scholarly journals Effect of benzyl γ-d-xyloside on the biosynthesis of chondroitin sulphate proteoglycan in cultured human monocytes

1986 ◽  
Vol 238 (1) ◽  
pp. 209-216 ◽  
Author(s):  
S O Kolset ◽  
J Ehlorsson ◽  
L Kjellén ◽  
U Lindahl
Keyword(s):  
Culture Medium ◽  
Regulatory Mechanism ◽  
Core Protein ◽  
Chondroitin Sulphate ◽  
Morphological Changes ◽  
Control Material ◽  
Chondroitin Sulphate Proteoglycan ◽  
Dose Dependent Fashion ◽  
Dose Dependent

Monocytes isolated from human blood differentiate into macrophage-like cells when maintained in vitro for 3-5 days on plastic or glass culture dishes. In the process the cells display characteristic morphological changes, and in addition, a transition in glycosaminoglycan biosynthesis, from the production of chondroitin 4-sulphate to the formation of a polysaccharide containing 20% 4,6-disulphated disaccharide units [Kolset, Kjellén, Seljelid & Lindahl (1983) Biochem. J. 210, 661-667]. Cells were incubated with inorganic [35S]sulphate on day 1 or day 6 in culture, in the presence or absence of benzyl beta-D-xyloside, and labelled polysaccharide was isolated from the culture medium. In the presence of xyloside, the secretion of proteoglycans (90% galactosaminoglycan) was inhibited in a dose-dependent fashion and replaced by release of single polysaccharide chains, the size of which decreased with increasing dose of xyloside. The single polysaccharide chains produced on day 6 in the presence of 0.5 mM-xyloside showed the same proportion of disulphated disaccharide units as did the corresponding control material. Day-1 polysaccharide contained negligible amounts of this component, irrespective of the presence or absence of xyloside. It is concluded that the regulatory mechanism that induces ‘oversulphation’ during the differentiation process operates independently of any association between the polysaccharide chains and the core protein. Moreover, cells maintained in the presence of 0.5 mM-xyloside throughout a 6-day culture period showed the same morphological change, indicative of differentiation into macrophage-like cells, as did untreated control cells. The xyloside did not significantly affect the cytotoxicity of the monocytes, or of the differentiated macrophage-like cells, toward tumour cells.

Soluble Thrombomodulin Purified from Human Urine Exhibits a Potent Anticoagulant Effect In Vitro and In Vivo

1995 ◽  
Vol 73 (05) ◽  
pp. 805-811 ◽  
Author(s):  
Yasuo Takahashi ◽  
Yoshitaka Hosaka ◽  
Hiromi Niina ◽  
Katsuaki Nagasawa ◽  
Masaaki Naotsuka ◽  
...  
Keyword(s):  
Human Urine ◽  
Specific Activity ◽  
Anticoagulant Activity ◽  
Rabbit Lung ◽  
Soluble Thrombomodulin ◽  
Molecular Weights ◽  
Dose Dependent Fashion ◽  
Dose Dependent

SummaryWe examined the anticoagulant activity of two major molecules of soluble thrombomodulin purified from human urine. The apparent molecular weights of these urinary thrombomodulins (UTMs) were 72,000 and 79,000, respectively. Both UTMs showed more potent cofactor activity for protein C activation [specific activity >5,000 thrombomodulin units (TMU)/mg] than human placental thrombomodulin (2,180 TMU/mg) and rabbit lung thrombomodulin (1,980 TMU/mg). The UTMs prolonged thrombin-induced fibrinogen clotting time (>1 TMU/ml), APTT (>5 TMU/ml), TT (>5 TMU/ml) and PT (>40 TMU/ml) in a dose-dependent fashion. These effects appeared in the concentration range of soluble thrombomodulins present in human plasma and urine. In the rat DIC model induced by thromboplastin, administration of UTMs by infusion (300-3,000 TMU/kg) restored the hematological abnormalities derived from DIC in a dose-dependent fashion. These results demonstrate that UTMs exhibit potent anticoagulant and antithrombotic activities, and could play a physiologically important role in microcirculation.

scholarly journals Chondroitin Sulphate Proteoglycan 4 (NG2/CSPG4) Localization in Low- and High-Grade Gliomas

