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 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 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

Localization of ovine follistatin and α and βA inhibin mRNA in the sheep ovary during the oestrous cycle

1994 ◽  
Vol 12 (2) ◽  
pp. 181-193 ◽  
Author(s):  
D J Tisdall ◽  
N Hudson ◽  
P Smith ◽  
K P McNatty
Keyword(s):  
Gene Expression ◽  
Granulosa Cells ◽  
Ovarian Follicle ◽  
Indirect Evidence ◽  
Oestrous Cycle ◽  
Corpora Lutea ◽  
Α Subunit ◽  
Ovarian Follicle Development ◽  
Antral Follicles

ABSTRACT The sites of follistatin and α and βA inhibin gene expression were examined by in situ hybridization in sheep ovaries during the early and mid-luteal phases (days 3 and 10) of the oestrous cycle and a prostaglandin F2α (PGF2α)-induced follicular phase. Follistatin mRNA was detected in the granulosa cells of preantral, antral and early atretic follicles at all stages of the oestrous cycle, and in the corpora lutea at the early and mid-luteal stages of the cycle. However, only low levels of expression of follistatin were observed in the presumptive preovulatory follicle at 56 h after treatment with PGF2α. Both α and βA inhibin were shown to be expressed in ovaries at all stages of the oestrous cycle. In situ hybridization localized α subunit mRNA to the granulosa cells of most, but not all, healthy antral follicles, and to no other ovarian cell type. In contrast, expression of the βA subunit was confined to a few medium-to-large healthy antral follicles. In antral follicles expressing βA inhibin, mRNAs for α inhibin and follistatin were always detected, but the converse was not true. Unlike follistatin, no α and βA inhibin expression was seen in preantral follicles, developing corpora lutea, or follicles undergoing atresia. These results show that, in the adult sheep ovary, follistatin gene expression is a constitutive event in all growing follicles from the early preantral stage, and also provide indirect evidence of the involvement of follistatin, but not inhibin or activin, in the early stages of ovarian follicle development in sheep.

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 In Vitro Evidence Suggests Activin-A May Promote Tissue Remodeling Associated with Human Luteolysis

Endocrinology ◽  
2007 ◽  
Vol 148 (8) ◽  
pp. 3730-3739 ◽  
Author(s):  
Michelle Myers ◽  
Eva Gay ◽  
Alan S. McNeilly ◽  
Hamish M. Fraser ◽  
W. Colin Duncan
Keyword(s):  
Granulosa Cells ◽  
Primary Cultures ◽  
Gelatin Zymography ◽  
Activin A ◽  
Primary Cell ◽  
Matrix Metalloproteinase 2 ◽  
Corpora Lutea ◽  
Type I ◽  
Rt Pcr ◽  
Maternal Recognition Of Pregnancy

Luteolysis in women is associated with an up-regulation of the expression and activity of matrix metalloproteinase-2 (MMP-2), which is inhibited by human chorionic gonadotropin (hCG) during maternal recognition of pregnancy. Because the primary source of MMP-2 is fibroblasts that do not express LH/hCG receptors, we aimed to investigate the regulation of MMP-2. Women with regular cycles having hysterectomy for nonmalignant conditions and women undergoing oocyte retrieval for assisted conception were used in this current study. Novel primary cultures and cocultures of luteinized granulosa cells and fibroblast-like cells in conjunction with human corpora lutea from different stages of the luteal phase were used to investigate the role of activin-A in the corpus luteum. The effect of hCG, activin-A, and follistatin on MMP-2 activity and expression was assessed by gelatin zymography and quantitative RT-PCR in primary cell cultures. Confirmation of signaling pathways involved in the activation of MMP-2 was assessed by immunofluorescence, RT-PCR, and quantitative RT-PCR. In primary cell culture, steroidogenic cells secrete activin-A and its inhibitors, inhibin-A and follistatin. Follistatin expression is up-regulated by hCG (P < 0.05). The fibroblast-like cells producing MMP-2 have the machinery for activin reception, expressing both type I and type II activin receptors and Smad proteins. Activin-A up-regulated both activity and expression of MMP-2 in fibroblast-like cells (P < 0.05). This activity was inhibited in cocultures of luteinized granulosa cells and fibroblast-like cells in the presence of hCG (P < 0.05) or follistatin (P < 0.01). Activin-A is an excellent candidate for an effector molecule in human luteolysis whose paracrine action is inhibited during maternal recognition of pregnancy.

scholarly journals Activins regulate 17β-hydroxysteroid dehydrogenase type I transcription in murine gonadotrope cells

2009 ◽  
Vol 201 (1) ◽  
pp. 89-104 ◽  
Author(s):  
Beata Bak ◽  
Laura Carpio ◽  
Jinjing L Kipp ◽  
Pankaj Lamba ◽  
Ying Wang ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Intracellular Signaling ◽  
Cell Types ◽  
Hydroxysteroid Dehydrogenase ◽  
Activin A ◽  
Type I ◽  
Promoter Regions ◽  
Gene Encoding ◽  
17Β Estradiol ◽  
Smad Binding Element

Activins are pleiotropic members of the TGFβ superfamily and were initially characterized based on their abilities to stimulate FSH synthesis and secretion by gonadotrope cells of the anterior pituitary gland. Here, we identified the gene encoding the steroidogenic enzyme, 17β-hydroxysteroid dehydrogenase type I (17β-HSD1; Hsd17b1), as an activin-responsive gene in immortalized gonadotrope cells, LβT2. 17β-HSD1 catalyzes the conversion of estrone to the more active 17β-estradiol, and activin A stimulated an increase in this enzymatic activity in these cells. We demonstrated that activins signaled via the type I receptor, activin receptor-like kinase (ALK4), and the intracellular signaling protein, SMAD2, to regulate Hsd17b1 transcription in immediate-early fashion. Critical cis-elements, including a minimal SMAD-binding element, were mapped to within 100 bp of the start of transcription. Activin/ALK4 signaling also regulated Hsd17b1 transcription in both immortalized and primary cultured murine granulosa cells. The promoter regions mediating basal and activin/ALK4-regulated promoter activity were generally conserved across the different cell types. The data show that activin A rapidly regulates Hsd17b1 transcription in gonadotrope and granulosa cells and may thereby regulate local 17β-estradiol synthesis.

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.

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