Coulter Counter Research Articles

Distribution and risk factors of Trichomonas vaginalis infection with relevance to interleukin profile among population in Duhok city, Iraq

BackgroundVaginal trichomoniasis is a sexually transmitted disease causes adverse health outcomes, the causative agent is Tichomonas vaginalis. Infected women develop abnormal vaginal discharge with vaginitis. While, men mostly suffer from prostatitis and urethritis. ObjectivesThis study aimed to detect the rate of T.vaginalis infection in association with risk factors, measurement of blood parameters, interleukine-2 (IL-2) and interleukine-8(IL-8) levels among individuals. MethodsA total of 500 females were tested, a vaginal swab and blood were collected from the participants, and they were within the age range of 18–70 years old. High vaginal swabs were examined microscopically using wet mount preparation, the blood used for measurement of blood parameters using coulter count machine and IL-2 and IL-8 levels were evaluated using ELISA technique. ResultsOut of 500 individuals, the prevalence of T. vaginalis infection was 45(9.0 %). The maximum infection rates were reported among age group (18–28) years, rural residents, illiterates, married women (16.7 %), (15.9 %),(30.0 %),(11.7 %),respectively. Moreover, high positivity of infection revealed among low economical level 12.5 % compared to only 1.7 % in high economy group. Unemployed women, those with bad vaginal hygiene showed high incidence of infection 10.5 % and 6.3 %, respectively. It was noted a significant decrease in red blood cells and hemoglobin concentration, however, there was a significantly increased in white blood cells among infected subjects. Furthermore, the levels of both interleukin-2 and interleukin-8 were significantly higher among infected individuals compared to healthy ones. ConclusionsEffective strategies for disease awareness are needed and should include health promotion, education, and prevention.

Simultaneous enumeration of yeast and bacterial cells in the context of industrial bioprocesses

Abstract In scenarios where yeast and bacterial cells coexist, it is of interest to simultaneously quantify the concentrations of both cell types, since traditional methods used to determine these concentrations individually take more time and resources. Here, we compared different methods for quantifying the fuel ethanol Saccharomyces cerevisiae PE-2 yeast strain and cells from the probiotic Lactiplantibacillus plantarum strain in microbial suspensions. Individual suspensions were prepared, mixed in 1:1 or 100:1 yeast-to-bacteria ratios, covering the range typically encountered in sugarcane biorefineries, and analyzed using bright field microscopy, manual and automatic Spread-plate and Drop-plate counting, flow cytometry (at 1:1 and 100:1 ratios), and a Coulter counter (at 1:1 and 100:1 ratios). We observed that for yeast cell counts in the mixture (1:1 and 100:1 ratios), flow cytometry, the Coulter counter, and both Spread-plate options (manual and automatic CFU counting) yielded statistically similar results, while the Drop-plate and microscopy-based methods gave statistically different results. For bacterial cell quantification, the microscopy-based method, Drop-plate, and both Spread-plate plating options and flow cytometry (1:1 ratio) produced no significantly different results (P > 0.05). In contrast, the Coulter counter (1:1 ratio) and flow cytometry (100:1 ratio) presented results statistically different (P < 0.05). Additionally, quantifying bacterial cells in a mixed suspension at a 100:1 ratio wasn't possible due to an overlap between yeast cell debris and bacterial cells. We conclude that each method has limitations, advantages, and disadvantages.

Photothermal Enhancement of Ion Current for Red Blood Cell Flow Cytometry.

Blood cell counting typically requires complex machinery. Flow cytometers used for this purpose involve precise optical alignment, costly detectors, and pretreatment with fluorescent labels. Coulter countertype devices, which monitor ion current, are simpler. However, conventional Coulter counters provide only information about size, making it impossible to distinguish similarly sized lymphocytes from red blood cells (RBCs). Inspired by the fact that RBCs have an exceptionally high propensity to absorb light and convert it to heat, i.e., photothermal effect, this study proposes integrating photothermal phenomena into a microfluidic Coulter counting chip. Photothermal heat selectively amplifies the ion current from RBCs over other components including lymphocytes. The combination of ion current monitoring and the photothermal effect for RBC counting suggests an evolution toward versatile flow cytometers.