Cells ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 1538 ◽  
Author(s):  
Marta Mellai ◽  
Laura Annovazzi ◽  
Ilaria Bisogno ◽  
Cristiano Corona ◽  
Paola Crociara ◽  
...  
Keyword(s):  
Cell Transformation ◽  
Chondroitin Sulphate ◽  
Molecular Genetic ◽  
Malignant Gliomas ◽  
Malignant Cell ◽  
Wild Type ◽  
Sulphate Proteoglycan ◽  
Chondroitin Sulphate Proteoglycan ◽  
Proteoglycan 4

Background: Neuron glial antigen 2 or chondroitin sulphate proteoglycan 4 (NG2/CSPG4) is expressed by immature precursors/progenitor cells and is possibly involved in malignant cell transformation. The aim of this study was to investigate its role on the progression and survival of sixty-one adult gliomas and nine glioblastoma (GB)-derived cell lines. Methods: NG2/CSPG4 protein expression was assessed by immunohistochemistry and immunofluorescence. Genetic and epigenetic alterations were detected by molecular genetic techniques. Results: NG2/CSPG4 was frequently expressed in IDH-mutant/1p19q-codel oligodendrogliomas (59.1%) and IDH-wild type GBs (40%) and rarely expressed in IDH-mutant or IDH-wild type astrocytomas (14.3%). Besides tumor cells, NG2/CSPG4 immunoreactivity was found in the cytoplasm and/or cell membranes of reactive astrocytes and vascular pericytes/endothelial cells. In GB-derived neurospheres, it was variably detected according to the number of passages of the in vitro culture. In GB-derived adherent cells, a diffuse positivity was found in most cells. NG2/CSPG4 expression was significantly associated with EGFR gene amplification (p = 0.0005) and poor prognosis (p = 0.016) in astrocytic tumors. Conclusion: The immunoreactivity of NG2/CSPG4 provides information on the timing of the neoplastic transformation and could have prognostic and therapeutic relevance as a promising tumor-associated antigen for antibody-based immunotherapy in patients with malignant gliomas.

Papaya shoot tip associated endophytic bacteria isolated from in vitro cultures and host–endophyte interaction in vitro and in vivo

2007 ◽  
Vol 53 (3) ◽  
pp. 380-390 ◽  
Author(s):  
Pious Thomas ◽  
Sima Kumari ◽  
Ganiga K. Swarna ◽  
T.K.S. Gowda
Keyword(s):  
Culture Medium ◽  
Carica Papaya ◽  
In Vitro Cultures ◽  
Dependent Manner ◽  
16S Rdna Sequence ◽  
16S Rdna Sequence Analysis ◽  
Single Organism ◽  
Dose Dependent

Fourteen distinct bacterial clones were isolated from surface-sterilized shoot tips (~1 cm) of papaya (Carica papaya L. ‘Surya’) planted on Murashige and Skoog (MS)-based papaya culture medium (23/50 nos.) during the 2–4 week period following in vitro culturing. These isolates were ascribed to six Gram-negative genera, namely Pantoea ( P. ananatis ), Enterobacter ( E. cloacae ), Brevundimonas ( B. aurantiaca ), Sphingomonas , Methylobacterium ( M. rhodesianum ), and Agrobacterium ( A. tumefaciens ) or two Gram-positive genera, Microbacterium ( M. esteraromaticum ) and Bacillus ( B. benzoevorans ) based on 16S rDNA sequence analysis. Pantoea ananatis was the most frequently isolated organism (70% of the cultures) followed by B. benzoevorans (13%), while others were isolated from single stocks. Bacteria-harboring in vitro cultures often showed a single organism. Pantoea, Enterobacter, and Agrobacterium spp. grew actively on MS-based normal papaya medium, while Microbacterium, Brevundimonas, Bacillus, Sphingomonas, and Methylobacterium spp. failed to grow in the absence of host tissue. Supplying MS medium with tissue extract enhanced the growth of all the organisms in a dose-dependent manner, indicating reliance of the endophyte on its host. Inoculation of papaya seeds with the endophytes (20 h at OD550 = 0.5) led to delayed germination or slow seedling growth initially. However, the inhibition was overcome by 3 months and the seedlings inoculated with Pantoea, Microbacterium, or Sphingomonas spp. displayed significantly better root and shoot growths.

