heart disease and between Mg intake and endothelial function. The aim of this study was to assess the effect of one week of Mg supplementation on endothelial function, assessed by flow mediated dilatation (FMD). Nineteen healthy men and women completed this cross-over pilot study in which participants were randomised to take an over-the-counter magnesium supplement for one week or to follow their usual diet. Weight, FMD and blood pressure (BP) were taken on completion of each intervention and 24hour urine collections and blood samples were taken to assess compliance. Baseline serum Mg was within normal range for all participants. Urinary Mg and urinary magnesium-creatinine ratio (Mg/Cr) significantly increased between interventions, (p = 0.03, p = 0.005 respectively). No significant differences in FMD or BP were found between the interventions. A significant negative correlation was seen between age and FMD (r = -0.496, p = 0.031). When adjusted for age, saturated fat was negatively associated with FMD (p = 0.045). One week of Mg supplementation did not improve FMD in a healthy population.

Evidence supports an association between low magnesium (Mg) intake and coronary heart disease and between Mg intake and endothelial function. The aim of this study was to assess the effect of one week of Mg supplementation on endothelial function, assessed by flow mediated dilatation (FMD). Nineteen healthy men and women completed this cross-over pilot study in which participants were randomised to take an over-the-counter magnesium supplement for one week or to follow their usual diet. Weight, FMD and blood pressure (BP) were taken on completion of each intervention and 24hour urine collections and blood samples were taken to assess compliance. Baseline serum Mg was within normal range for all participants. Urinary Mg and urinary magnesium-creatinine ratio (Mg/Cr) significantly increased between interventions, (p = 0.03, p = 0.005 respectively). No significant differences in FMD or BP were found between the interventions. A significant negative correlation was seen between age and FMD (r = -0.496, p = 0.031). When adjusted for age, saturated fat was negatively associated with FMD (p = 0.045). One week of Mg supplementation did not improve FMD in a healthy population.

Magnesium (Mg) deficiency may be a catalyst in the process of endothelial dysfunction, an early event in the pathogenesis of atherosclerosis. The aim was to determine the acute effect of an oral Mg supplement compared to control on endothelieal function assessed by flow-mediated-dilatation (FMD). Nineteen participants (39 years, body mass index (BMI) 22.9kg/m2) completed this randomised cross-over study. Blood pressure (BP) and FMD were measured and blood samples taken before participants drank 200ml water with or without an over the counter Mg supplement (450mg and 300mg for men and women). Measurements were repeated at 60 and 120 minutes. There was a statistically significant two-way interaction between treatment and time on serum Mg (p = .037). A difference of -0.085mm in FMD was observed 60 minutes post drink in the control group compared to baseline FMD, and no difference was observed in the supplement group compared to baseline. Despite the non-significant interaction between treatment and time on FMD, the difference seen in the control group and the lack of change in the supplement group at 60 minutes post-drink suggests that Mg may attenuate the reduction in FMD post-prandially.

89Zr-radiolabelled proteins functionalised with desferrioxamine B are a cornerstone of diagnostic positron emission tomography. In the clinical setting, 89Zr-labelled proteins are produced manually. Here, we explore the potential of using a microfluidic photochemical flow reactor to prepare 89Zr-radiolabelled proteins. The light-induced functionalisation and 89Zr-radiolabelling of human serum albumin ([89Zr]ZrDFO-PEG3-Et-azepin-HSA) was achieved by flow photochemistry with a decay-corrected radiochemical yield (RCY) of 31.2±1.3% (n = 3) and radiochemical purity &gt;90%. In comparison, a manual batch photoreactor synthesis produced the same radiotracer in a decay-corrected RCY of 59.6±3.6% (n = 3) with an equivalent RCP &gt;90%. The results indicate that photoradiolabelling in flow is a feasible platform for the automated production of protein-based 89Zr-radiotracers, but further refinement of the apparatus, and optimisation of the method is required before the flow process is competitive with manual reactions.

The objective of this work is to evaluate the impact of the innovative modifications made to the conventional effluent capture and discharge devices used in subsurface flow wetlands (SSFW). The main modifications developed consist on extending the influence of the capture and discharge device in such a way that the SSFW width and height are fully covered. This improved innovative device was applied and evaluated in two subsurface flow wetlands, one on a pilot scale and one on a real scale. To evaluate the impact of the innovative device with respect to conventional one in the operational functioning of subsurface flow wetlands, the elimination of chemical oxygen demand was measured and compared. The results show that for the innovative device, the COD removal was 10% higher than for the conventional device, confirming the validity and effectiveness of the modifications implemented in the effluent capture and discharge devices used in SSFW.

The hydrodynamic results obtained from the permeability of porous materials not only affect the assessment of the stream of the gas flow through those materials but they also refer to the loss of pressure energy in that flow. The direct measure of that loss is flow resistances.The results of experimental research upon the assessment of the flow resistances of porous materials with respect to gas flow. The research conducted applied to natural materials with an anisotropic gap-porous structure. The tests were carried out on a gas permeability measuring system, adapted to different shapes of porous material samples. The process issue of the total pressure drop on a porous deposit was considered in the Reynolds number category. The coefficient of flow resistance for anisotropic materials was defined and the value of this coefficient was compared to the gas stream and the total pressure drop on the porous bed was experimentally evaluated.