High Surface Area Metal Oxide Enhanced Self-Powered Microfluidic Device for Real-Time Pathological Detection

Biomarkers based sensing in the realm of disease prevention, its diagnosis, and drug development has shown its unique importance in health care. In particular, the platelet level in blood is strongly linked with the development of several kind of severe diseases like coronavirus disease 2019, diabetes, and microbial infections as well as individuals undergoing cancer diagnosis. Currently, commercial methods employed for platelet level determination in whole-blood are based on Coulter principle, flow cytometry, and image analysis. However, these technologies are intricate, time consuming, expensive, and require a trained operator as well as lack of portability. Considering these issues, we have developed a self-powered portable device for real-time platelet level monitoring, aimed towards point-of-care applications. The fabricated self-powered-microfluidic triboelectric nanogenerator (SPM-TENG) utilized test sample flow-resistance for sensing of platelet level and triboelectric voltage output as sensing signal. For the fine tuning of fabricated SPM-TENG, we compared the triboelectric voltage output from gold electrode measurement setup and terminal electrode measurement setup. In view of further improvement in stability, voltage output and to minimize the noise in sensing signals, the geometrical optimization of microfluidic channel was performed. In conclusion we have optimized the device as a terminal electrode measurement setup with copper tubes on terminals of microfluidic channel and copper oxide nanowires grown on the interior of copper tubes. These copper oxide nanowires provide a high surface area for solid-liquid contact separation and is responsible for enhanced sensing signal. The fabricated SPM-TENG has shown its great potential in platelet level sensing with plasma-based samples as well as whole-blood samples. The proposed device demonstrated its practicality towards real-time rapid sensing of platelet level and portability.

Osmotic injury and cytotoxicity for hMSCs in contact with Me2SO: The effect of cell size distribution

The paper discusses the impact of cell size on cytotoxicity and expansion lysis during the osmotic excursions resulting from the contact of hMSCs from UCB with Me2SO. It builds upon the mathematical model recently presented by the authors, which pertains to a population of cells with uniform size. The objective is to enhance the model's relevance by incorporating the more realistic scenario of cell size distribution, utilizing a Population Balance Equations approach. The study compares the capability of the multiple-sized model to the single-sized one to describe system behavior experimentally measured through cytofluorimetry and Coulter counter when, first, suspending hMSCs in hypertonic solutions of Me2SO (at varying osmolality, system temperature, and contact times), and then (at room temperature) pelleting by centrifugation before suspending the cells back to isotonic conditions. Simulations demonstrate that expansion lysis and cytotoxic effect are not affected by cell size for the specific system hMSCs/Me2SO, thus confirming what was found so far by the authors through a single-size model. On the other hand, simulations show that, when varying the adjustable parameters of the model that are expected to change from cell to cell lineages, expansion lysis is sensitive to cell size, while cytotoxicity is not, being mainly influenced by external CPA concentration and contact duration. More specifically, it is found that smaller cells suffer expansion lysis more than larger ones. The findings suggest that different cells from hMSCs may require a multiple-sized model to assess cell damage during osmotic excursions in cryopreservation.

Quantitative signs on ultrasound biomicroscopy images associated with the status of silicone oil emulsification within the eye.

