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Indonesia is the center of the diversity of bananas that the wild relatives co-exist with the edible bananas or cultivars. The ploidy information of cultivars and wild relatives is imperative for germplasm management for collection and cultivation as well as breeding programs. However, only a few numbers of Indonesian bananas have been empirically confirmed using cytogenetic approaches such as either chromosome counting or flow cytometry. Hence, such approaches are needed to disclose the genetic properties of banana that grow in Indonesia precisely. Flow cytometry is a method that offers a high throughput and is efficient in determining the ploidy level of a plant. In this study, we determined the ploidy level of banana cultivars and their wild relatives that we collected during period 2009 to 2019. In total, we analyzed 257 accession samples. From this analysis, we observed 69 samples were diploid, 168 accessions were triploid, and 21 accessions were tetraploid. Among wild bananas, 14 wild Musa acuminata accessions and four M. balbisiana accessions were diploid. Interestingly, 'Siem', 'Barangan' and 'Kepok'cultivars in general, described as triploid bananas, were determined as tetraploid. The result indicates that the ploidy evaluation of Indonesian bananas needs to be continued to disclose germplasm potential for bananas breeding programs in the future. The advent of Indonesian bananas' ploidy level information not only to understand the genetics of bananas but also will help breeders to select parents for breeding diploid, triploid or tetraploid bananas. [ABSTRACT FROM AUTHOR]

The objective of this study was to establish in vitro propagation system of Merkeron napier grass (Pennisetum purpureum) via multiple-shoot clumps formation. Shoot apical meristem as explants were isolated from shoot-tillers and cultured in vitro on a solid Murashige and Skoog (MS) medium. The cultured medium contained 3% sucrose and 0.3% phytagel. There were several combinations levels of 2,4 Dichlorophenoxyacetic acid (2,4-D) and Benzylaminopurine (BAP). The most effective combination treatment for multiple-shoot clumps induction was 0.1 mg L-1 2,4-D plus 2.0 mg L−1 BAP. The addition of 5µM CuSO4 could increase the percentage of clump proliferation. As much as 57% plant regeneration frequency was achieved by culturing the clumps on a solid MS medium containing 0.01 mg L−1 NAA and 2.0 mg L-1 BAP. All registrants were successfully grown up in the soil. Comparison of results from morphological characteristics evaluation and DNA content using flow cytometry analysis showed that in vitro regenerants did not reveal any significant difference compared with control plants. The research findings indicated that the combination between 2,4-D, BAP, and low concentration of CuSO4 improved the multiple shoot clumps formation of Merkeron. [ABSTRACT FROM AUTHOR]

The role of T-Regulatory Cell (Treg) and Interleukin 10 (IL-10) in the failure of BCG protection is unclear. This case-control study analyzed quantity of Treg Cells and IL-10 in the circulation of pediatric TB and non-TB patients with both groups has history of BCG vaccination. Measurement of circulatory Treg cell percentage was performed using Flow Cytometry and circulatory IL-10 was measured using ELISA method. We found that pediatric TB patients have significantly higher IL-10 level compared to non-TB patients (9.15 pg mL−1 vs. 6.04 pg mL−1; p < 0.001). The same scenario happened in Treg cell percentage that were almost two-fold higher in pediatric TB patients compared to non-TB patients (3.51% vs. 1.86%; p < 0.001). It is concluded that the percentage of Treg cells and IL-10 levels in circulation of pediatric TB patients were higher compared to non-TB patients. This finding strengthens the possibility of Treg and IL-10 involvement in the failure of BCG protection. [ABSTRACT FROM AUTHOR]

The effect of the L-ascorbate 1-(2-hydroxyethyl)-4,6-dimethyl-1,2-dihydropyrimidine-2-one on blastema regeneration in Girardia tigrina planarians was investigated. It was established that the preparation has low toxicity towards planarians, which is evidenced by its CL50 values (lethal concentration). The results of the vital computer morphometry and flow cytofluorometry confirm that L-ascorbate 1-(2-hydroxyethyl)-4,6-dimethyl-1,2- dihydropyrimidine-2-one induces regeneration at high concentrations only: 0.01 g/100 mL, 0.05 g/100 mL, and 0.1 mg/100 mL. Low concentrations of the preparation have no influence on blastema regeneration in planarians. [ABSTRACT FROM AUTHOR]