scholarly journals Effect of Lignocaine Concentration on Human Fibroblasts Growth in Eyes Undergoing Trabeculectomy: An in vitro Study

2018 ◽  
Vol 3 (3) ◽  
pp. 1-10 ◽  
Author(s):  
Madhuravasal Krishnan Janani ◽  
Venkatakrishnan Jaichandran ◽  
Hajib Narahari Rao Madhavan ◽  
Lingam Vijaya ◽  
Ronnie Jacob George ◽  
...  
Keyword(s):  
Culture Medium ◽  
In Vitro Model ◽  
Morphological Changes ◽  
Human Fibroblasts ◽  
Annexin V ◽  
In Vitro Study ◽  
Tenon’S Capsule ◽  
Tenon's Capsule ◽  
Vitro Model

Purpose: To evaluate the effect of lignocaine on growth and apoptosis indication of primary human Tenon’s capsule fibroblast (HTFs) in an in vitro model. Patients and Methods: Tenon’s capsule tissue obtained from patients undergoing trabeculectomy were grown in cell culture medium. The effect of different concentrations of lignocaine (0.5, 1.0, 1.5, and 2%) on the morphology and growth of the fibroblasts was studied using microscopy, cell viability, and proliferation assay, and apoptosis was detected using the FITC Annexin V Apoptosis Kit. Results: Morphological changes similar to those of apoptotic cells, including cytoplasmic vacuolation, shrinkage, and rounding were visualized in the cells treated with concentrations greater than 1.0% (i.e., 1.5, 2.0%). Though proliferation inhibition was found with all four concentrations (0.5–2.0%), the viability of cells decreased from 1.0% lignocaine. Conclusion: 0.5% lignocaine prevents proliferation of fibroblasts without causing apoptosis in vitro.

scholarly journals Experimental conversion of virulent RH Toxoplasma gondii tachyzoites in vitro

2001 ◽  
Vol 7 (1-2) ◽  
pp. 181-188
Author(s):  
N. A. Hammouda ◽  
I. R. Ibrahim ◽  
E. D. Elkerdany ◽  
A. Y. Negm ◽  
S. R. Allam
Keyword(s):  
Toxoplasma Gondii ◽  
Culture Medium ◽  
Dna Analysis ◽  
Morphological Changes ◽  
Control Group ◽  
Image Analyser

We aimed to induce conversion of RH-stain tachyzoites to bradyzoites by changing the pH of the culture medium. Alkalization of the medium to pH 8 induced morphological changes in the cultured tachyzoites. The majority of the organism increased in size and changed from a regular crescent shape to a rounded or ovoid shape. Cyst-like structures were formed. Using a computerized image analyser, significant differences in the size of the whole organisms and in their nuclei were observed compared to the control group. The converted organisms also showed significant differences from the control group by quantitative DNA analysis, and did not infect mice.

Paracrystalline bundles of large tubules, induced in vitro by Mebendazole in Cysticercus cellulosae

Parasitology ◽  
1981 ◽  
Vol 83 (3) ◽  
pp. 513-518 ◽  
Author(s):  
J. P. Laclette ◽  
Marie Therese Merchant ◽  
Kaethe Willms ◽  
L. Cañedo
Keyword(s):  
Electron Microscopy ◽  
Culture Medium ◽  
Morphological Changes ◽  
Bladder Wall ◽  
Secretory Cells ◽  
External Diameter ◽  
Nematode Larvae ◽  
Transmission Electron ◽  
Cysticercus Cellulosae

SUMMARYThe effect of the anthelmintic Mebendazole on Cysticercus cellulosae maintained in culture medium was studied by transmission electron microscopy. In addition to the well-known morphological changes induced by Mebendazole in other cestode and nematode larvae, it also induced the cytoplasmic appearance of paracrystalline bundles in the secretory cells of the bladder wall. These bundles were formed by groups of large parallel tubules arranged in a hexagonal-like pattern. The tubules, which had an external diameter of about 50 nm and a length that might exceed 5 μm, were surrounded by a matrix and a distance between neighbouring tubules of 80–120 nm centre to centre was estimated. The tubules were stable to colchicine and low temperature. The temporary appearance of bundles is described and some alternative explanations on their origin are advanced.