Silicone oil (SO) is a commonly used intraocular tamponade in the treatment of rhegmatogenous retinal detachment (RRD). SO emulsification is a common complication after SO injection. This study aimed to investigate correlations between SO emulsification signs on ultrasound biomicroscopy (UBM) and the real number/size of SO droplets. An observational, cross-sectional study. Thirty-five RRD patients (19 males; 35 eyes) who underwent pars plana vitrectomy and SO injection were enrolled in the study. SO emulsification signs on UBM (endothelial deposits, floating droplets, ghost images, hyperoleon, anterior chamber angle impregnation, anterior iris surface impregnation, posterior iris surface impregnation and ciliary body impregnation) were acquired and quantified two days before SO removal, while real number/size of the SO droplets with a diameter in 0.4-20 μm in the first 2 mL washout fluid collected intraoperatively during SO removal were assessed by Coulter counter. Then the correlations between these values were explored. The grade of tissue impregnation and hyperoleon area in the anterior chamber demonstrated significant correlations with the number of large (5-20 μm) SO droplets (r = 0.336; P = 0.048 and r = 0.350; P = 0.039, respectively); however, no significant correlations were observed with the total number of droplets (0.4-20 μm)or the number of small (0.4-5 μm) droplets (both P ＞ 0.05). The UBM quantitative grade of tissue impregnation and hyperoleon area in the anterior chamber could provide a noninvasive overview of the actual degree of SO emulsification before SO removal surgery.

A Study of the Mean Platelet Volume and Plasma Fibrinogen in Type Two Diabetes Mellitus Patients Versus Healthy Controls and Their Role as Early Markers of Diabetic Microvascular Complications.

Background Mean platelet volume (MPV) is considered an emerging biological marker of platelet function and activity. Higher MPV has been scientifically linked to diabetes mellitus, metabolic syndrome, stroke, and coronary artery disease. Plasma fibrinogen is a circulating glycoprotein, serving as an acute inflammatory marker ultimately leading to enhanced atherogenic plaque formation. We conducted this study to evaluate the crucial role of MPV and plasma fibrinogen, which showed elevated levels in diabetes mellitus patients compared to non-diabetic healthy individuals. This study also elaborates on the pivotal role that MPV and plasma fibrinogen levels play in the pathogenesis of microvascular complications, which progress and eventually lead to mortality in patients with type 2 diabetes mellitus. Methodology This study is a single-center hospital-based study including120 type 2 diabetes mellitus patients and 120 healthy non-diabetic individuals. It is across-sectional and observational study. The study wasconductedover a period of one and a half years in a medical college and hospital in a semi-urban locality in Western Maharashtra, India. We obtained informed written consent from the patients. All patients underwent thorough clinical assessment, and data were collectedusing proformas, which were latertabulated and entered in Microsoft Excel sheets. Later, the statistical data analysis was performed.Plasma fibrinogen was performed by photo-optical clot detection. MPV was analyzed by coulter principle in the central laboratory department of the parent institute. Patients above 18 years with cases of type 2 diabetes mellitus with or without any related complications, while the controls are healthy non-diabetic individuals attending the outpatient and inpatient departments of General Medicine. We excluded patients under the age of 18 years, those diagnosed with type 1 diabetes mellitus, hematological conditions associated with anemia and abnormal platelet counts, pregnant females, any acute or chronic infections, patients currently on antiplatelet medication and other drugs affecting the platelets, and all critical patients. Results The majority of patients in our study were in the age group of 41-50 years, with 49.2% having one or more microvascular complications of diabetes mellitus. In our study, out of 120 cases, 3.3% and 23.3% had raised MPV and fibrinogen levels, respectively, above the normal range. When compared with males and females, there was no statistically significant difference in the mean value of MPV and fibrinogen. On the t-test (p < 0.05), there was a statistically significant difference in the mean value of MPV and fibrinogen level between diabetics with and without microvascular complications. The t-test (p < 0.05) showed that there was a statistically significant difference among cases in the mean values of MPV and plasma fibrinogen in relation to retinopathy, nephropathy, and neuropathy, which are all microvascular complications of diabetes. Conclusion The study reveals higher levels of MPV and fibrinogen in diabetic patients compared to non-diabetic healthy individuals. In addition,higher levels of MPV and fibrinogen were present in patients with microvascular complications, correlated with age and diabetes duration.

