Dissociation protocols influence the phenotypes of lymphocyte and myeloid cell populations isolated from the neonatal lymph node.

Frequencies and phenotypes of immune cells differ between neonates and adults in association with age-specific immune responses. Lymph nodes (LN) are critical tissue sites to quantify and define these differences. Advances in flow cytometry have enabled more multifaceted measurements of complex immune responses. Tissue processing can affect the immune cells under investigation that influence key findings. To understand the impact on immune cells in the LN after processing for single-cell suspension, we compared three dissociation protocols: enzymatic digestion, mechanical dissociation with DNase I treatment, and mechanical dissociation with density gradient separation. We analyzed cell yields, viability, phenotypic and maturation markers of immune cells from the lung-draining LN of neonatal and adult mice two days after intranasal respiratory syncytial virus (RSV) infection. While viability was consistent across age groups, the protocols influenced the yield of subsets defined by important phenotypic and activation markers. Moreover, enzymatic digestion did not show higher overall yields of conventional dendritic cells and macrophages from the LN. Together, our findings show that the three dissociation protocols have similar impacts on the number and viability of cells isolated from the neonatal and adult LN. However, enzymatic digestion impacts the mean fluorescence intensity of key lineage and activation markers that may influence experimental findings.

Real-Time Shape Estimation of Hyper-Redundant Flexible Manipulator Using Coiled Fiber Sensors.

The shape of flexible endoscopic surgical robot should be obtained to increase control accuracy and prevent unwanted tissue damage. To estimate the shape of flexible manipulator, space efficiency, cost-effectiveness, system complexity, and ease of calibration should be considered to integrate sensors into the manipulator. In this article, we propose a real-time method to estimate the shape of a hyper-redundant manipulator having embedded coiled fiber sensors. The main advantage of this method is guaranteeing shape recognition even when the manipulator is subjected to an external load. The fiber sensors are highly flexible, compact, and inexpensive, as well as they can functionally measure both compressive and tensile strain of hyper-redundant manipulator. The sensor design was optimized to achieve durability and sensitivity. The numbers of sensor and the placements were determined by the analysis of the kinematics and moment distribution of the manipulator. The accuracy of shape estimation was validated experimentally under both free-loading and loading conditions. The proposed method achieved real-time estimating capability with a mean maximum error of each joint position smaller than 3.54% in free-loading condition and 5.47% in loading condition.

Different outcomes between iso-osmolar and low-osmolar contrast media in acute myocardial infarction with renal impairment.

BACKGROUND: The selection of appropriate contrast media (CM) remains an important issue in terms of renal preservation in patients with acute myocardial infarction (AMI) and renal impairment scheduled for percutaneous coronary intervention (PCI). We compared the clinical outcomes of patients with AMI and renal impairment, depending on the CM type (iso-osmolar CM [IOCM] vs. low-osmolar CM [LOCM]) that was used during PCI. METHODS: From the Convergent Registry of Catholic and Chonnam University for Acute Myocardial Infarction, 3174 post-PCI patients with AMI and renal impairment were subdivided into two groups (IOCM [n = 2101] and LOCM [n = 1073]). RESULTS: Regarding in-hospital clinical outcomes, the IOCM group had a higher peak creatinine (Cr) level and lower "Cr differential" than the LOCM group. A higher proportion of dialysis was noted in the IOCM group. In 30-day clinical outcomes, the IOCM group showed higher incidence of new-onset heart failure (HF) but lower incidence of revascularization than the LOCM group. The differences in in-hospital and 30-day clinical outcomes were attenuated after inverse probability of treatment weighting, except for new-onset HF. All other variables in 30-day clinical outcomes, including all-cause death, non-fatal myocardial infarction, cerebrovascular accidents, stent thrombosis, and any dialysis events, were similar between the two groups. CONCLUSIONS: IOCM use did not prevent future incidence of dialysis compared to LOCM use in AMI patients with renal impairment.

Statistical discrimination using different machine learning models reveals dissimilar key compounds of soybean leaves in targeted polyphenol-metric metabolomics in terms of traits and cultivation.

