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Background Single-cell (sc) sequencing performs unbiased profiling of individual cells and enables evaluation of less prevalent cellular populations, often missed using bulk sequencing. However, the scale and the complexity of the sc datasets poses a great challenge in its utility and this problem is further exacerbated when working with larger datasets typically generated by consortium efforts. As the scale of single cell datasets continues to increase exponentially, there is an unmet technological need to develop database platforms that can evaluate key biological hypotheses by querying extensive single-cell datasets. Large single-cell datasets like Human Cell Atlas and COVID-19 cell atlas (collection of annotated sc datasets from various human organs) are excellent resources for profiling target genes involved in human diseases and disorders ranging from oncology, auto-immunity, as well as infectious diseases like COVID-19 caused by SARS-CoV-2 virus. SARS-CoV-2 infections have led to a worldwide pandemic with massive loss of lives, infections exceeding 7 million cases. The virus uses ACE2 and TMPRSS2 as key viral entry associated proteins expressed in human cells for infections. Evaluating the expression profile of key genes in large single-cell datasets can facilitate testing for diagnostics, therapeutics, and vaccine targets, as the world struggles to cope with the on-going spread of COVID-19 infections. Main body In this manuscript we describe REVEAL: SingleCell, which enables storage, retrieval, and rapid query of single-cell datasets inclusive of millions of cells. The array native database described here enables selecting and analyzing cells across multiple studies. Cells can be selected using individual metadata tags, more complex hierarchical ontology filtering, and gene expression threshold ranges, including co-expression of multiple genes. The tags on selected cells can be further evaluated for testing biological hypotheses. One such example includes identifying the most prevalent cell type annotation tag on returned cells. We used REVEAL: SingleCell to evaluate the expression of key SARS-CoV-2 entry associated genes, and queried the current database (2.2 Million cells, 32 projects) to obtain the results in Conclusion In this paper, we introduce the REVEAL: SingleCell database that addresses immediate needs for SARS-CoV-2 research and has the potential to be used more broadly for many precision medicine applications. We used the REVEAL: SingleCell database as a reference to ask questions relevant to drug development and precision medicine regarding cell type and co-expression for genes that encode proteins necessary for SARS-CoV-2 to enter and reproduce in cells.

Neural stem cells (NSCs) are multipotent and are considered ideal source for regenerating damaged neural cells for neurological disorders. During culture of NSCs, both the measurement and the evaluation of their differentiation potential are important to maintain stable quality-assured NSCs for regenerative treatments since the rate of differentiation into certain lineages from NSCs is still not fully controllable. However, conventional cell evaluation techniques using biological molecular are still invasive, costly, and time-consuming. Therefore, a non-invasive, low-cost, and rapid cell evaluation method is required to expand the possibilities of regenerative therapy, especially in the facilities that produce cells for therapy. To address these such technological limitations in non-invasive cell evaluation, we propose the efficacy of computer-aided morphology-based prediction of potentials of stem cells by using multiple and time-course morphological parameters from phase-contrast microscopic images combined with experimentally determined differentiation potentials. In this work, we quantified the morphological parameters of NSCs during three types of differentiation culture and investigated two applications with NSCs: (i) evaluation of their differentiation type and (ii) early prediction of neural differentiation rate. Our data demonstrate that it is possible to non-invasively evaluate neural differentiation types and quantitatively predict future differentiation rates by using morphological information from the first 4 days. Our findings indicate the potential application of morphology-based non-invasive evaluation for optimizing effective differentiation protocols, screening of compounds to mediate NSC differentiation, and quality maintenance of regenerative medicine products.

In this research, thermosensitive liposomes (t-L) containing juglone were prepared with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and cholesterol by aether injection method. Morphological characteristics, encapsulation efficiency (EE), particle size, zeta potential, in vitro release, and cell viability of t-L were investigated and compared with those of conventional liposomes (c-L). Results indicated that t-L are multilamellar vesicles with lower negative charge, larger particle diameter, and higher EE than c-L. Moreover, in vitro juglone release from t-L was temperature dependent. Up to 60 % of the loaded juglone was released from t-L in 20 min when environmental temperature was increased from 37 °C to 42 °C; by contrast, >60 % of the drug remained inside for 24 h at 37 °C. Furthermore, MTT assay results revealed that t-L significantly increased the inhibitory effect on HepG2 cell growth and proliferation when these cells were exposed to hyperthermia; therefore, t-L could be applied for targeted therapy.