Reconciling Coulter Counter and laser diffraction particle size analysis for aquaculture monitoring

The disaggregated inorganic grain size (DIGS) of bottom sediment analyzed with a Coulter Counter (CC) has been used to show that the fraction of sediment deposited in flocs (floc fraction) increased in both the near and far field after the introduction of open cage salmon aquaculture, altering benthic habitat and species composition. As a result, DIGS was identified as a potential indicator of regional environmental changes due to aquaculture. Laser diffraction is an attractive alternative to the CC because of its greater efficiency and larger size range. To determine if a laser diffraction instrument, Beckman-Coulter LS 13 320 (LS), could replace the CC within a Canadian national aquaculture monitoring program, the DIGS of 581 samples from five different regions in eastern Canada were analyzed with an LS and a CC. Results show that the LS could not be used to calculate floc fraction. Instead, % sortable silt and the volume % of inorganic particles < 16 µm were evaluated as alternative proxies for fine sediment properties. LS and CC values for these parameters were correlated, but they were significantly different and the relationship between the instruments was dependent on the area sampled. The LS did not capture variations between areas seen in the CC DIGS data. Where the DIGS from the CC found no sorting in the finest size classes, all the LS samples had similar size distributions characterized by smooth modal peaks. The LS and CC both return values that can be used to monitor changes in the deposition of fine-grained sediment, but the LS cannot determine changes in floc deposition and caution is required if comparing different sedimentary environments.

An Investigation of Size Distribution and Calcium Signaling in Human Platelets.

Background Platelets are thin disc-shaped blood cells that play a major role in hemostasis, maintenance of vascular integrity, and blood coagulation. Large platelets are more reactive and seen in patients with cardiovascular disease. This study aims to analyze the changes in platelet size of ex vivo activated platelets which phenotypically simulates that of a patient at risk of cardiovascular disease and elucidate the calcium signaling pathway responsible for this change. Methodology Platelets were isolated from adult human blood by differential centrifugation. Calcium was mobilized into platelets by treatment with calcium ionophore A23187 in the presence of Ca2+. Platelet size distribution was analyzed using Coulter Counter Multisizer 4. The following signaling parameters were studied: intracellular Ca2+ measurement (using Fura-2/AM by fluorescence spectrophotometry), Ca2+-dependent thiol protease calpain assay (using fluorogenic substrate t-butoxycarbonyl-Leu-metchloromethylcoumarin in fluorescence microplate reader), platelet-derived microparticles (using FACS Calibur flow cytometry), and cytoskeletal protein talin expression (by western immunoblotting). Results When adult platelets were treated with A23187 and Ca2+, two subcellular populations (<2 µm and between 2-4 µm) were noted. The mean size of the second cell population was significantly higher than that of resting platelets (2.94 ± 0.13 µm vs. 2.82 ± 0.15 µm, t = 4.605, p = 0.00). A23187 treatment led to elevated intracellular Ca2+, release of platelet-derived microparticles, increase in calpain activity, and cytoskeletal talin degradation. These events were inhibited by calpeptin (a specific calpain inhibitor). Conclusions Elevated calcium caused talin degradation by calpain activity. Breakdown of this cytoskeletal protein leads to relative swelling of cells reflected by the increase in platelet size.

Growth and Cell Size of Microalga Auxenochlorella protothecoides AS-1 under Different Trophic Modes.