Soybean (SB) leaves (SLs) contain diverse flavonoids with health-promoting properties. To investigate the chemical constituents of SB and their correlations across phenotypes, growing periods, and environmental factors, a validated separation method for mass detection was used with targeted metabolomics. Thirty-six polyphenols (1 coumestrol, 5 flavones, 18 flavonols, and 12 isoflavones) were identified in SLs, 31 of which were quantified. Machine learning (ML) modelling was used to differentiate between the variety, bean color, growing period, and cultivation area and identify the key compounds responsible for these differences. The isoflavone and flavonol profiles were influenced by the growing period and cultivation area based on bootstrap forest modelling. The neural model showed the best predictive capacity for SL differences among the various ML models. Discriminant polyphenols can differ depending on the ML method applied; therefore, a cautious approach should be ensured when using statistical ML outputs, including orthogonal partial least squares discriminant analysis.

Development of a Digital Healthcare Management System for Lower-Extremity Amputees: A Pilot Study.

The research, which was designed as a "pre- and post-single group" study, included patients with lower-limb amputation and aimed to evaluate the effectiveness of self-directed physical-strength training and cardiovascular exercise using a novel digital healthcare management service three times a week for 12 weeks. Muscle strength, thigh circumference, lipid profile and glycated hemoglobin levels, pulmonary function, quality of life, and physical activity level were evaluated before and after the intervention, while satisfaction was measured after the study. Among the 14 included patients, the proportion of adherence to the physical-strength training and physical-strengthening activity were 85.2% and 75.8%, respectively. The level of satisfaction with the digital healthcare management system was high. Significant changes were observed in the muscle-strength tests (dominant grip power and muscle strength of knee flexion and extension of the intact side), thigh circumference, and glycated hemoglobin levels. Further, the quality-of-life score showed improvement, although without significant differences. Individualized exercise management using the novel digital healthcare management system for lower-limb amputees could induce interest in self-care and promote physical activity and healthy behavior. Through this effect, we can expect a reduction in the incidence of cardiovascular diseases, diabetes mellitus, dyslipidemia, and severe injuries from falling.

Unrestrained Gαi2 Signaling Disrupts Neutrophil Trafficking, Aging, and Clearance.

Neutrophil trafficking, homeostatic and pathogen elicited, depends upon chemoattractant receptors triggering heterotrimeric G-protein Gαißγ signaling, whose magnitude and kinetics are governed by RGS protein/Gαi interactions. RGS proteins typically limit Gαi signaling by reducing the duration that Gαi subunits remain GTP bound and able to activate downstream effectors. Yet how in totality RGS proteins shape neutrophil chemoattractant receptor activated responses remains unclear. Here, we show that C57Bl/6 mouse neutrophils containing a genomic knock-in of a mutation that disables all RGS protein-Gαi2 interactions (G184S) cannot properly balance chemoattractant receptor signaling, nor appropriately respond to inflammatory insults. Mutant neutrophils accumulate in mouse bone marrow, spleen, lung, and liver; despite neutropenia and an intrinsic inability to properly mobilize from the bone marrow. In vitro they rapidly adhere to ICAM-1 coated plates, but in vivo they poorly adhere to blood vessel endothelium. Those few neutrophils that cross blood vessels and enter tissues migrate haphazardly. Following Concanavalin-A administration fragmented G184S neutrophils accumulate in liver sinusoids leading to thrombo-inflammation and perivasculitis. Thus, neutrophil Gαi2/RGS protein interactions both limit and facilitate Gαi2 signaling thereby promoting normal neutrophil trafficking, aging, and clearance.

The oldest Homo erectus buried lithic horizon from the Eastern Saharan Africa. EDAR 7 - an Acheulean assemblage with Kombewa method from the Eastern Desert, Sudan.