Perspectives on cell therapy using multipotent stem cells or progenitor stem cells are progressing rapidly. Therefore, biomedical research on cells detection method should in line with it. Flow cytometry technique is usually used quantitatively to see the presence of stem cells in different sources, such as blood, bone marrow, tissues, etc. Nevertheless, there is another modality that can be used as an alternative, i.e. polymerase chain reaction (PCR). This study aimed to know whether PCR could be used as an alternative method for replacing flow cytometry technique in detecting human mononuclear cells (hMNCs) derived from umbilical cord blood in the rat blood. Human mononuclear cells from umbilical cord blood were injected into rat tail’s vein. Blood sampling was collected from the heart 30 minutes, 1 hour, 3 hours, 6 hours, 1 day and 7 days after injection. The blood was then analyzed with flow cytometry to determine the presence of hMNCs (CD34+ hMNCs) and PCR to determine the presence of human DNA (human chromosome 17 alpha satellite DNA; HSA17). Polymerase chain reaction could be detecting the presence of human DNA in all samples that are compared to the flow cytometry that also detect the presence of hMNCs in all samples. The results showed that flow cytometry and PCR technique have a similar trend pattern based on time dependent. Polymerase chain reaction techniques can be used as an alternative method in detecting the presence of hMNCs in the rat’s blood equally to the flow cytometry. [ABSTRACT FROM AUTHOR]

Chalcones have been widely researched its activity as anticancer. The substitution of the functional groups on the chalcones structure affects its anticancer activity. The objective of this research were to investigate the cytotoxic effects of bromo chalcone [1-(4'-bromophenyl)-3-(4-hydroxy-3-methoxyphenyl)-2-propene-1-on] or BHM on T47D breast cancer cells by invitro, and to analyze its effect on cell apoptosis. Cytotoxic activity of BHM on T47D cells was assayed by MTT (3-[4, 5-dimethylthiazol-2-yl)-2, 5 diphenyl tetrazolium bromide] method, while the effect of apoptosis was analyzed using flowcytometer. Furthermore, the apoptotic mechanism was traced by observing the expression of Bcl-2 protein using immunocytochemistry. The results indicated that BHM have cytotoxic effect on T47D cells (IC50 = 45 micro molar). The activity occurs because these compounds induce apoptosis by decreasing the expression of Bcl-2. According to the results, BHM could potentially be developed as a new drug candidate for breast cancer therapy. [ABSTRACT FROM AUTHOR]

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Cyanobacterial blooms deteriorate water quality and some species produce cyanotoxins, posing a potential risk to human health. As such, rapid and accurate quantification of cyanobacteria is of growing interest worldwide. In this study, two conventional methods, flow cytometry (FCM) and real-time quantitative PCR (qPCR) were compared in terms of quantification of the total cyanobacteria species in a lab-cultured Microcystis aeruginosa and 10 tropical freshwater samples. Results showed that these two methods have high linearity and strong association (P<0.01), although qPCR has higher sensitivity for low concentrations, while FCM approach has much shorter analysis time for a comparable detection range. The 16S rRNA gene copies/cell ranged from 2.31 to 4.07 in natural freshwater samples. The results showed that the FCM method was preferred for routine cyanobacteria monitoring in the picoplankton range. [ABSTRACT FROM AUTHOR]