scholarly journals Relation of the CD11/CD18 family of leukocyte antigens to the transient neutropenia caused by chemoattractants

Blood ◽  
1990 ◽  
Vol 76 (6) ◽  
pp. 1240-1245 ◽  
Author(s):  
C Lundberg ◽  
SD Wright
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Dose Dependence ◽  
Blood Stream ◽  
Leukocyte Antigens ◽  
Similar Dose ◽  
Inflammatory Site ◽  
Dose Dependent Fashion ◽  
Plasma Leakage ◽  
Dose Dependent

Abstract Adherence of leukocytes to the endothelium is a prerequisite for infiltration and accumulation of cells at an inflammatory site. Recent studies suggest that the CD11/CD18 family of adhesion-promoting receptors plays a crucial role in the initial adherence of polymorphonuclear leukocytes (PMNs) to endothelium. We have studied the effect of the anti-CD18 monoclonal antibody (MoAb) IB4, on movement of PMN in rabbits. Accumulation of PMNs in the skin induced by a local injection of the chemoattractant, zymosan-activated serum (ZAS), was strongly inhibited, in a dose-dependent fashion, by intravenous injection of IB4. A greater than 95% reduction in PMN accumulation was seen with 1 mg IB4/kg body weight, the highest dose used. PMN-dependent plasma leakage in the ZAS-injected skin sites was also inhibited by pretreatment with MoAb IB4, with a similar dose dependence. Histamine- induced plasma leakage, which is PMN independent, was not affected by this treatment. F(ab)2 fragments of IB4 were as effective as the whole immunoglobulin G molecule in reducing PMN accumulation. The half-life of circulating IB4 in rabbits was found to be 11.5 hours. These results are consistent with in vitro studies that show that binding of PMNs to endothelium requires both expression of CD11/CD18 molecules and activation of the PMNs by agonists, and confirm that sites on CD11/CD18 that recognize endothelial cells are blocked by IB4. Other investigators have shown that injection of chemoattractants into the blood stream causes a rapid neutropenia associated with accumulation of PMNs in the lung. We find that intravenous treatment of animals with IB4 did not block the transient accumulation of PMNs in the lung induced by formyl-methionyl-leucyl-phenylalanine, suggesting that this accumulation occurs by a mechanism that does not require CD11/CD18 molecules.

scholarly journals Effects of Caffeoylquinic Acid Derivatives and C-Flavonoid from Lychnophora ericoides on in vitro Inflammatory Mediator Production

2010 ◽  
Vol 5 (5) ◽  
pp. 1934578X1000500 ◽  
Author(s):  
Michel David dos Santos ◽  
Guanjie Chen ◽  
Maria Camila Almeida ◽  
Denis Melo Soares ◽  
Glória Emília Petto de Souza ◽  
...  
Keyword(s):  
Inflammatory Mediators ◽  
Cultured Cells ◽  
Dependent Manner ◽  
Caffeoylquinic Acid ◽  
Inflammatory Mediator Production ◽  
Dicaffeoylquinic Acid ◽  
Tnf Α ◽  
Dose Dependent Fashion ◽  
Dose Dependent

In this study we aimed at evaluating the effect of the major polar constituents of the medicinal plant Lychnophora ericoides on the production of inflammatory mediators produced by LPS-stimulated U-937 cells. The 6,8-di- C-β-glucosylapigenin (vicenin-2) presented no effect on tumor necrosis factor (TNF)-α production, but inhibited, in a dose-dependent manner, the production of prostaglandin (PG) E2 without altering the expression of cyclooxygenase (COX) -2 protein. 3,5-Dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid, at lower concentrations, had small but significant effects on reducing PGE2 levels; at higher doses these compounds stimulated PGE2 and also TNF-α production by the cells. All the caffeoylquinic acid derivatives, in a dose-dependent fashion, were able to inhibit monocyte chemoattractant protein-3 synthesis/release, with 4,5-DCQ being the most potent at the highest tested concentration. These results add important information on the effects of plant natural polyphenols, namely vicenin-2 and caffeoylquinic acid derivatives, on the production of inflammatory mediators by cultured cells.