Certain microalgal species can grow with different trophic strategies depending on the availability of nutrient resources. They can use the energy from light or an organic substrate, or both, and can therefore be called autotrophs, heterotrophs, or mixotrophs. We recently isolated a microalgal strain from the microplastic biofilm, which was identified as Auxenochlorella protothecoides, AS-1. Strain AS-1 grew rapidly in bacterial culture media and exhibited different growth rates and cell sizes under different trophic conditions. We compared the growth performance of AS-1 under the three different trophic modes. AS-1 reached a high biomass (>4 g/L) in 6 days under mixotrophic growth conditions with a few organic carbons as a substrate. In contrast, poor autotrophic growth was observed for AS-1. Different cell sizes, including daughter and mother cells, were observed under the different growth modes. We applied a Coulter Counter to measure the size distribution patterns of AS-1 under different trophic modes. We showed that the cell size distribution of AS-1 was affected by different growth modes. Compared to the auto-, hetero- and mixotrophic modes, AS-1 achieved higher biomass productivity by increasing cell number and cell size in the presence of organic substrate. The mechanisms and advantages of having more mother cells with organic substrates are still unclear and warrant further investigations. The work here provides the growth information of a newly isolated A. protothecoides AS-1 which will be beneficial to future downstream applications.

In Vitro and In Vivo Behavioral Evaluation of Condensed Lipid-Coated Perfluorocarbon Nanodroplets

BackgroundPhase-shift contrast agents consist of a liquid perfluorocarbon core that can be vaporized by ultrasound to generate echogenic contrast with excellent spatiotemporal control. The purpose of the present work was to evaluate the in vitro and in vivo behavior of condensed lipid-shelled nanodroplets using different analytical procedures. MethodsIn this study, perfluorobutane nanodroplets were prepared by condensation of precursor fluorescently labeled lipid shelled microbubbles and were characterized in terms of size distribution, gas core content, in vitro stability in blood as well as for their acoustic vaporization behavior using a custom-made set-up. In particular, their in vivo behavior was thoroughly investigated after intravenous bolus injection in rats. To this end, we report, for the first time, the efficient use of three complementary detection procedures to assess the NDs in vivo persistence namely 1) Ultrasound contrast imaging of vaporized NDs, 2) Gas chromatography-mass spectrometry to determine the perfluorobutane core content and 3) Fluorescence intensity measurement in the collected blood samples. ResultsThe Coulter Counter Multisizer results confirmed the size distribution shift post-condensation. Furthermore, similar PFB concentration from MBs and NDs suspensions was obtained, thus demonstrating an exceptionally low rate of microbubbles breaking and spontaneous nanodroplets vaporization. As expected, these nanoscale droplets display longer circulation time compared to clinically approved microbubbles and only slight variations of half-life were observed between the three monitoring procedures. Finally, echogenic signal observed in focal area of the liver and spleen after vaporization was confirmed by accumulation of fluorescent nanodroplets in these organs. ConclusionThese results further contribute to deepen our understanding of both in vitro and in vivo behavior of sonoresponsive nanodroplets, which is key to enable efficient clinical translation.

Hematological and Biochemical Profiles of Patients with Dengue Virus Infection in Lalitpur, Nepal

Dengue viral infection is a disease transmitted by mosquitoes of the Aedes genus, and it arises as a result of being bitten by mosquitoes that are carrying the virus. This infection induces a wide range of changes in the body’s immune system. Thus serological evaluation together with hematological and biochemical parameters in cases of dengue viral infection make an accurate clinical diagnosis. The main objective of this study was to study the serological, hematological, and biochemical parameters among patients infected with Dengue virus. A total of 102 blood samples were collected at Asrik Diagnostic Lab, Lalitpur during the period of August to October 2022. Samples were analyzed by following methods: hematological tests were done by coulter counter, Biochemical tests were done by fully automated analyzer and serological tests were done by ELISA. Of the 102 blood samples NS1 antigen was detected in 62 (60.9%) whereas anti-dengue IgM antibody was detected in 10 (9.8%). Leukopenia and thrombocytopenia were present in 33 (53.1%) and 32 (51.6%) of the 62 NS1 positive samples, respectively. Of the 62 cases, only 16 samples were tested for the alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Out of 16 samples, ALT and AST level were significantly higher in 11 (68.8%) and 14 (87.5%) positive cases, respectively. The statistical analysis showed that there was no significant association between NS1 and IgM. It can be concluded that the investigation of leukocytes, thrombocytes, ALT, and AST helps to diagnose the dengue efficiently.