Tapak Liman (Elephantopus scaber L) is a traditional medicinal plant containing several active compounds that potentially affecting hematopoietic stem cells, such as epifrieelinol, lupeol, stigmasterol, triacontane-1-ol, dotriacontane-1-ol, lupeol acetate,deoxyelephan-topin, isodeoxyelephantopin, polyphenol luteolin-7, as well as various flavonoids and glucosides. The aim of this study was to elucidate the effect of leaf extract of Tapak Liman on hematopoietic stem cells in mice BALB/c, by observation of the relative number of cells expressing CD4/CD8, CD4/CD62L, and TER119/B220 in the spleen and TER119/B220, TER119/VLA-4 and TER119/CD34 in bone marrow, after being administered leaf extract for 2 weeks. This experiment used 12 female mice, which were divided into three treatment groups, P1= 0.5 g.g bw-1.day-1, P2= 1.0 g.g bw-1.day-1 and P3 = 2.0 g.g bw-1.day-1 Tapak Liman leaf extract as well as a control. The relative numbers of cells expressing surface molecules were analyzed by flowcytometry and quantitative data were tested using one-way ANOVA. The results showed that the leaf extract of Tapak Liman has no significant effect on erythrocyte proliferation; on the other hand, it had a significant effect on both proliferation and differentiation of B lymphocytes (B220+) in bone marrow (p=0.044) and increased the expression of CD4+, CD8+ molecule in B cells (p=0.026) and erythroid cells in spleen and bone marrow, based on the estimation of cells that expressed TER119+VLA-4+, identified as important in the development pathway of erythrocytes. An increased cell percentage of TER11+VLA-4+ occurred for treatment P2, 12% higher than the control. The increased expression of TER119+VLA-4+ was assumed to be due to the iron content in Tapak Liman, which functioned to stimulate the progenitor hematopoietic cells to proliferate and differentiate into a precursor of erythroid cells (TER119 +VLA-4+). There was an increasing number of cells expressing the surface molecules TER119+ and VLA-4+. This indicated that the Tapak Liman leaf extract with a dose of 1.0 g.g bw-1.day-1 could stimulate the proliferation of hematopoietic stem cells into the lymphoid and erythroid pathway, in spleen and bone marrow. [ABSTRACT FROM AUTHOR]

Coronary artery disease (CAD) has emerged as a leading cause of death in Indonesia nowadays. WHO data in 2012 revealed that 37% of the Indonesian population died from this disease. CAD occurs because of endothelial dysfunction in the arteries. Lipoprotein-associated phospholipase A2 (Lp-PLA2), also known as platelet-activating factor acetylhydrolase (PAF-AH), is a phospholipase A2 enzyme, encoded by the PLA2G7 gene. This protein is predicted to be involved in inflammatory phospholipid metabolism so it can be used as a biomarker of CAD in the early phase. Thus, the purpose of this research is to discover the difference in antibody production between wild-type and mutant V279F. The PAF-AH enzyme was isolated from mice lymphocyte cells in order to develop this enzyme as a biomarker of cardiovascular disease. PAF-AH migrates at 55kDa according to SDS-PAGE analysis. Flow cytometry analysis showed that mutant PAF-AH (V279F) is more antigenic than wild-type PAF-AH. The missense mutation of V279F PAF-AH means this enzyme cannot catabolize the acetyl group at the sn-2 position of PAF. [ABSTRACT FROM AUTHOR]

Cell sorting and separation are considered to be very crucial preparatory steps for numerous clinical diagnostics and therapeutics applications in cell biology research arena. Label free cell separation techniques acceptance rate has been increased to multifold by various research groups. Size based cell separation method focuses on the intrinsic properties of the cell which not only avoids clogging issues associated with mechanical and centrifugation filtration methods but also reduces the overall cost for the process. Consequentially flow based cell separation method for continuous flow has attracted the attention of millions. Due to the realization of structures close to particle size in micro dimensions, the microfluidic devices offer precise and rapid particle manipulation which ultimately leads to an extraordinary cell separation results. The proposed microfluidic device is fabricated to separate polystyrene beads of size 1 µm, 5 µm, 10 µm and 20 µm. The actual dimensions of blood corpuscles were kept in mind while deciding the particle size of polystyrene beads which are used as a model particles for study. [ABSTRACT FROM AUTHOR]