Simulation programs contain Computational Fluid Dynamics - CFD codes and are a useful tool used for gas flow through porous materials. Conducting numerical simulations allows for detailed analysis of hydrodynamic phenomena. The results of numerical modeling should always be verifiable based on experimental data. Only their compliance with the results of experimental tests is a determinant of the correctness of the applied method. As part of the work, experimental studies of hydrodynamics of gas flow through an isotropic porous material were carried out and numerical simulation for material of the same shape was used. In the CFD modeling Kolmogorov's hypothesis for the transport of kinetic energy of turbulence k and transport of dissipation rate of kinetic energy of turbulence ε was used.

The aim of this work is to evaluate the impact of proposed modifications to conventional capturing and discharging effluents devices in Subsurface Flow Wetland (SSFW) systems. Main modifications proposed consist on extending the influence of the capture and discharge device in such a way that the SSFW width and height are fully covered. Additionally an SSFW prototype was built as pilot including the proposed modifications and the impact of the innovative device is measured based on the efficiency of the Chemical Oxygen Demand (COD) removal as compared to a traditionally built SSFW. The results show that for the innovative device, the COD removal was 10% higher than for the conventional device.

In this paper, we studied and analyzed the behavior of velocity profiles and the distribution of pressures in a pipe connected to an accessory. In this case, it was a 90º elbow fitting in which water flows as an incompressible flow. This study was developed numerically through the fluent program of ANSYS. It is of practical interest in engineering because it is based on results regarding the internal structure of a model of incompressible flows. In the same way, it provides a basis for the proper design and implementation of velocity profiles within an elbow.In the agricultural industry, suitable systems can be implemented for crop irrigation in this way with the results obtained to improve the design in the fluid distribution system.We used a feasible κ-ε turbulence model because this model causes less turbulence than the standard κ-ε model. A mesh independence study was developed, in which it was observed that combining different densities of meshes for the same geometry causes some variables, so a fine meshing was used to carry out the study so that the results are as close to reality as possible.

Expressions for the axial pressure profiles in a cylindrical channel and between parallel plates or a rectangular channel with large aspect ratio, with Maxwell slip gas flow are derived from first principles. The resulting expressions, which only involve the inlet and outlet pressures and the channel dimensions, will be useful in modelling or simulations of channel flows at Knudsen numbers in the range 0.001–0.1, such as in MEMS and NEMS. The expression for a cylindrical channel is validated by deriving from it an expression for the channel mass flow, which is shown to agree with a known expression for the mass flow through cylindrical channels with Maxwell slip flow. The expression for flow between parallel plates is found to agree with the zeroth order relation derived by Arkilic et al. using perturbation analysis. The effect of the accommodation coefficient on the pressure profile in a cylindrical channel is shown.

In ultra-relativistic collisions of heavy ions, the strongly interacting Quark Gluon Plasma (sQGP) is created. The fluid nature of the sQGP was one of the important discoveries of high energy heavy ion physics in the last decades. Henceforth the explosion of this matter may be described by hydrodynamical models. Besides numerical simulations, it is important to study the analytic solutions of the equations of hydrodynamics, as these enable us to understand the connection of the final and initial states better. In this paper we present a perturbative, accelerating solution of relativistic hydrodynamics, on top of a known class of solutions describing Hubble-expansion. We describe the properties of this class of perturbative solutions, and investigate a few selected solutions in detail.

This paper discusses wind tunnel test results aimed at advancing active flow control technology to increase the aerodynamic efficiency of an aircraft during take-off. A model of the outer section of a representative civil airliner wing was equipped with two-stage fluidic actuators between the slat edge and wing tip, where mechanical high-lift devices fail to integrate. The experiments were conducted at a nominal take-off Mach number of M = 0.2. At this incidence velocity, separation on the wing section, accompanied by increased drag, is triggered by the strong slat edge vortex at high angles of attack. On the basis of global force measurements and local static pressure data, the effect of pulsed blowing on the complex flow is evaluated, considering various momentum coefficients and spanwise distributions of the actuation effort. It is shown that through local intensification of forcing, a momentum coefficient of less than c_μ = 0.6% suffices to offset the stall by 2.4°, increase the maximum lift by more than 10%, and reduce the drag by 37% compared to the uncontrolled flow.

Comparisons of power flows and efficiencies in two structurally quite similar cylindrical series-parallel planetary gear systems (PGSs) were performed in three separate parts of this paper. Each of these single degree of freedom (DoF) systems consists of three different 2KH subsystems connected in series-parallel. The main goal was to prove that apart from complex, structurally and dynamically coupled PGSs, there are also complex and only structurally coupled PGSs, in which the phenomenon of power flow circulation inside closed loops does not occur. Such a half-coupled type of PGS is herein called pseudo-coupled. Therefore, Part I discusses in detail the geometry, kinematics and statics of coupled PGSs in order to determine power flow paths. A comparison of the directions of power flow in both types of PGSs can be made in Part II after determining the power flow paths in the second planetary gear in a similar way. The directions of power flow in both types of PGSs were determined in a relatively simple way, thanks to the distinction between active and passive torques and thus active and passive shafts of individual subsystems. The efficiency comparison made in Part III will show whether power circulation has an influence on the efficiency value, at least in these two types of PGSs.

Non-normal mode stability analysis has long been employed to study linear transient characteristics of wall bounded shear flows. However, the application of this approach to investigate shear flows with stratified thermal conditions (STCF) has received limited attention. This paper examines thermal effects on Taylor-Couette flow. The results in this study indicate the possibility of a subcritical bifurcation in stratified Taylor-Couette flow.