A Molecular Approach to Immunoscintigraphy: A Study of the T-Antigen Conformation on the Surface of Tumors

1987 ◽  
Vol 26 (01) ◽  
pp. 1-6 ◽  
Author(s):  
S. Selvaraj ◽  
M. R. Suresh ◽  
G. McLean ◽  
D. Willans ◽  
C. Turner ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Tumor Cell ◽  
T Antigen ◽  
Human Carcinomas ◽  
Animal Tumor ◽  
Synthetic Antigens

The role of glycoconjugates in tumor cell differentiation has been well documented. We have examined the expression of the two anomers of the Thomsen-Friedenreich antigen on the surface of human, canine and murine tumor cell membranes both in vitro and in vivo. This has been accomplished through the synthesis of the disaccharide terminal residues in both a and ß configuration. Both entities were used to generate murine monoclonal antibodies which recognized the carbohydrate determinants. The determination of fine specificities of these antibodies was effected by means of cellular uptake, immunohistopathology and immunoscintigraphy. Examination of pathological specimens of human and canine tumor tissue indicated that the expressed antigen was in the β configuration. More than 89% of all human carcinomas tested expressed the antigen in the above anomeric form. The combination of synthetic antigens and monoclonal antibodies raised specifically against them provide us with invaluable tools for the study of tumor marker expression in humans and their respective animal tumor models.

Factors affecting in vitro and in vivo antioxidant effects. Experimental conditions and nature of oxidants determine antioxidant efficacy

2021 ◽  
pp. 1-9
Author(s):  
Etsuo Niki
Keyword(s):  
Biological Molecules ◽  
Experimental Conditions ◽  
Factors Affecting ◽  
Beneficial Effects ◽  
Multiple Oxidants ◽  
Antioxidant Effects

Reactive oxygen and nitrogen species have been implicated in the onset and progression of various diseases and the role of antioxidants in the maintenance of health and prevention of diseases has received much attention. The action and effect of antioxidants have been studied extensively under different reaction conditions in multiple media. The antioxidant effects are determined by many factors. This review aims to discuss several important issues that should be considered for determination of experimental conditions and interpretation of experimental results in order to understand the beneficial effects and limit of antioxidants against detrimental oxidation of biological molecules. Emphasis was laid on cell culture experiments and effects of diversity of multiple oxidants on antioxidant efficacy.

DETECTION OF INACTIVATED α2-MACROGLOBULIN (α2M) IN HUMAN PLASMA USING MONOCLONAL ANTIBODIES

1987 ◽  
Author(s):  
J Abbink ◽  
J Nuijens ◽  
C Huijbregts ◽  
E Hack
Keyword(s):  
Monoclonal Antibodies ◽  
Longitudinal Studies ◽  
Human Plasma ◽  
Dextran Sulfate ◽  
Optimal Conditions ◽  
Α2 Macroglobulin ◽  
In Vitro Activation

Monoclonal antibodies (mAbs) were raised against human a2M. Five mAbs that bound to α2M in ELISA were further analyzed by a radioimmunoassay (RIA) for their reaction with three types of α2M: native α2M, chemically inactivated α2M (iα2M) (methylamine treated), and proteolytically iα2M. One mAb reacted with all forms of α2M, while four mAbs bound both forms of ia2M but not native α2M. One of these latter mAbs (Ml) was used to develop a RIA (the Ml-assay) for the detection of iα2M in plasma: Ml coupled to Sepharose is incubated with the plasma to be tested, and bound iα2M is detected by a subsequent incubation with polyclonal 125I-anti-α2M antibodies. As little as 5 ng of iα2M can be detected with this assay in the presence of an excess of native α2M. This assay was then applied to measure inactivation of α2M in vitro and in vivo. In vitro activation of the contact system in plasma by dextran sulfate results in the inactivation of ca 10% of α2M. When blood from normal donors was collected under optimal conditions, about 0.5% of the total α2M content appeared to be iα2M. Longitudinal studies in patients (a.o. with septicaemie, during cardiopulmunary bypass) revealed that increased levels of iα2M occurred sporadically. The Ml-assay appears to be useful to monitor the role of α2M in human diseases.