As the number of available cell types grows, it becomes necessary to develop more effective ways to optimize the cell-culture medium for each cell line and culture condition. However, because of the vast number of parameters that must be decided, such as the combination of components, optimization is both laborious and costly. Microdevices are a cost-effective way to perform such evaluations because they use only a small volume of media and enable high-throughput analyses. However, assays performed in microdevices are themselves minimized, and each assay unit (well/chamber) commonly contains an insufficient number of cells for comprehensive evaluations such as gene-expression or flow-cytometry analyses. To address this issue, we introduced image-based analysis in conjunction with microdevice assays; this approach allows quantification of every cell in each assay unit. To quantitatively profile differences in cellular behaviors in a microdevice under different culture media conditions, we developed a non-staining image-based analysis method that utilizes cellular morphology. Our approach combines the structural advantages of microdevices, which can increase the stability of images, and the quantitative advantages of an image-based cell evaluation technique that utilizes time-course population change in several morphological features. Our results demonstrate that cellular changes due to small alterations in the concentration of serum in medium or differences in the basal medium can be profiled using only microscopic images.

Purpose To optimize methods of preparing donor cornea tissue for Descemet's stripping automated endothelial keratoplasty (DSAEK), we compared five experimental conditions with different internal pressures and cutting systems. Methods The artificial anterior chamber internal pressure (IP) was set at 100 or 200 mm Hg. The microkeratome cut was performed with or without an artificial chamber pressurizer (ACP), using a CBm turbine (CBm) or one use-plus automated (OUP-A). Thirty human research corneas were divided into five groups, and compared after the cut with donor tissue quality parameters, including cutting depth, graft uniformity, cell evaluation, and smoothness of the stromal surface. Results The smallest variation in mean cut depth was observed in the condition, which had IP of 200 mm Hg used ACP and OUP-A. In experimental groups cut using CBm, significantly more consistent thicknesses were made at an IP of 200 than 100 mm Hg. There were no statistically significant differences among the groups in either endothelial cell density or cell viable assay results after cuts. Using an IP of 200 mm Hg with ACP and CBm produced the roughest stromal surface, and the roughness grading scores showed a positive correlation with the percentage of cut depth. Conclusions An IP of 200 mm Hg was the best setting for DSAEK grafts with high predictability of cut depth and uniformity of graft thickness without endothelial cell damage. Translational relevance For successful DSAEK, it is recommended that a set internal pressure of 200 mm Hg be used during microkeratome cutting for donor tissue preparation.

BACKGROUND: Altered microglial markers and morphology have been demonstrated in patients with schizophrenia in post-mortem and in vivo studies. However, it is unclear if changes are due to antipsychotic treatment. AIMS: Here we aimed to determine whether antipsychotic medication affects microglia in vivo. METHODS: To investigate this we administered two clinically relevant doses (0.05 mg n=12 and 2.5 mg n=7 slow-release pellets, placebo n=20) of haloperidol, over 2 weeks, to male Sprague Dawley rats to determine the effect on microglial cell density and morphology (area occupied by processes and microglial cell area). We developed an analysis pipeline for the automated assessment of microglial cells and used lipopolysaccharide (LPS) treatment ( n=13) as a positive control for analysis. We also investigated the effects of haloperidol ( n=9) or placebo ( n=10) on the expression of the translocator protein 18 kDa (TSPO) using autoradiography with [3H]PBR28, a TSPO ligand used in human positron emission tomography (PET) studies. RESULTS: Here we demonstrated that haloperidol at either dose does not alter microglial measures compared with placebo control animals ( p > 0.05). Similarly there was no difference in [3H]PBR28 binding between placebo and haloperidol tissue ( p > 0.05). In contrast, LPS was associated with greater cell density ( p = 0.04) and larger cell size ( p = 0.01). CONCLUSION: These findings suggest that haloperidol does not affect microglial cell density, morphology or TSPO expression, indicating that clinical study alterations are likely not the consequence of antipsychotic treatment. The automated cell evaluation pipeline was able to detect changes in microglial morphology induced by LPS and is made freely available for future use.