Oxidative stress and pro-inflammatory cytokines following perchloroethylene exposure in young female dry-cleaning workers.

The involvement of Perchloroethylene (PCE) in the development of autoimmune diseases has been reported. However, few studies investigated immunotoxicity in PCE-exposed workers. To study changes in the oxidative stress and cytokine profile of young female dry-cleaning workers exposed to PCE. Thirty-eight exposed workers and 38 unexposed controls were recruited. All the participants were young nonsmoker females. Individual interviews were conducted by a physician. Blood samples were collected and hematological tests were performed by an automated Coulter Counter. Plasma PCE levels were determined using gas chromatography/flame ionization detection. Plasma total antioxidant capacity (TAC), Catalase (CAT), Superoxide dismutase (SOD), and Malondialdehyde (MDA) levels were measured using the colorimetric method. The levels of plasma cytokines interleukin-1β (IL-1β), IL-2, IL-4, IL-6, IL-8, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) were measured by commercially kits. The levels of plasma PCE averaged 561±96 ng/ml in the exposed group compared with 1.3±0.5 ng/ml in the controls. The hematological tests failed to find abnormalities in the exposed workers. Exposed workers presented significantly increased plasma levels of MDA, SOD and CAT. There were no significant differences between the two groups for level of plasma TAC. Significantly increased plasma IL-1β and TNF-α and decreased IL-2 and IL-8 levels were seen in the exposed workers. There were no significant differences between the two groups for IL-4, IL-6, and IFN-γ. PCE exposure resulted in changed cytokine profile in dry-cleaning workers, suggesting the potential immunotoxicity of PCE at low exposure levels.

A Wash-Free Spheres-on-Sphere Strategy for On-Site and Multiplexed Biosensing.

Respiratory infections and food contaminants pose severe challenges to global health and the economy. A rapid on-site platform for the simultaneous detection of multiple pathogens is crucial for accurate diagnosis, appropriate treatment, and a reduced healthcare burden. Herein, we present a spheres-on-sphere (SOS) platform for multiplexed detection using a portable Coulter counter, which employs millimeter- and micron-sized spheres coupled with antibodies as multitarget probes. The assay allows for quantitative detection of multiple analytes within 20 min by simple mixing, enabling on-site detection. The platform shows high accuracy in identifying three respiratory viruses (SARS-CoV-2, influenza A virus, and parainfluenza virus) from throat swab samples, with LOD of 50.7, 32.4, and 49.1 pg/mL. It also demonstrates excellent performance in quantifying three mycotoxins (aflatoxin B1, deoxynivalenol, and ochratoxin A) from food samples. The SOS platform offers a rapid on-site approach with high sensitivity and specificity for applications in resource-limited settings.

Streamlined measurement of chimeric antigen receptor T-cell concentration, size, viability and two-color phenotyping during manufacturing

Background aimsThe successful development of CD19-targeted chimeric antigen receptor (CAR) T-cell therapies has led to an exponential increase in the number of patients recieving treatment and the advancement of novel CAR T products. Therefore, there is a strong need to develop streamlined platforms that allow rapid, cost-effective, and accurate measurement of the key characteristics of CAR T cells during manufacturing (i.e., cell number, cell size, viability, and basic phenotype). MethodsIn this study, we compared the novel benchtop cell analyzer Moxi GO II (ORFLO Technologies), which enables simultaneous evaluation of all the aforementioned parameters, with current gold standards in the field: the Multisizer Coulter Counter (cell counter) and the BD LSRFortessa (flow cytometer). ResultsOur results demonstrated that the Moxi GO II can accurately measure cell number and cell size (i.e., cell volume) while simultaneously assessing simple two-color flow cytometry parameters, such as CAR T-cell viability and CD4 or CAR expression. ConclusionsThese measurements are comparable with those of gold standard instruments, demonstrating that the Moxi GO II is a promising platform for quickly monitoring CAR T-cell growth and phenotype in research-grade and clinical samples.

