scholarly journals Optimization of Blood Handling and Peripheral Blood Mononuclear Cell Cryopreservation of Low Cell Number Samples

2021 ◽  
Vol 22 (17) ◽  
pp. 9129
Author(s):  
Christopher M. Hope ◽  
Dao Huynh ◽  
Ying Ying Wong ◽  
Helena Oakey ◽  
Griffith Boord Perkins ◽  
...  
Keyword(s):  
Storage Temperature ◽  
Viable Cell ◽  
Cell Number ◽  
Blood Collection ◽  
Viable Cell Number ◽  
Peripheral Blood Mononuclear ◽  
Immune Phenotype ◽  
Ifn Γ ◽  
And Function ◽  
Over Time

Background: Rural/remote blood collection can cause delays in processing, reducing PBMC number, viability, cell composition and function. To mitigate these impacts, blood was stored at 4 °C prior to processing. Viable cell number, viability, immune phenotype, and Interferon-γ (IFN-γ) release were measured. Furthermore, the lowest protective volume of cryopreservation media and cell concentration was investigated. Methods: Blood from 10 individuals was stored for up to 10 days. Flow cytometry and IFN-γ ELISPOT were used to measure immune phenotype and function on thawed PBMC. Additionally, PBMC were cryopreserved in volumes ranging from 500 µL to 25 µL and concentration from 10 × 106 cells/mL to 1.67 × 106 cells/mL. Results: PBMC viability and viable cell number significantly reduced over time compared with samples processed immediately, except when stored for 24 h at RT. Monocytes and NK cells significantly reduced over time regardless of storage temperature. Samples with >24 h of RT storage had an increased proportion in Low-Density Neutrophils and T cells compared with samples stored at 4 °C. IFN-γ release was reduced after 24 h of storage, however not in samples stored at 4 °C for >24 h. The lowest protective volume identified was 150 µL with the lowest density of 6.67 × 106 cells/mL. Conclusion: A sample delay of 24 h at RT does not impact the viability and total viable cell numbers. When long-term delays exist (>4 d) total viable cell number and cell viability losses are reduced in samples stored at 4 °C. Immune phenotype and function are slightly altered after 24 h of storage, further impacts of storage are reduced in samples stored at 4 °C.

scholarly journals Application of a roller conveyor type plasma disinfection device with fungus-contaminated citrus fruits

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Akikazu Sakudo ◽  
Yoshihito Yagyu
Keyword(s):  
Treatment Time ◽  
Citrus Fruit ◽  
Viable Cell ◽  
Cell Number ◽  
Viable Cell Number ◽  
Citrus Fruits ◽  
Decimal Reduction Time ◽  
Gas Plasma ◽  
Plasma Device ◽  
Time Required

AbstractEfficient methods to achieve the safe decontamination of agricultural products are needed. Here, we investigated the decontamination of citrus fruits to test the antifungal potential of a novel non-thermal gas plasma apparatus, termed a roller conveyer plasma instrument. This instrument generates an atmospheric pressure dielectric barrier discharge (APDBP) plasma on a set of rollers. Penicillium venetum was spotted onto the surface of the fruit or pericarps, as well as an aluminium plate to act as a control, before performing the plasma treatment. The results showed that viable cell number of P. venetum decreased with a decimal reduction time (D value or estimated treatment time required to reduce viable cell number by 90%) of 0.967 min on the aluminium plate, 2.90 min and 1.88 min on the pericarps of ‘Kiyomi’ (Citrus unshiu × C. sinensis) and ‘Kawano-natsudaidai’ (C. natsudaidai) respectively, and 2.42 min on the surface of ‘Unshu-mikan’ (C. unshiu). These findings confirmed a fungicidal effect of the plasma not only on an abiotic surface (aluminium plate) but also on a biotic surface (citrus fruit). Further development of the instrument by combining sorting systems with the plasma device promises an efficient means of disinfecting citrus fruits during food processing.

scholarly journals Viability and Functional Activity of Cryopreserved Mononuclear Cells

2000 ◽  
Vol 7 (4) ◽  
pp. 714-716 ◽  
Author(s):  
Adriana Weinberg ◽  
Li Zhang ◽  
Darby Brown ◽  
Alejo Erice ◽  
Bruce Polsky ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Functional Activity ◽  
Lymphocyte Proliferation ◽  
Mononuclear Cells ◽  
Cell Number ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
And Function ◽  
Functional Analyses ◽  
Cd4 Cell

ABSTRACT Factors that influence viability and function of cryopreserved peripheral blood mononuclear cells (PBMC) were identified on 54 samples from 27 AIDS Clinical Trial Units. PBMC viability ranged from 1 to 96% with a median of 70%, was higher in laboratories with experienced staff, and was not significantly associated with CD4 cell number. Function of cryopreserved PBMC, measured by lymphocyte proliferation, was associated with viability. Preparations with viability greater than or equal to 70% had consistent proliferative responses and were suitable for functional analyses.