Coulombic efficiency and capacity retention are some of the main issues with alloy anodes in lithium-ion batteries, and the use of electrolyte additives is a viable approach to overcome these problems. Accordingly, the effects of three electrolyte additives, viz., vinylene carbonate (VC), fluoroethylene carbonate (FEC), and vinylethylene carbonate (VEC), on the performance of a promising alloy anode material Cu 2 Sb–Al 2 O 3 –C have been investigated. With an aim to rapidly assess the effects of the electrolyte additives, symmetric cells fabricated with Cu 2 Sb–Al 2 O 3 –C were used. Symmetric cell testing showed results similar to those of conventional half-cell testing in as little as one-fifth of the time, illustrating that symmetric cell testing is a valuable tool for optimization the performance of battery materials. Among the various additives and concentrations investigated, 2% VEC was found to offer the best cell performance with the Cu 2 Sb–Al 2 O 3 –C nanocomposite anode.

In this paper, we discuss a development of control and sensing platform for an on-chip single cell evaluation by magnetically driven microtool (MMT). The design and fabrication of the MMT is shown. Through the basic experiments, the advantage of the proposed platform, the performance of the position control (positioning accuracy: 30 µm), and the force sensing (sensing accuracy: 100 µN) of the MMT are confirmed.

An integrated flow determination algorithm is proposed to calculate the spatial distribution of the topographic index to the channel network. The advantages of a single flow direction algorithm and other multiple flow direction schemes are selectively considered in order to address the drawbacks of existing algorithms. A spatially varying flow apportioning factor is introduced to distribute the contributing area from upslope cells to downslope cells. The channel initiation threshold concept is expanded and integrated into a spatially distributed flow apportioning algorithm to delineate a realistic channel network. The functional relationships between the flow apportioning factors and the expanded channel initiation threshold (ECIT) are developed to address the spatially varied flow distribution patterns considering the permanent channel locations. A genetic algorithm (GA) is integrated into the spatially distributed flow apportioning algorithm (SDFAA) with the objective function of river cell evaluation. An application of a field example suggests that the spatially distributed flow apportioning scheme provides several advantages over the existing approaches; the advantages include the relaxation of overdissipation problems near channel cells, the connectivity feature of river cells and the robustness of the parameter determination procedure over existing algorithms. Copyright © 2004 John Wiley & Sons, Ltd.

An objective evaluation of semen is warranted to assess the canine male fertility and to select appropriate techniques and extenders for semen preservation. With conventional microscopic evaluation, the subjectivity of the analysis makes any comparison of results difficult. In the present study, we validated the Hamilton Thorn computer-aided semen analyzer (HTR-IVOS10 analyzer) for objective assessment of canine semen. A description of fertile canine motility parameters using this analyzer is reported. Semen analysis at 38 degreesC is found to be more optimal and accurate than 30 degreesC. The Makler chamber was preferred to the Cell-vu, which induced a decrease of all semen motility parameters. The repeatability of the measures was good with intra-and inter-assay coefficients of variation below 10% and 20%, respectively, for most of measured parameters. An overestimation of semen concentration, increasing with dilution of semen, was observed when HTR-IVOS10 results were compared with the classical manual Makler cell evaluation. A significant decrease in semen motility parameters was recorded when high semen dilutions were used. Generated from the analysis of 42 mature fertile male beagle dogs, a description of semen motility parameters using the CASA system is presented to serve as reference when comparing Beagle ejaculates both in clinical and experimental studies.