Flow cytometry is a benchmark technique used for basic research and clinical diagnosis of various diseases. Despite being a high-throughput technique, it fails in capturing the morphology of cells being analyzed. Imaging flow cytometry is a combination of flow-cytometry and digital microscopy, which offers advantages of both the techniques. In this paper, we report on the development of an indigenous Imaging Flow Cytometer, realized with the combination of Optics, Microfluidics, and High-speed imaging. A custom-made bright-field transmission microscope is used to capture images of cells flowing across the microfluidic device. High-throughput morphological analysis on suspension of yeast cells is presented. [ABSTRACT FROM AUTHOR]

Cell-phone based imaging flow cytometry can be realized by flowing cells through the microfluidic devices, and capturing their images with an optically enhanced camera of the cell-phone. Throughput in flow cytometers is usually enhanced by increasing the flow rate of cells. However, maximum frame rate of camera system limits the achievable flow rate. Beyond this, the images become highly blurred due to motion-smear. We propose to address this issue with coded illumination, which enables recovery of high-fidelity images of cells far beyond their motion-blur limit. This paper presents simulation results of deblurring the synthetically generated cell/bead images under such coded illumination. [ABSTRACT FROM AUTHOR]

In these experiments HeLa cells were maintained in culture in MEM + 10% FCS. For experimental trials, HeLa cells were seeded in 96 well plates at a density of 30 x 103 cells/well and stimulated with varying concentrations of progesterone. To determine calcium dynamics, a Fluo-4 NW calcium assay kit (Molecular Probes) was utilized. Briefly, 100 ml of the dye loading solution was added to each well and allowed to incubate at 37°C for 30 min followed by room temperature for an additional 30 min. Fluorescence was then measured on a flow cytometer using instrument settings appropriate for excitation at 494 nm and emission at 516 nm. Results indicated a 40% increase in calcium mobilization at all concentrations of progesterone. These results are encouraging for further studies into the mechanisms of calcium dynamics in mammalian cells. [ABSTRACT FROM AUTHOR]

Gold nanorods (GNR) in laser-induced thermal therapy can significantly increase light absorption, leading to a local temperature increase and causing irreversible cell damage. One of the key challenges in using GNR as a thermal therapy agent is to deliver a concentration of GNR to generate sufficient heat and cause cell death. In this study, ultrasound and microbubble induced sonoporation is used to enhance intracellular uptake of GNR and improve the therapeutic outcome of laserinduced thermal therapy. Acute myeloid leukemia (AML) cells in suspension (0.6 mL) were treated with ultrasound and microbubbles (USMB) at 1 MHz frequency, 16 microseconds pulse duration, 1 kHz pulse repetition frequency, 1 minute insonation time, varying acoustic pressures (0, 1.26 and 1.73 MPa) and 10 μL Definity microbubble agent with and without GNR (12 nm × 48 nm) at varying concentration (1.0×1010 to 2.5×1011 GNR/mL). The GNR were manufactured through wet chemical synthesis process and measured using Transmission Electron Microscopy (TEM) and Atomic Absorption Spectroscopy (AAS) for size and concentration respectively. Following ultrasound and microbubble treatment, cells were centrifuged to remove excess gold nanorods and treated in suspension with an 810 nm laser (Diomed 60 NIR) at 4 W for 5 minutes. A thermal camera (FLIR Thermovision A40) was positioned to monitor the sample temperature throughout laser treatment and cell viability was assessed using flow cytometry with propidium iodide. Cell viability of 18±2% was achieved with GNR+USMB (1.26 MPa) compared to 72±3% with GNR alone (12 hour incubation) and 99±0.2% with USMB (1.26 MPa) alone. With increasing GNR concentration during ultrasound and microbubble treatment, laser induced sample temperature increased and consequently cell viability decreased. Cell viability decreased from 92±1% at 1.0×1011 GNR/mL to 29±5% at 1.5×1011 GNR/mL concentration with corresponding maximum temperatures of 50°C and 54°C, respectively. The combined treatment of ultrasound-microbubble and gold nanorod laser induced thermal-therapy showed a synergistic enhancement of cell death in vitro. This study shows promise for an enhanced therapeutic effect with the combined treatment in vivo. [ABSTRACT FROM AUTHOR]