Although essential for reconstructing hominin behaviour during the Early Palaeolithic, only a handful of Acheulean sites have been dated in the Eastern Sahara region. This is due to the scarcity of sites for this time period and the lack of datable material. However, recent excavations in the Atbara region (Sudan) have provided unique opportunities to analyse and date Acheulean stone tools. We report here on EDAR 7, part of a cluster of Acheulean and Middle Stone Age (MSA) sites that were recently discovered in the Eastern Desert Atbara River (EDAR) region, located in the Eastern Desert (Sudan) far from the Nile valley. At EDAR 7, a 3.5 metre sedimentary sequence was excavated, allowing an Acheulean assemblage to be investigated using a combination of sedimentology, stone tool studies and optically stimulated luminescence dating (OSL). The site has delivered a complete Acheulean knapping chaine opératoire, providing new information about the Saharan Acheulean. The EDAR 7 site is interpreted as a remnant of a campsite based on the co-occurrence of two reduction modes: one geared towards the production of Large Cutting Tools (LCTs), and the other based on the flaking of small debitage and production of flake tools. Particularly notable in the EDAR 7 assemblage is the abundance of cleavers, most of which display evidence of flake production. Implementation of giant Kombewa flakes was also observed. A geometric morphometric analysis of hand-axes was conducted to verify a possible Late Acheulean assemblage standardisation in the Nubian Sahara. In addition, the analysis of micro-traces and wear on the artefacts has provided information on the use history of the Acheulean stone tools. Sediment analyses and OSL dating show that the EDAR 7 sequence contains the oldest Acheulean encampment remains in the Eastern Sahara, dated to the MIS 11 or earlier. This confirms that Homo erectus occupied the EDAR region during Middle Pleistocene humid periods, and demonstrates that habitable corridors existed between the Ethiopian Highlands, the Nile and the Red Sea coast, allowing population dispersals across the continent and out of it.

Saharan green corridors and Middle Pleistocene hominin dispersals across the Eastern Desert, Sudan.

The Sahara Desert episodically became a space available for hominins in the Pleistocene. Mostly, desert conditions prevailed during the interpluvial periods, which were only periodically interrupted by enhanced precipitation during pluvial or interglacial periods. Responding to Quaternary climatic changes, hominin dispersal was channeled through vegetated corridors. This manuscript introduces a recently discovered group of Acheulean and Middle Stone Age sites far from the Nile Valley in the Eastern Desert (Sudan), referred to as Eastern Desert Atbara River (EDAR). The ∼5 m stratigraphy of the area is divided into three units (Units I-III) bounded by erosion surfaces. Each contains archaeological horizons. The EDAR area has rich surface sites with Acheulean horizons under the surface, singular finds of hand-axes within stratigraphic context in exposures, and large Acheulean sites partly exposed and destroyed by the gold mining activity. Optically stimulated luminescence (OSL) dating of Acheulean and MSA horizons from the EDAR 135 site indicates that the sedimentary deposits with stone artifacts were formed during the Middle Pleistocene between Marine Isotope Stages (MIS) 7 (pluvial) and 6 (interpluvial). Based on the OSL dating from the top of Unit IB, Acheulean artifact-bearing sedimentary deposits from overlying Unit IIA are younger than ca. 231 ka. Unit IA is the oldest Acheulean horizon in the EDAR area, not yet dated but definitively older than ca. 231 ka. An MSA horizon found in fluvial sediment was dated to be between 156 and 181 ka by OSL. The EDAR Pleistocene archaeological sites provide evidence for the presence of additional corridor(s) across Nubia, which connects the early hominin dispersals from the Nile and Atbara River systems to the Red Sea coast.

Real World Comparison of Rivaroxaban and Warfarin in Korean Patients with Atrial Fibrillation: Propensity Matching Cohort Analysis.

Rivaroxaban has emerged as a potential alternative to warfarin for the prevention of thromboembolism in patients with atrial fibrillation (AF). However, there has been concern for the risk of major bleeding, especially in Asian patients. We investigated the efficacy and safety of rivaroxaban compared to warfarin in Korean real world practice. A total of 2,208 consecutive non-valvular AF patients were divided into the Warfarin group (n=990) and the Rivaroxaban group (n=1218). Propensity matched 1-year clinical outcomes were compared (Warfarin, n=804; Rivaroxaban, n=804). The efficacy outcome was defined as stroke/systemic embolism (SE). The safety outcome was major bleeding. The primary net clinical benefit (NCB) was defined as the composite of stroke/SE, major bleeding, and all-cause mortality. Secondary, NCB was defined as the composite of stroke, SE, and major bleeding. Rivaroxaban had the similar efficacy in terms of thromboembolic event prevention [hazard ratio (HR) 0.69, 95% confidence interval (CI) 0.37-1.32, p=0.266] compared to warfarin. Rivaroxaban significantly lowered the risk of major bleeding [HR 0.41, 95% CI 0.22-0.76, p=0.004]. Primary NCB was significantly low in the rivaroxaban group [HR 0.54, 95% CI 0.36-0.81, p=0.003]. Secondary NCB was also low in the rivaroxaban group [HR 0.62, 95% CI 0.40-0.99, p=0.041]. Both rivaroxaban 15 mg and 20 mg groups had similar efficacy and significantly lower risks of major bleeding as well as primary and secondary NCB compared to the warfarin group. In patients with non-valvular AF, rivaroxaban had a similar efficacy to warfarin in Korean real world practice. However, rivaroxaban had better safety and net clinical outcomes compared to warfarin.