Cell formation (CF) consists of identifying machine groups (MGs) and part families (PFs). Many CF procedures use a part machine matrix as an input and attempt to obtain a block diagonal form. A perfect diagonalization of the part machine matrix to form exclusive PFs and MGs is not possible in many instances. Considering alternate routeings (i.e. alternate plans for the parts and additional copies of machines) improves this diagnonalization. This aspect has not been adequately dealt with in literature. Moreover, existing CF procedures consider indirect measures such as similarity indices, rank order, bond energy etc., that may not obtain a good block diagonalization of the part machine matrix. Also these procedures decouple cell formation and cell evaluation procedures. In this paper a non-linear integer programming model is developed for CF to identify PFs and MGs simultaneously considering alternative routeings. The model combines the evaluation procedure by considering the minimization of a weighted sum...

BACKGROUND AND OBJECTIVES The aim of this study was first, to analyze the post-thaw progenitor assays usually performed on peripheral blood stem cell autografts and second, to achieve standardization with improved flow cytometric and CFU-GM assays. MATERIALS AND METHODS In the first part of the study (n=79), recovery and Intraclass Correlation Coefficient (ICC) of total nucleated cells, CD34 and CFU-GM were analyzed before and after cryopreservation. In the second part (n=20), evaluation methods were modified : the washing step was suppressed in the flow cytometric method and 500 CD34 were plated compared to 4×10(4) total nucleated cells in the CFU-GM assay. The recovery rates were analyzed and the CFU-GM results were regarded as reliable when 30-100 colonies were observed, according to the manufacturer recommendation. RESULTS The analysis of the first part showed an ICC that was perfect for total nucleated cells (0.93), substantial for CD34 (0.67) and fair for CFU-GM (0.25). Median CD34 recovery was 112.6% (29.9-222%). The CFU-GM median recovery was 31.7% (0.19-142%) leading to reliable results for 27 grafts. In the second part, the median CD34 recovery was 85.75% (54-99%). No recovery over 100% was observed. The CFU-GM assay led to 18 out of 20 evaluable autografts when 500 CD34 were seeded, compared to 10 out of 20 when total nucleated cell were seeded. CONCLUSION Avoiding cell washing in the flow cytometric method limited the overestimate of the CD34 percentage. Plating 500 thawed CD34 improved reliability of the results and allowed a better standardization of the assay.

To study the sensitivity of laser flare photometry (LFP) in monitoring anterior chamber inflammation by correlating LFP measurements with slit-lamp evaluation of aqueous cells in HLA-B27-related uveitis in a prospective trial. Slit-lamp cell evaluation was correlated with LFP-measured flare in a masked fashion in HLA-B27-related uveitis patients receiving standard topical therapy. At the time of 50 and 90% LFP flare reduction, the corresponding reduction of cells was recorded and statistically compared using the sign test. Forty-three episodes (in 43 patients) of acute anterior HLA-B27-related uveitis were included. LFP flare reduction and slit-lamp cell reduction were strongly correlated. LFP was significantly more sensitive for both 50% (P = 0.001) and 90% (P = 0.02) LFP flare reduction in assessing the decrease of anterior chamber inflammation. LFP was superior to slit-lamp cell evaluation in monitoring anterior chamber inflammation in uveitis. Flare, becoming a quantitative parameter when measured by LFP, rather than cells, should be considered the gold standard to measure anterior chamber inflammation in uveitis.

An overview of both experimental and theoretical studies of cell electrophoresis mobility (EPM) over the past fifty years and the relevance of cell EPM measurement are presented and discussed from the viewpoint of exploring the potential use of cell EPM as an index of the biological condition of cells. Physical measurements of the optical and/or electrical properties of cells have been attracting considerable attention as noninvasive cell-evaluation methods that are essential for the future of cell-based application technologies such as cell-based drug screening and cell therapy. Cell EPM, which can be measured in a noninvasive manner by cell electrophoresis, reflects the electrical and mechanical properties of the cell surface. Although the importance of cell EPM has been underestimated for a long time, mostly owing to the technical difficulties associated with its measurement, recent improvements in measurement technology using microcapillary chips have been changing the situation: cell EPM measurement has become more reliable and faster. Recent studies using the automated microcapillary cell electrophoresis system have revealed the close correlation between cell EPM and important biological phenomena including cell cycle, apoptosis, enzymatic treatment, and immune reaction. In particular, the converged EPM distribution observed for synchronized cells has altered the conventional belief that cell EPMs vary considerably. Finding a new significance of cell EPM is likely to lead to noninvasive cell evaluation methods essential for the next-generation of cell engineering.