scholarly journals INVESTIGATION ON QUALITY PROPERTIES OF TRADITIONAL BULK BREAD COVERED WITH PROBIOTICS AND SOYBEAN OIL EDIBLE COATING

2020 ◽  
Vol 49 (2) ◽  
pp. 144-153
Author(s):  
R. Amiri Qandashtant ◽  
E. Ataye Salehi ◽  
A. Mohamadi Sani ◽  
M. Mehraban Sangatash ◽  
O. Safari
Keyword(s):  
Soybean Oil ◽  
Whey Protein ◽  
Corn Starch ◽  
Lactobacillus Rhamnosus ◽  
Methyl Cellulose ◽  
Viable Cell ◽  
Cell Number ◽  
Whey Protein Concentrate ◽  
Viable Cell Number ◽  
Quality Properties

Probiotic food products are available at the supermarket commercially, but probiotic bakery products are much less in evidence. In the present study, methyl cellulose (2%), whey protein concentrate (2%), corn starch (1%), and soybean oil at 2, 4, and 6% were used for coating layer on the bulked bread surface, and then the quality properties were studied. The results showed that Lactobacillus rhamnosus GG, as probiotic component of the coating, immobilized in corn starch, whey protein, and methyl cellulose films had enhanced viability throughout shelf-life. The probiotics remained viable for 4 days, maintaining high viable cell number levels. Adding soybean oil at 6% concentration enhanced texture, sensory properties, and image index during storage.

scholarly journals Classification of Five Uremic Solutes according to Their Effects on Renal Tubular Cells

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Takeo Edamatsu ◽  
Ayako Fujieda ◽  
Atsuko Ezawa ◽  
Yoshiharu Itoh
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Progression ◽  
Viable Cell ◽  
Cell Number ◽  
Viable Cell Number ◽  
Cycle Progression ◽  
Tubular Cells ◽  
Renal Tubular Cells ◽  
Uremic Solutes ◽  
Renal Tubular

Background/Aims. Uremic solutes, which are known to be retained in patients with chronic kidney disease, are considered to have deleterious effects on disease progression. Among these uremic solutes, indoxyl sulfate (IS) has been extensively studied, while other solutes have been studied less to state. We conducted a comparative study to examine the similarities and differences between IS,p-cresyl sulfate (PCS), phenyl sulfate (PhS), hippuric acid (HA), and indoleacetic acid (IAA).Methods. We used LLC-PK1 cells to evaluate the effects of these solutes on viable cell number, cell cycle progression, and cell death.Results. All the solutes reduced viable cell number after 48-hour incubation. N-Acetyl-L-cysteine inhibited this effect induced by all solutes except HA. At the concentration that reduced the cell number to almost 50% of vehicle control, IAA induced apoptosis but not cell cycle delay, whereas other solutes induced delay in cell cycle progression with marginal impact on apoptosis. Phosphorylation of p53 and Chk1 and expression of ATF4 and CHOP genes were detected in IS-, PCS-, and PhS-treated cells, but not in IAA-treated cells.Conclusions. Taken together, the adverse effects of PCS and PhS on renal tubular cells are similar to those of IS, while those of HA and IAA differ.

scholarly journals Cantharidin decreased viable cell number in human osteosarcoma U-2 OS cells through G2/M phase arrest and induction of cell apoptosis

2019 ◽  
Vol 83 (10) ◽  
pp. 1912-1923 ◽  
Author(s):  
Chia-Ching Chen ◽  
Fu-Shin Chueh ◽  
Shu-Fen Peng ◽  
Wen-Wen Huang ◽  
Chang-Hai Tsai ◽  
...  
Keyword(s):  
Cell Apoptosis ◽  
Viable Cell ◽  
Cell Number ◽  
Viable Cell Number ◽  
Human Osteosarcoma ◽  
Phase Arrest ◽  
M Phase

Comparison of MTT and ATP-based assays for the measurement of viable cell number

1995 ◽  
Vol 10 (1) ◽  
pp. 29-34 ◽  
Author(s):  
Russell D. Petty ◽  
Lesley A. Sutherland ◽  
Elizabeth M. Hunter ◽  
Ian A. Cree
Keyword(s):  
Viable Cell ◽  
Cell Number ◽  
Viable Cell Number

scholarly journals Nitric oxide suppression of cellular proliferation depends on cationic amino acid transporter activity in cytokine-stimulated pulmonary endothelial cells