Micro Electrode Arrays Fabrication Using Flexible Perfluoroalkoxy Alkane Films.

Perfluoroalkoxy alkane (PFA) film-based microelectrode arrays were fabricated in monolithic structure with pretreatment of plasma on the surface followed by patterning gold electrode array without additional adhesion metal layer. The encapsulation of patterned fluoropolymer layer was thermally laminated at the temperature of under its melting point, interleaved between Polytetrafluoroethylene (PTFE) sheets which indicate formation of micro-corrugated pattern. The fabricated device was optically opaque, however, transparent in solution due to the diffraction of light. Packaged substrate was easily processable with focused ion beam laser machine in high resolution. According to excellent reliability, PFA based micro gold electrode can be applied in a variety of biomedical application such as artificial retina, neural sensors with chronic implantation.

Characterization of morphological changes of B16 melanoma cells under natural killer cell attack.

Natural killer (NK) cell killing of melanoma cells involves perforin-mediated delivery of granzymes from NK cells to cancer cells; however, how melanoma cells die remains poorly characterized. Here, we examined the dying process of melanoma cells by using time-lapse imaging. Upon contact with NK cells, B16-F10 cells rounded and most of them showed membrane rupture (98 min); however, B16 parent cells showed writhing and delayed membrane rupture (235 min). This morphological difference depended on the expression levels of myosin regulatory light chain 9 (MYL9) but not activating ligands (CD112, CD155, Rae-1, and MULT-1), SPI, FasL, or PD-L1. Taken together, our data show that melanoma cells show two distinct types of morphological changes upon contact with NK cells and suggest that a strategy to decrease MYL9 expression by melanoma cells may improve the efficacy of NK cell-based immunotherapy.

CXCR3-deficient natural killer cells fail to migrate to B16F10 melanoma cells.

Natural killer (NK) cells eliminate cancer cells in a contact-dependent manner. However, how NK cells find cancer cells remain unclear. Here, using time-lapse imaging, we investigated how individual NK cells migrate toward cancer cells. Although naïve B16F10 cancer cells produce low levels of chemokines, IFN-γ-treated B16F10 cells secreted high levels of CXCL10, low levels of CCL5, but did not secrete CCL2, CCL7, or CXCL12. Wild-type NK cells migrated well toward cancer cells and killed them, whereas NK cells deficient in CXCR3 did not. CXCR3-deficient NK cells also showed slower migration speed than did wild-type NK cells. Taken together, our data show that NK cells find cancer cells, at least in part, by sensing CXCL10 produced by cancer cells and suggest that a strategy to increase CXCL10 secretion by cancer cells may improve the efficacy of NK cell-based immunotherapy.

Effect of a Combination of Prednisone or Mycophenolate Mofetil and Mesenchymal Stem Cells on Lupus Symptoms in MRL.Faslpr Mice.

The combination of immunosuppressants and mesenchymal stem cells (MSCs) is a promising therapeutic strategy for systemic lupus erythematosus, since this approach reduces doses of immunosuppressants while maintaining the same therapeutic outcome. However, it is unavoidable for MSCs to be exposed to immunosuppressants. Here, we examined the combination effect of prednisone (PD) or mycophenolate mofetil (MMF) and MSCs. We showed that PD or MMF in combination with MSCs showed better therapeutic effect than single therapy in lupus-prone MRL.Faslpr mice, as assessed by using the following readouts: prolongation of survival, decrease in anti-dsDNA and total IgG levels in serum, decrease in cytokine gene expression in spleen cells, and decrease in inflammatory cell infiltration into the kidney. In vitro, immunosuppressants and MSCs inhibited T cell proliferation in a synergistic manner. However, immunosuppressants did not affect MSC viability and functions such as TGF-ß1 and PGE2 production, migration, and immunosuppressive capacity. In summary, our study demonstrates that a combination of immunosuppressants and MSCs is a good strategy to reduce the side effects of PD and MMF without the loss of therapeutic outcome.