Animal derived protein hydrolysates are commonly used as a complex supplement for cell culture media because of their well known growth promoting properties. In an approach to find components being responsible for this effect we fractionated a protein hydrolysate by solid phase extraction (SPE) on a C18-matrix. After lyophilization and further preparation the fractions were tested for their proliferation enhancing ability with a hybridoma cell line in a serum-free DMEM/F12 medium supplemented with human serum albumin and transferrin. We could demonstrate a growth promoting effect caused by a rather hydrophilic fraction. In further investigations preparative High Performance Liquid Chromatography (HPLC) was used to produce growth promoting fractions of less complexity for cell evaluation and analytical HPLC.

BACKGROUND: The accuracy of synovial fluid (SF) white cell count (WCC) and polymorphonuclear (PMN) cell evaluation for predicting prosthetic joint infection (PJI) at the total hip arthroplasty (THA) or total knee arthroplasty (TKA) site is unknown. Therefore, we performed a meta-analysis to summarize the diagnostic validity of SF-WCC and SF-PMN for diagnosing PJI. METHODS: The MEDLINE, EMBASE, and OVID databases were searched for studies that had evaluated the diagnostic validity of SF-WCC and SF-PMN between January 1990 and May 2013. Meta-analysis methods were used to pool sensitivity, specificity, diagnostic odd ratios (DORs), the area under the receiver-operating characteristic curve (AUC), positive likelihood ratios (PLR), negative likelihood ratios (NLR), and post-test probability. We also conducted heterogeneity, publication bias, subgroup, and meta-regression analyses. RESULTS: Fifteen articles (15 SF-WCC and 14 SF-PMN) that included a total of 2787 patients fulfilled the inclusion criteria and were considered for analysis. The pooled sensitivity and specificity for PJI detection was 0.88 (95% confidence intervals [CI], 0.81-0.93) and 0.93 (95% CI, 0.88-0.96) for SF-WCC and 0.90 (95% CI, 0.84-0.93) and 0.88 (95% CI, 0.83-0.92) for SF-PMN, respectively. The AUC was 0.96 for SF-WCC and 0.95 for SF-PMN. PLR and NLR were 13.3 and 0.13 for SF-WCC, and 7.6 and 0.12 for SF-PMN, respectively. There was no evidence of publication bias. Low-clinical-scenario (pre-test probability, 20%) post-test probabilities were 3% for both negative SF-WCC and SF-PMN results. The subgroup analyses indicated that the sensitivity/specificity of THA were 0.73/0.96 for SF-WCC and 0.85/0.83 for SF-PMN, whereas those of TKA were 0.90/0.91 for SF-WCC and 0.90/0.88 for SF-PMN. We also found that collection of SF-WCC preoperatively had a higher sensitivity than that obtained intraoperatively (0.91 vs. 0.77). CONCLUSIONS: SF-WCC and SF-PMN have an adequate and clinically acceptable diagnostic value for detecting PJI, particularly after TKA.

In this work we report the development of the tri-functionalized mesoporous silica nanoparticles (MSNs) for use as theranostic compounds that orchestrate the trio of imaging, target and therapy in a single particle. The MSNs are functionalized in sequence with (1) contrast agents that enable traceable imaging of particle targeting, (2) drug payloads for therapeutic intervention and, (3) biomolecular ligands for highly-targeted particle delivery. Traceable imaging of nanoparticles was accomplished by directly incorporating a near-infrared (NIR) fluorescent contrast agent, ATTO647N, into the silica framework of MSNs, to exploit the relative transparency of most tissues at NIR wavelengths and maximize MSN surface area available for the subsequent conjugating drugs and targeting ligands. An oxygen-sensing, palladium-porphyrin based photosensitizer (Pd-porphyrin; PdTPP) was incorporated into the MSN's nanochannels, to enable photodynamic therapy (PDT). cRGDyK peptides, tiling the outermost surfaces of MSNs, were used for targeting the overexpressed αvβ3 integrins of cancer cells, and to ensure the internalization of the photosensitizer PdTPP. In vitro cell evaluation of the theranostic platform demonstrated not only excellent targeting specificity and minimal collateral damage, but highly potent therapeutic effect as well.