2011 ◽  
Vol 300 (4) ◽  
pp. L596-L604 ◽  
Author(s):  
Louis G. Chicione ◽  
Michael R. Stenger ◽  
Hongmei Cui ◽  
Andrea Calvert ◽  
Rebecca J. Evans ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cellular Proliferation ◽  
Viable Cell ◽  
Cell Number ◽  
Viable Cell Number ◽  
Cationic Amino Acid Transporter ◽  
Cationic Amino Acid ◽  
Pulmonary Endothelial Cells ◽  
Viable Cells ◽  
Arginase Ii

Inducible nitric oxide (NO) synthase (iNOS) is a stress response protein upregulated in inflammatory conditions, and NO may suppress cellular proliferation. We hypothesized that preventing l-arginine (l-arg) uptake in endothelial cells would prevent lipopolysaccharide/tumor necrosis factor-α (LPS/TNF)-induced, NO-mediated suppression of cellular proliferation. Bovine pulmonary arterial endothelial cells (bPAEC) were treated with LPS/TNF or vehicle (control), and either 10 mM l-leucine [l-leu; a competitive inhibitor of l-arg uptake by the cationic amino acid transporter (CAT)] or its vehicle. In parallel experiments, iNOS or arginase II were overexpressed in bPAEC using an adenoviral vector (AdiNOS or AdArgII, respectively). LPS/TNF treatment increased the expression of iNOS, arginase II, CAT-1, and CAT-2 mRNA in bPAEC, resulting in greater NO and urea production than in control bPAEC, which was prevented by l-leu. LPS/TNF treatment resulted in fewer viable cells than in controls, and LPS/TNF-stimulated bPAEC treated with l-leu had more viable cells than LPS/TNF treatment alone. LPS/TNF treatment resulted in cleaved caspase-3 and cleaved poly(ADP-ribose) polymerase expression, which was attenuated by l-leu. AdiNOS reduced viable cell number, and treatment of AdiNOS transfected bPAEC with l-leu preserved cell number. AdArgII increased viable cell number, and treatment of AdArgII transfected bPAEC with l-leu prevented the increase in cell number. These data demonstrate that iNOS expression in pulmonary endothelial cells leads to decreased cellular proliferation, which can be attenuated by preventing cellular l-arg uptake. We speculate that CAT activity may represent a novel therapeutic target in inflammatory lung diseases characterized by NO overproduction.

scholarly journals Bone morphogenetic protein (BMP) ligands and receptors in bovine ovarian follicle cells: actions of BMP-4, -6 and -7 on granulosa cells and differential modulation of Smad-1 phosphorylation by follistatin

Reproduction ◽  
2004 ◽  
Vol 127 (2) ◽  
pp. 239-254 ◽  
Author(s):  
Claire Glister ◽  
C Fred Kemp ◽  
Philip G Knight
Keyword(s):  
Granulosa Cells ◽  
Ovarian Follicle ◽  
Viable Cell ◽  
Receptor Activation ◽  
Cell Number ◽  
Activin A ◽  
Follicle Cells ◽  
Viable Cell Number ◽  
Type I ◽  
Antral Follicles

Given the paucity of information on the potential roles of bone morphogenetic proteins (BMPs) in the ruminant ovary we conducted immunolocalization and functional studies on cells isolated from bovine antral follicles. Immunocytochemistry revealed expression of BMP-4 and -7 in isolated theca cells whereas granulosa cells and oocytes selectively expressed BMP-6. All three cell types expressed a range of BMP-responsive type-I (BMPRIB, ActRI) and type-II (BMPRII, ActRII, ActRIIB) receptors supporting autocrine/paracrine roles within the follicle. This was reinforced by functional experiments on granulosa cells which showed that BMP-4, -6 and -7 promoted cellular accumulation of phosphorylated Smad-1 but not Smad-2 and enhanced ‘basal’ and IGF-stimulated secretion of oestradiol (E2), inhibin-A, activin-A and follistatin (FS). Concomitantly, each BMP suppressed ‘basal’ and IGF-stimulated progesterone secretion, consistent with an action to prevent or delay atresia and/or luteinization. BMPs also increased viable cell number under ‘basal’ (BMP-4 and -7) and IGF-stimulated (BMP-4, -6 and -7) conditions. Since FS, a product of bovine granulosa cells, has been shown to bind several BMPs, we used the Biacore technique to compare its binding affinities for activin-A (prototype FS ligand) and BMP-4, -6 and -7. Compared with activin-A (Kd 0.28 ± 0.02 nM; 100%), the relative affinities of FS for BMP-4, -6 and -7 were 10, 5 and 1% respectively. Moreover, studies on granulosa cells showed that preincubation of ligand with excess FS abolished activin-A-induced phosphorylation of Smad-2 and BMP-4-induced phosphorylation of Smad-1. However, FS only partially reversed BMP-6-induced Smad-1 phosphorylation and had no inhibitory effect on BMP-7-induced Smad-1 phosphorylation. These findings support functional roles for BMP-4, -6 and -7 as paracrine/autocrine modulators of granulosa cell steroidogenesis, peptide secretion and proliferation in bovine antral follicles. The finding that FS can differentially modulate BMP-induced receptor activation and that this correlates with the relative binding affinity of FS for each BMP type implicates FS as a potential modulator of BMP action in the ovary.