Initial investigations into the production of triploid sunshine bass using temperature shock

AbstractObjectiveThe sunshine bass (female White Bass Morone chrysops × male Striped Bass M. saxatilis) is an important food fish raised in U.S. aquaculture. Spawning is performed by manually stripping and fertilizing White Bass eggs with Striped Bass sperm. Embryos hatch in approximately 48 h. Mature sunshine bass females can become fertile and produce viable eggs, which is problematic in commercial food‐fish production; therefore, triploid fish are desired.MethodsThe present nonreplicated study incorporated three warm and three cold temperature shocks, along with three postfertilization initiation times and three exposure times for each temperature, during initial stages of incubation to induce triploidy. After exposure to temperature shocks, fertilized eggs were incubated in experimental hatching systems. The best performing cold and warm temperatures and times (and an ambient diploid control) were then used to incubate fertilized eggs in a commercial‐scale trial using standard industry practices with a single batch of eggs. Larvae were stocked at about 185,000 larvae/ha (75,000 larvae/acre) in newly filled and fertilized ponds. Fry were sampled weekly and harvested at 30 days.ResultThere were 15 survivors from the pond containing fish derived from cold‐shocked fertilized eggs; these individuals were tested via Coulter Counter, and no triploids were detected. Testing of 50 survivors from the pond containing fish derived from warm‐shocked fertilized eggs indicated that 14% were triploid.ConclusionTherefore, initial trials indicate that warm temperature shock is preferable to cold shock for inducing a greater percentage of triploid sunshine bass.

Multiplexed electrical detection of whole viruses from plasma in a microfluidic platform.

The advancement of point-of-care diagnostics is crucial to improving patient outcomes, especially in areas with low access to hospitals or specialized laboratories. In particular, rapid, sensitive, and multiplexed detection of disease biomarkers has great potential to achieve accurate diagnosis and inform high quality care for patients. Our Coulter counting and immunocapture based detection system has previously shown its broad applicability in the detection of cells, proteins, and nucleic acids. This paper expands the capability of the platform by demonstrating multiplexed detection of whole-virus particles using electrically distinguishable hydrogel beads by demonstrating the capability of our platform to achieve simultaneous detection at clinically relevant concentrations of hepatitis A virus (>2 × 103 IU mL-1) and human parvovirus B19 virus like particles (>106 IU mL-1) from plasma samples. The expanded versatility of the differential electrical counting platform allows for more robust and diverse testing capabilities.

Comparing the incidence of spontaneous autopolyploidy in wild and hatchery Lake Sturgeon

AbstractObjectiveLiving species of Acipenseriformes, sturgeons and paddlefishes, are characteristically polyploid, having more than two complete sets of chromosomes (&gt;2n). They undergo spontaneous autopolyploidy, an unintentional one and a half times increase in genome size, more frequently than any other order of fish. For Lake Sturgeon Acipenser fulvescens, which are evolutionary octoploids (8n), spontaneous autopolyploidy results in fertile dodecaploid (12n) progeny. When 12n individuals reproduce with octoploids, it is possible that resulting decaploid (10n) offspring will have poor physiological performance and survivorship. Spontaneous autopolyploidy in the wild is very low; however, incidence in fish hatcheries is greater, as seen in other 8n sturgeon species. We investigated this disparity in Lake Sturgeon, predicting to find more dodecaploid individuals in hatchery populations than in the wild.MethodsPloidy was determined using red blood cells from individuals in three hatchery and two wild populations of Lake Sturgeon in Manitoba, Canada (n = 1004). Red blood cell volume was evaluated with a Z2 Coulter counter and used to determine ploidy, based on the average of triplicate measures of the erythrocyte modal nuclei volume (fL). A subsample from each environment type was further examined using blood smear analysis (n = 130) and flow cytometry (n = 27).ResultOne 12n hatchery individual was found, along with significant differences in erythrocyte morphometry between the five populations. Fluctuations in modal nuclei volume were also observed over 169 days of repeated measurement within a single hatchery population.ConclusionThe well‐developed relationship between erythrocyte size and fishes external and physiological environment may explain the variance both between and within populations. These results demonstrate the need for ploidy monitoring in artificial hatcheries, as releasing even a single 12n fish could produce thousands of 10n offspring that, if recruited, would have a detrimental effect on the population fitness.