Flow cytometry has been widely used for the diagnosis of various hematopoietic diseases. Although there have been advances in the number of markers that can be analyzed simultaneously, the data is still interpreted by manual gating. This is labor-intensive, time-consuming, and subject to human error. We propose a computational model to detect B-lymphocyte neoplasms using flow cytometry data by building healthy and sick profiles. A cell capture rate was defined to measure the fitness of a test subject using a particular profile. By examining the cell capture rate of a test case with all profiles, the disease type can be determined. [ABSTRACT FROM PUBLISHER]

We study parallel algorithms for computing matchings in graphs and apply them to solve population registration problem from bio-imaging data. We have developed several classes of multithreaded algorithms for maximum cardinality matching and achieved good speedups on three shared memory machines on a representative set of large real-world and synthetic graphs. The parallel machines include processors that employ multithreading and cache (Intel Nehalem and AMD Opteron) and massively multithreading and flat memory model (Cray XMT). The bio-imaging application involves registering different cell populations across samples using flow cytometry data. The population registration problem is solved by a generalized edge cover, computed from a weighted matching. We have used this approach to differentiate leukemic cells from healthy ones, and to identify phosphorylation shifts in T cells due to stimulation with an antibody. In current work, we are adapting the concept of consistency used in multiple sequence alignments to the population registration problem for large sample sets. [ABSTRACT FROM PUBLISHER]

This work concerns a program (EFlow) that has been developed to assist experts in clustering flow cytometry multivariate data. EFlow aims to reduce subjectivity but keeping the expert's control over the analysis. In the present paper we will focus on the use of the iterative optimization algorithm. This algorithm is a hill-climbing unsupervised procedure that in general guarantees good local but not global optimization. A comparison between clustering processes achieved by an expert in flow cytometry and by EFlow was performed in a data related to bone marrow cells stained for the surface expression of molecules (B220, IgM and CD23) that characterize the B cell lineage. This data set is considered to present a medium difficult and had 63780 cells with 7 channels or dimensions. Two types of clustering processes were compared, one with four clusters and another with five clusters. The results obtained by EFlow were considered satisfactory by the expert as they were significantly in agreement with his classification. In some clusters, EFlow classified correctly up to 100% of the cells. At the present time few commercial cytometry software offer equivalent clustering analysis. [ABSTRACT FROM PUBLISHER]

Remote monitoring and flow cytometry were lately significantly developed. Ballast water treatment systems (BWTS) are in use on lot of ships today and new systems are still in research projects worldwide as well. So fast development requires control and monitoring of installed systems. International Maritime Organization (IMO) regulations and Ballast Water Convention 2004 include inspection of treated ballast water (BW) if needed. That could initiate lot of legal, beside to operational, problems in maritime industry. That implicates the need of a monitoring treated BW before the ship enters a port. It has been described in this article how to monitor treated ballast water to make sure the BW and BWTS on a ship is in proper condition, but before the port entrance. The monitoring system and its parts flow cytometer and satellite communication system together with remote operation system from the land office have been explained. Auto-monitoring of treated BW on board by using newest sensor technologies and satellite communications are included in the model of three sub-systems and remote operation from the land office, but to be autonomous from the ship's crew. Main goal of the research is that a ship enters the port with the BW cleanliness confirmation. In this way there is no need to sample the BW and no time is lost for analysing the samples. [ABSTRACT FROM PUBLISHER]

Experiments were performed to size, count, and obtain shell parameters for individual ultrasound contrast microbubbles using a modified flow cytometer. Light scattering was modeled with Mie theory and applied to calibration beads to calibrate the system. The size distribution and population were measured directly from the flow cytometer. The shell parameters (shear modulus and shear viscosity) were quantified at different acoustic pressures by fitting microbubble response data to a bubble dynamics model. The size distribution of the contrast agent microbubbles is consistent with manufacturer specifications. The shell shear viscosity increases with increasing equilibrium microbubble size, and decreases with increasing shear rate. The observed trends are independent of driving pressure amplitude. The shell elasticity does not vary with microbubble size. The results suggest that a modified flow cytometer can be an effective tool to characterize the physical properties of microbubbles, including size distribution, population, and shell parameters. [ABSTRACT FROM PUBLISHER]