The beam performance of the electron optics proof of concept (POC) multicolumn-cell (MCC) system has been evaluated. The two beams at the near center of a 4×4 layout with 25 mm pitch show good uniformity and low interference. The MCC system is imaged with a variable-shaped beam and character projection electron optical system having a vector scan beam deflection and a “write-on-the-fly” stage motion control. Cost performance analysis gives the most effective number of column cells in the MCC system.

Initial cell evaluation on alginate/hydroxyapatite block was investigated. Sodium alginate with 1, 3 and 5% concentration was obtained via neutral extraction of locally obtained brown seaweed, Sargassumpolycystum. Commercially available hydroxyapatite (HAp) powder was pressed uniaxially at 3 MPa to obtain the HAp block. The HAp block was then sintered at 900C. The sintered HAp block was then immersed in the sodium alginate solution at different concentration for 24 hours under vacuum condition. Morphological observations show that normal cell growth was observed on alginate/HAp blockafter post treatment for day 1 and 2. However, the cell starts to show some distinct morphological changes when compared to the control cells for day 5 and 7. Cell viability assay results shows that a consistent cell growth was obtained with HAp block incorporated with 3 and 5% sodium alginate. While HAp block without the incorporation of sodium alginate and HAp block incorporated with 1% sodium alginate concentration shows inconsistent cell growth. Initial cell evaluation results suggest that alginate/HAp block shows no toxicity on cell attachment and proliferation.

Operational characteristics of the LANL electrorefining cell, a modified LANL electrorefining cell, and an advanced electrorefining cell (known as the CRAC cell) were determined. Average process yields achieved were: 75% for the LANL cell, 82% for the modified LANL cell, and 86% for the CRAC cell. All product metal from the LANL and modified LANL cells was within foundry specifications. Metal from one run in the CRAC cell exceeded foundry specifications for tantalum. The LANL and modified LANL cells were simple in design and operation, but product separation was more labor intensive than with the CRAC cell. The CRAC cell was more complicated in design but remained relatively simple in operation. A decision analysis concluded that the modified LANL cell was the preferred cell. It was recommended that the modified LANL cell be implemented by the Plutonium Recovery Project at Rocky Flats and that development of the CRAC cell continue. 8 refs., 22 figs., 12 tabs.

The credit of introducing specular microscopy for the study of the corneal endothelium goes to D. Maurice. However, it was Vogt, who showed in 1931 that the corneal endothelium could well be visualized with the conventional slit-lamp. Laing, Sandstrom and Leibowitz on one side, Bourne, Mc Carey and Kaufman on the other side modified the original system for the human in vivo cell evaluation. Several studies have since been published by different groups on the problem of endothelial cell loss after routine cataract operation and after implantation of intraocular artifical lenses. L.W. Hirst, R.C. Snip, W.I. Starck and A.E. Maumenee recently wrote a paper on the same subject and came to the final conclusion, that all the figures published so far should be viewed in the light of the problems of the photography used. The analysis of the data obtained is certainly open to many questions.

This paper discusses the test results of an automatic, real-time Engine Performance Monitoring System (EPMS) which was flight tested on-board a commercial airline B-707-321B aircraft. EPMS employed the steady-state technique used in test cells to determine the “health” of the operating jet engine. Additional techniques and applications being developed by Emerson Electric Co., such as the portable ground based jet engine analyzer and the NAVY Jet Engine Transient Health Analyses Study, are also discussed.Copyright © 1971 by ASME