New erythrocyte parameters derived from the Coulter principle relate with red blood cell properties-A pilot study in diabetes mellitus.

In routine hematological instruments, blood cells are counted and sized by monitoring the impedance signals induced during their passage through a Coulter orifice. However, only signals associated with centered paths in the aperture are considered for analysis, while the rejected measurements, caused by near-wall trajectories, can provide additional information on red blood cells (RBC), as recent publications suggest. To assess usefulness of two new parameters in describing alterations in RBC properties, we performed a pilot study to compare blood samples from patients with diabetes mellitus (DM), frequent pathological condition associated with impairment in RBC deformability, versus controls. A total of 345 blood samples were analyzed: 225 in the DM group and 120 in the control group. A diagram of [Formula: see text] and [Formula: see text], the two new parameters derived from the analysis of impedancemetry pulses, was used to compare distribution of RBC subpopulations between groups. To discriminate RBC from DM and control individuals, based on our multiparametric analysis, we built a score from variables derived from [Formula: see text] matrix which showed good performances: area under the receiving operating characteristic curve 0.948 (0.920-0.970), p<0.0001; best discriminating value: negative predictive value 94.7%, positive predictive value was 78.4%. These results seem promising to approach RBC alterations in routine laboratory practice. The related potential clinically relevant outcomes remain to be investigated.

Adipocyte traits limiting cellular insulin responsiveness and glucose transport.

Adipocyte dysfunction is a hallmark of systemic insulin resistance. Insulin-responsive glucose transporter 4 (GLUT4) is downregulated in the insulin resistant state, and cellular insulin responsiveness varies depending on fat depot origin and degree of adipose expansion. Here, we have resolved factors limiting cellular insulin responsiveness, by examining adipocyte function and traits related to glucose transport at the cellular level. Subcutaneous (inguinal) and visceral (epididymal) adipocytes were isolated from C57BL/6J mice fed either chow or high-fat diet. Cell size was determined using coulter counter method, glucose uptake and cytosolic volume were assessed using glucose tracer assays. Total and GLUT4 protein content expressions were determined by Western blot. We found that basal glucose uptake per cell was preserved independent of diet or fat depot origin. Insulin-stimulated glucose uptake per cell was sustained in visceral adipocytes but decreased with adipose expansion in subcutaneous adipocytes. In parallel, the cytosolic space and total protein increased proportionally to total cellular volumetric expansion in visceral, but not in subcutaneous, adipocytes, whereas GLUT4 content decreased exclusively in expanding subcutaneous adipocytes. Together, these data support the existence of distinct phenotypic adipocyte traits that could limit cellular insulin responsiveness. Potentially, these characteristics account for fat depot-specific differences related to glucose transport capacity.NEW & NOTEWORTHY This work illustrates that adipocyte characteristics related to fat depot origin rather than adipocyte size per se limit cellular insulin responsiveness and glucose uptake in male C57BL/6J mice. These findings contribute to the overall understanding of factors limiting adipocyte function and how adipose progression affects insulin response and glucose transport capacity differently in diverse fat depots. Future studies examining whether the proposed characteristics hold true in adipocytes derived from female mice or human origin are needed.