RIDDLE: Race and ethnicity Imputation from Disease history with Deep LEarning.

Anonymized electronic medical records are an increasingly popular source of research data. However, these datasets often lack race and ethnicity information. This creates problems for researchers modeling human disease, as race and ethnicity are powerful confounders for many health exposures and treatment outcomes; race and ethnicity are closely linked to population-specific genetic variation. We showed that deep neural networks generate more accurate estimates for missing racial and ethnic information than competing methods (e.g., logistic regression, random forest, support vector machines, and gradient-boosted decision trees). RIDDLE yielded significantly better classification performance across all metrics that were considered: accuracy, cross-entropy loss (error), precision, recall, and area under the curve for receiver operating characteristic plots (all p < 10-9). We made specific efforts to interpret the trained neural network models to identify, quantify, and visualize medical features which are predictive of race and ethnicity. We used these characterizations of informative features to perform a systematic comparison of differential disease patterns by race and ethnicity. The fact that clinical histories are informative for imputing race and ethnicity could reflect (1) a skewed distribution of blue- and white-collar professions across racial and ethnic groups, (2) uneven accessibility and subjective importance of prophylactic health, (3) possible variation in lifestyle, such as dietary habits, and (4) differences in background genetic variation which predispose to diseases.

Cd226-/- natural killer cells fail to establish stable contacts with cancer cells and show impaired control of tumor metastasis in vivo.

CD226 is an activating receptor expressed on natural killer (NK) cells, CD8+ T cells, and other immune cells. Upon binding to its ligands expressed on target cells, CD226 activates intracellular signaling that triggers cytokine production and degranulation in NK cells. However, the role of CD226 in contact dynamics between NK and cancer cells has remained unclear. Our time-lapse images showed that individual wild-type CD226+ NK cells contacted B16F10 melanoma cells for 23.7 min, but Cd226-/- NK cells only for 12.8 min, although both NK cell subsets showed equal contact frequency over 4 h. On the surface of B16F10 cells, CD226+ cells stayed at the same site with oscillating movement (named stable contact), while Cd226-/- NK cells moved around at a velocity of 4 µm/min (named unstable contact). Consequently, Cd226-/- NK cells did not kill B16F10 cells in vitro and did not inhibit their metastasis into the lung in vivo. Taken together, our data demonstrate that CD226 enables prolonged stable interaction between NK and cancer cells, which is needed for efficient killing of cancer cells.

CCL2 deficient mesenchymal stem cells fail to establish long-lasting contact with T cells and no longer ameliorate lupus symptoms.

Systemic lupus erythematosus (SLE) is a multi-organ autoimmune disease characterized by autoantibody production. Mesenchymal stem cells (MSCs) ameliorate SLE symptoms by targeting T cells, whereas the mechanisms of their efficacy remain incompletely understood. In this study, we show that transfer of human MSCs increased MRL.Faslpr mouse survival, decreased T cell infiltration in the kidneys, and reduced T cell cytokine expression. In vitro, allogeneic mouse MSCs inhibited MRL.Faslpr T cell proliferation and cytokine production. Time-lapse imaging revealed that MSCs recruited MRL.Faslpr T cells establishing long-lasting cellular contacts by enhancing T cell VCAM-1 expression in a CCL2-dependent manner. In contrast, CCL2 deficient MSCs did not induce T cell migration and VCAM-1 expression, resulting in insufficient cell-cell contact. Consequently, CCL2 deficient MSCs did not inhibit IFN-γ production by T cells and upon transfer no longer prolonged survival of MRL.Faslpr mice. Taken together, our imaging study demonstrates that CCL2 enables the prolonged MSC-T cell interactions needed for sufficient suppression of autoreactive T cells and helps to understand how MSCs ameliorate symptoms in lupus-prone MRL.Faslpr mice.

Fabrication of Localized Surface Plasmon Resonance Sensor Based on Optical Fiber and Micro Fluidic Channel.