The paper will focus on the discussion of recent unique examples of Finite-Difference Time-Domain (FDTD) simulations and modeling of biophotonics applications including Optical Phase Contrast Microscope (OPCM) imaging of cells containing gold nanoparticles (NPs). The discussion includes the case of OPCM microscopy as a prospective modality for in vivo flow cytometry. The application of the FDTD approach for the simulation of OPCM and flow cytometry imaging opens a new application area with a significant research potential. [ABSTRACT FROM AUTHOR]

Cancer stem cells display extensive differentiation potential and can give rise to new tumours. Therefore, an important attention must be paid for cancer stem cell identification, characterization and investigation of for novel targeted therapies. Quantum dots are promising tools in fighting against cancer. However, there is not much known about accumulation and cytotoxicity of quantum dots in cancer stem cells. It is important to know that before we use quantum dots as therapy agents. [ABSTRACT FROM AUTHOR]

This work investigates the mechanism of sonoporation through the effect of ultrasound exposure parameters with microbubbles in a suspension of KHT-C cells. We investigated the effect of acoustic pressure, pulse repetition frequency, pulse duration, insonation time and frequency. Flow cytometry and fluorescence microscopy were used to quantify permeability and viability of treated cells. Scanning electron microscopy images showed pores of up to 500nm after treatment, indicating that the mechanism of permeabilisation was the formation of pores on the surface of cell membranes. Cell permeability increases and viability decreases with peak negative pressure, duty cycle and insonation time. Higher pulse centre frequencies are more effective at permeabilising cells, but also at killing them. In this study, the highest Therapeutic Ratio is achieved at 570kPa peak negative pressure, 8μs pulse duration, 3kHz pulse repetition frequency, 500kHz centre frequency and 12 seconds of insonation time; Definity at 3.5% volume concentration. This implies that ultrasound can be optimized with respect to the therapeutic application. © 2007 American Institute of Physics [ABSTRACT FROM AUTHOR]

Bioactive substances such as peptides and nucleic acid based agents have attracted great attention for the next generation drug for various diseases. However, the greatest challenge for using these bioactive substances is the development of their delivery system, especially the method for delivering these substances through the cell membrane. With the advancement of ultrasound and ultrasound contrast agent technology, it has become possible to transiently change the permeability of the cell membrane. Moreover, using a focused ultrasound transducer, it is possible to narrow and focus the ultrasound energy within a small target, avoiding damage to the surrounding tissue. In this research we have searched the possibility of delivering the Bak BH3 peptide, the death domain of the Bc1-2 family of proteins, or the short interfering RNA (siRNA) targeting the enhanced green fluorescent protein (EGFP) using microbubble-enhanced focused ultrasound in an in vitro setting. Using a 1.696 MHz focused ultrasound and a microbubble ultrasound contrast agent OPTISON®, we first tested the stability of BH3 peptide under microbubble-enhanced focused ultrasound exposure and proved that the peptide is stable under these circumstances. Next, we have tested the cell-killing effect of the intracellularly delivered Bak BH3 peptide in HeLa and BJAB cell line and observed a statistically enhanced cell death in BJAB cells but not in HeLa cells, leading to the conclusion that intracellularly delivered BH3 peptide by microbubble-enhanced ultrasound can exert its cell killing effect in some cells. We also investigated if we can silence the EGFP expression in the cell by delivering siRNA targeting the EGFP in both transient and stable EGFP expression cell line. Using a 1.653 MHz focused ultrasound and OPTISON®, in both cases, intracellularly delivered siRNA by microbubble-enhanced ultrasound was able to knock down the EGFP expression, which demonstrates the feasibility of using this novel method for siRNA intracellular delivery. In conclusion, our investigation has proved the feasibility of using micro-bubble enhanced focused ultrasound as a method for intracellular delivery of peptides and siRNAs. Although further optimization of various parameters is necessary, we consider that this method could be a power tool for using these bioactive substances in the clinical field. © 2006 American Institute of Physics [ABSTRACT FROM AUTHOR]