cAMP-stimulated rise of [Ca2+]i in rabbit connecting tubules: role of peritubular Ca

1990 ◽  
Vol 258 (3) ◽  
pp. F751-F755 ◽  
Author(s):  
J. E. Bourdeau ◽  
B. K. Eby
Keyword(s):  
Parathyroid Hormone ◽  
Ethylene Glycol ◽  
Cell Activation ◽  
Proximal Tubules ◽  
Tetraacetic Acid ◽  
Luminal Membrane ◽  
Dose Dependent Fashion ◽  
Dose Dependent

Parathyroid hormone (PTH) increases cytosolic free Ca concentration ([ Ca2+]i) by mechanisms that depend on extracellular Ca in both cultured renal proximal tubules and isolated rabbit connecting tubules (CNTs). In CNTs 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP) mimics this action, implicating cAMP as a second messenger, and part of the rise, due to increased luminal membrane Ca entry, is likely related to Ca absorption. In cultured proximal tubules the rise in [Ca2+]i, presumably mediated by increased Ca entry across the basolateral plasmalemma, activates gluconeogenesis and shortens microvilli. In the present study we examined cAMP-mediated Ca entry across the basolateral membranes of CNT cells, an effect potentially related to cell activation. Single CNTs were dissected from rabbit kidneys and loaded with fura-2. [Ca2+]i was measured by dual-wavelength excitation during perfusion of isolated segments in vitro. With 1.8 or 2.0 mM Ca in the lumen and the bath, suffusate 8-BrcAMP increased [Ca2+]i within minutes in a dose-dependent fashion. The increase persisted as long as 8-BrcAMP was present and reversed on its withdrawal. With 0.1 microM Ca in the lumen and the bath, 8-BrcAMP, but not ionomycin, failed to increase [Ca2+]i, implying that extracellular Ca is the major source. In tubules perfused with 2 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid to eliminate luminal Ca, but suffused with 1.8 or 2.0 mM Ca, 8-BrcAMP increased [Ca2+]i (though less so than with Ca in the lumen), implying Ca entry across basolateral cell membranes. This rise in [Ca2+]i was attenuated markedly by the presence of 50 microM LaCl3 in the bath.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals A Cumulative Spore Killing Approach: Synergistic Sporicidal Activity of Dilute Peracetic Acid and Ethanol at Low pH Against Clostridium difficile and Bacillus subtilis Spores

2015 ◽  
Vol 3 (1) ◽  
Author(s):  
Michelle M. Nerandzic ◽  
Thriveen Sankar C ◽  
Peter Setlow ◽  
Curtis J. Donskey
Keyword(s):  
Bacillus Subtilis ◽  
Clostridium Difficile ◽  
Peracetic Acid ◽  
Water Washing ◽  
Healthcare Settings ◽  
Synergistic Enhancement ◽  
Spore Killing ◽  
Dose Dependent Fashion ◽  
Dose Dependent

Abstract Background.  Alcohol-based hand sanitizers are the primary method of hand hygiene in healthcare settings, but they lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We previously demonstrated that acidification of ethanol induced rapid sporicidal activity, resulting in ethanol formulations with pH 1.5–2 that were as effective as soap and water washing in reducing levels of C difficile spores on hands. We hypothesized that the addition of dilute peracetic acid (PAA) to acidified ethanol would enhance sporicidal activity while allowing elevation of the pH to a level likely to be well tolerated on skin (ie, >3). Methods.  We tested the efficacy of acidified ethanol solutions alone or in combination with PAA against C difficile and Bacillus subtilis spores in vitro and against nontoxigenic C difficile spores on hands of volunteers. Results.  Acidification of ethanol induced rapid sporicidal activity against C difficile and to a lesser extent B subtilis. The addition of dilute PAA to acidified ethanol resulted in synergistic enhancement of sporicidal activity in a dose-dependent fashion in vitro. On hands, the addition of 1200–2000 ppm PAA enhanced the effectiveness of acidified ethanol formulations, resulting in formulations with pH >3 that were as effective as soap and water washing. Conclusions.  Acidification and the addition of dilute PAA induced rapid sporicidal activity in ethanol. Our findings suggest that it may be feasible to develop effective sporicidal ethanol formulations that are safe and tolerable on skin.

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