This paper proposes Fiber-Optic Localized Surface Plasmon Resonance (FO LSPR) sensor combined with a micro fluidic channel, which enables continuous supply of fluid for bio-reaction. The proposed method prevents degradation of the sensing performance due to changes in measurement conditions. The feasibility of the FO LSPR sensor with a micro fluidic channel was demonstrated by computational fluid dynamics (CFD) simulation. Also, the proposed method was assessed by measuring the output intensity of the FO LSPR sensor at various refractive index solutions. Finally, a prostate-specific antigen (PSA) immunoassay was performed to evaluate the possibility of the fabricated sensor system as a biosensor.

Cytokine-induced killer cells hunt individual cancer cells in droves in a mouse model.

Cytotoxicity of cytokine-induced killer (CIK) cells depends mainly on their encounters with target cells, but how many CIK cells are required to kill an individual cancer cell is unknown. Here we used time-lapse imaging to quantify the critical effector cell number required to kill an individual target cell. CIK cells killed MHC-I-negative and MHC-I-positive cancer cells, but natural killer (NK) cells destroyed MHC-I-negative cells only. The average threshold number of CIK cells required to kill an individual cancer cell was 6.7 for MHC-I-negative cells and 6.9 for MHC-I-positive cells. That of NK cells was 2.4 for MHC-I-negative cells. Likely due to the higher threshold numbers, killing by CIK cells was delayed in comparison with NK cells: 40% of MHC-negative target cells were killed after 5 h when co-cultured with CIK cells and after 2 h with NK cells. Our data have implications for the rational design of CIK cell-based immunotherapy of cancer patients.

A Korean Multi-Center Survey about Warfarin Management before Gastroenterological Endoscopy in Patients with a History of Mechanical Valve Replacement Surgery.

BACKGROUND: Guidelines for esophagogastroduodenoscopy (EGD) in the West allow the continued use of warfarin under therapeutic international normalized ratio (INR) level. In Korea, no guidelines have been issued regarding warfarin treatment before EGD. The authors surveyed Korean cardiac surgeons about how Korean cardiac surgeons handle warfarin therapy before EGD using a questionnaire. Participants were requested to make decisions regarding the continuation of warfarin therapy in two hypothetical cases. METHODS: The questionnaire was administered to cardiac surgeons and consisted of eight questions, including two case scenarios. RESULTS: Thirty-six cardiac surgeons at 28 hospitals participated in the survey, and 52.7% of the participants chose to stop warfarin before EGD in aortic valve replacement patients without risk factors for thromboembolism. When the patient's INR level was 2, 31% of the participants indicated that they would choose to continue warfarin therapy. For EGD with biopsy, 72.2% of the participants chose warfarin withdrawal, and 25% of the participants chose heparin replacement. In mitral valve replacement patients, 47.2% of the participants chose to discontinue warfarin, and 22.2% of the participants chose heparin replacement. For EGD with biopsy in patients with a mitral valve replacement, 58.3% of the participants chose to stop warfarin, and 41.7% of the participants chose heparin replacement. CONCLUSION: This study demonstrated that attitudes regarding warfarin treatment for EGD are very different among Korean surgeons. Guidelines specific to the Korean population are required.

Curdlan activates dendritic cells through dectin-1 and toll-like receptor 4 signaling.

Curdlan, a ß-1,3-glucan isolated from Alcaligenes faecalis, is an agonist of dectin-1 in various immune cells, including dendritic cells (DCs). However, whether curdlan also activates DCs through other receptors remains unknown. In this study, we found that curdlan activates DCs through dectin-1 and toll-like receptor 4 (TLR4). Curdlan increased the expression levels of surface molecules (CD40, CD80, CD86, and MHC-I/II), the production of cytokines (IL-12, IL-1ß, TNF-α, and IFN-ß), migration toward MIP-3ß, and allogeneic T cell stimulation activity of DCs. Curdlan increased the phosphorylation of Syk, Raf-1, Akt, MAPKs, IKK, and NF-κB p65 in DCs. However, curdlan only slightly activated DCs transfected with small interfering RNAs against dectin-1 or TLR4 and C3H/HeJ DCs, which have non-functional TLR4, in comparison with control DCs. Curdlan increased antitumor activity of DCs in a syngeneic tumor model. In summary, our data show that curdlan activates DCs through dectin-1 and TLR4 signaling and the combination of curdlan and DCs efficiently inhibit tumor growth in mice.