This paper will examine the debate surrounding the use of flow cytometry and preimplantation genetic diagnosis to sex select children for social reasons in order to interrogate the meanings of sexed sperm and embryos. By analyzing this specific node within the larger domain of both sex selection and the new reproductive technologies, I hope to create an initial site of entry into the complex webs that have constructed this controversy. A sample of written publications from May 2001 ? May 2002, appearing in newspapers, popular magazines and science/medical/bioethics journals, have been examined. This paper will begin by mapping the social worlds that have represented their positions within these media on preconception sex selective technologies; specific focus will be paid to the meanings that sorted sperm and sexed embryos have to each group. I hope to further problematize the ethical debate surrounding sex selection by considering whose voices were not articulated in the resolution of this debate in order to highlight the role of power. The paper will conclude with a discussion of how divergent groups came together to translate the diverse meanings of sex selected sperm and embryos during a key historical moment in both the technical and sociocultural trajectories in order to maintain the project of sex selecting children. [ABSTRACT FROM AUTHOR]

The genus Iva contains 9 species of semi-woody shrubs and herbs. This genus is centered in North America and the West Indies. We collected and used DNA flow cytometry to generate C-values for three Iva species. The C- value is a measure of the amount of DNA in an organism's nuclear genome. We used the C-values to compare the genome sizes for each of the Iva species included in the study, which we hope to relate to ploidy level variation to explore speciation and evolution in the genus. Baseline C-value data in this genus is required to explore genome evolution in a lost domesticated crop, sumpweed (Iva annua). The seeds of Iva annua, sumpweed, have been posited as a pre-maize food crop of indigenous Americans in the mid-western and eastern regions of the USA. The genome variation if detected in extant populations of Iva annua may represent vestiges of phenotypes related to historical domestication. [ABSTRACT FROM AUTHOR]

I will present a microfluidic imaging flow cytometer incorporating stroboscopic illumination, for blur-free cellular analysis at throughputs exceeding 100,000 cells per second. By combining passive (inertial or viscoelastic) focusing of cells in parallel microchannels with stroboscopic illumination, such chip-based cytometers are able to extract multi-colour fluorescence and bright-field images of single cells moving at high linear velocities. This in turn allows accurate sizing of individual cells, intracellular localization and analysis of heterogeneous cell suspensions. The method is showcased through the rapid enumeration of apoptotic cells, high-throughput discrimination cell cycle phases and localization of p-bodies. [ABSTRACT FROM AUTHOR]

Previously, we reported that macrophage (H-2DbKb)-mediated cytotoxicity of allografts [e.g., BALB/c skin (H-2DdKd)] in the rejection site and that we isolated two cDNAs encoding novel receptors on mouse macrophages or monocytes for H-2Dd and H-2Kd (mouse MHCs) and named them MMR1 and 2. Recently, we identified human homologs of mouse MMR1 and 2 by a search of sequence homology (BLAST) and also isolated two cDNAs encoding human MMR1 and 2 from human PBMCs cDNA library by RT-PCR method. In this meeting, we present the identification and characterization of human MMR1. The predicted amino acid sequence of human MMR1 cDNA (approximately 1.5 kb) encoded a putative single transmembrane protein, and the homology showed 64% identity to the entire mouse MMR1 protein and 96%, 96%, and 98% identity to its respective intracellular, transmembrane, and MHC binding regions. RT-PCR analyses demonstrated that human MMR1 expressed on monocytes in PBMCs. To examine the binding toward HLAs (human MHCs), HEK293T cells expressing human MMR1 were incubated with the beads conjugating 80 kinds of HLA class I or fluorescein-labeled soluble HLA molecules and then were analyzed by flow cytometer. Flow cytometric analyses showed that the mean fluorescence intensity of human MMR1 protein toward HLA-B44 was significantly high and that HEK293T cells expressing human MMR 1 bound specifically to HLA-B44 with a dissociation constant of 3.0 x 10-9 M. These results suggest that human MMR 1 on monocytes or macrophages might be novel receptors for HLA-B44. [ABSTRACT FROM AUTHOR]