scholarly journals Influence of IGF-I on adhesion, proliferation, and galectin-1 production in JAr and Jeg-3 choriocarcinoma cell lines

2005 ◽  
Vol 13 (1) ◽  
pp. 7-10 ◽  
Author(s):  
Zanka Bojic-Trbojevic ◽  
Miroslava Jankovic ◽  
Ljiljana Vicovac
Keyword(s):  
Cell Lines ◽  
Solid Phase ◽  
Viable Cell ◽  
Cell Number ◽  
Model Systems ◽  
Viable Cell Number ◽  
Adhesion Assay ◽  
Cell Functions ◽  
Igf I ◽  
Jar Cells

BACKGROUND: JAr and Jeg-3 choriocarcinoma cell lines are model systems for the transformed trophoblast and allow studies of phenotype and regulatory factors for particular cell functions. Both cell lines express the receptor for insulin-like growth factor-I (IGF-I). Effects of IGF-I on adhesion, proliferation and galectin-1 production in JAr and Jeg-3 cells were studied. METHODS: The effects of IGF-I on proliferation and galectin-1 production were examined by thiazolyl blue assay and cell based solid phase assay using polyclonal anti-galectin-1 antibodies. The cell adhesion assay was performed on Matrigel coated wells. Galectin-1 production and localization was examined by immunocytochemistry. RESULTS: IGF-I decreased adhesion of JAr cells to 70% of the control value (p<0.05). Cell treatment with 10 ?g/L of IGF-I significantly increased viable cell number: by 13.5% in JAr and 6% in Jeg-3. Gal-1 was immunolocalized intracellularly and associated with the cell membrane in both cell lines. Production of galectin-1 was significantly increased after treatment with IGF-I compared to control: by 7% in JAr cells and by 16% in Jeg-3 cells (p<0.05). CONCLUSION: The data showed that IGF-I affected adhesion and proliferation of choriocarcinoma cells, depending on the cell line. Both choriocarcinoma cell lines studied here produced galectin-1. The amount of galectin-1 was moderately stimulated by IGF-I.

scholarly journals Reduction of susceptibility to azoles and 5-fluorocytosine and growth acceleration in Candida albicans by glucose in urine

2021 ◽  
Author(s):  
Yoshiki Hiyama ◽  
Toyotaka Sato ◽  
Satoshi Takahashi ◽  
Soh Yamamoto ◽  
Noriko Ogasawara ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Candida Albicans ◽  
Antifungal Susceptibility ◽  
Viable Cell ◽  
Cell Number ◽  
Clinical Isolates ◽  
Viable Cell Number ◽  
Therapeutic Effects ◽  
High Concentration ◽  
Tract Infections

Candida albicans species are causal pathogens for urinary tract infections, vulvovaginitis, and balanitis. Diabetes mellitus is a risk factor for Candida infection. To investigate the potential effects of glucosuria on Candida spp. (C. albicans,C. krusei, and C. glabrata), we investigated the influence of their growth and antifungal susceptibilities by glucose in urine. C. albicans spp. exhibited greater growth in urine with glucose (300 and 3,000 mg/dL) than in plain urine taken from healthy volunteers. After 24 h incubation, the viable cell number was more than 10-fold higher in the urine added 3,000 mg/dL glucose than in plain urine. In antifungal susceptibility, more than 80% of C. albicans clinical isolates increased minimum inhibitory concentrations of azoles (fluconazole, itraconazole, voriconazole, and miconazole) and 5-fluorocytosine with the addition of glucose exceeding their breakpoints. This phenomenon was not observed in clinical isolates of C. krusei and C. glabrata. We observed the growth in the urine to which 3,000 mg/dL glucose was added even in the presence of a 128-fold higher minimum inhibitory concentration of fluconazole. In most of the C. albicans clinical isolates, the mRNA expression of the azole resistance genes ERG11, CDR1, CDR2 and MDR1 increased in glucose-added urine compared with plain urine. In conclusion, the growth ofC. albicans is accelerated and azoles and 5-fluorocytosine become ineffective as a result of a high concentration of glucose in urine. These observations provide valuable information about the clinical course and therapeutic effects of azoles against C. albicans infections in patients with diabetes mellitus and hyperglucosuria.

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