Fructose Phosphorylation and Dephosphorylation by the Intestinal Mucosa of the Rat During Fructose Absorption

1957 ◽  
Vol 189 (2) ◽  
pp. 301-306 ◽  
Author(s):  
Nicholas M. Papadopoulos ◽  
Joseph H. Roe
Keyword(s):  
Intestinal Mucosa ◽  
Phosphate Esters ◽  
Reduction Techniques ◽  
Mucosal Cells ◽  
Normal Metabolism ◽  
Fasted Rats

The role of phosphorylation in the absorption of fructose from the intestinal tract of the fasted rat by in vitro and in vivo techniques was studied. The authors' method for the determination of fructose phosphate esters was used and these esters were identified by paper chromatography and copper reduction techniques. Buffered homogenate of intestinal mucosa of a fasted rat, mixed with ATP, MgCl2, KF and fructose, when incubated at 30°, showed the formation of fructose-6-phosphate and fructose-1, 6-diphosphate at a rate that corresponded to the decrease in free fructose. The same homogenate, mixed with fructose-1, 6-diphosphate and incubated at 37°, showed the formation of fructose-6-phosphate and free fructose at a rate that corresponded to the decrease in the concentration of the diphosphate ester. Following intraduodenal injection of fructose solution into anesthetized fasted rats, homogenates of the intestinal mucosa showed the presence of fructose-6-phosphate and fructose-1, 6-diphosphate in average concentrations 14 and 5 times, respectively, those found in control muocsa, also concentrations of free fructose in the blood of the portal vein up to 24.6 mg % were observed. The large increase in fructose phosphate esters in the intestinal mucosa, observed after fructose administration, suggests that phosphorylation of sugars in absorption serves a more extensive function than to initiate glycolysis for the normal metabolism of the mucosal cells. The data obtained suggest that phosphorylation and dephosphorylation are functional steps in the absorption of fructose from the alimentary tract of the rat.

THE ROLE OF A 3-0 SULFO GROUP IN THE GLUCOSAMINE RESIDUE OF A SYNTHETIC OLIGOSACCHARIDE FOR THE DETERMINATION OF ANTITHROMBOTIC ACTIONS

1987 ◽  
Author(s):  
J M Walenga ◽  
J Fareed ◽  
M Petitou ◽  
J C Lormeau ◽  
M Samama ◽  
...  
Keyword(s):  
Sulfo Group ◽  
Factor Xa ◽  
Antithrombotic Effect ◽  
Antithrombotic Activity ◽  
Thrombosis Model ◽  
Factor Xa Inhibition

We have previously reported on the antithromboticaction of a chemically synthesized heparin pentasaccharide which exhibits high affinity to anti thrombinIII and sole anti-factor Xa activity. In order to investigate the relative importance of the 3-0 sulfo group of this pentasaccharide, we evaluated the in vitro and in vivo antithrombotic activity of a synthetic pentasccharide devoid of the sulfo group at the third position of the glucosamine residue. In amidolytic and clot-based assays the 3-0 de- sulfated pentasaccharide (3-0-DP) failed to exhibit any antifactor Xa actions at concentrations <100 ug/ml in humanor rabbit plasmas, whereas pentasaccharide showed strong factor Xa inhibition at 1.0 ug/ml IK-=3.2x10 M)and at 10.0 ug/ml in rabbit plasma (K.=9.0×10™7 M). Using a rabbit stasis thrombosis model in which thrombosis was induce by human serum or an activated pro-thrombin complex concentrate, 3-0-DP failed to produce any antithrombotic action in acute intravenous regimens at dosages up to 200 ug/kg. In these two models, pentasaccharide produced >80% inhibition of induced thrombosis. These studies demonstrate the critical importance of the 3-0 sulfo group in this heparin pentasaccharide for the determination of antithrombotic activity, and that in this type of oligosaccharide, anti-factor Xa activity is responsible for producing the antithrombotic effect.

scholarly journals The role of brevican in glioma: promoting tumor cell motility in vitro and in vivo

BMC Cancer ◽  
2012 ◽  
Vol 12 (1) ◽  
Author(s):  
Renquan Lu ◽  
Chengsheng Wu ◽  
Lin Guo ◽  
Yingchao Liu ◽  
Wei Mo ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Cell Motility ◽  
Tumor Cell Motility

The Role of Albumin in Developing Rodent Dental Enamel: A Possible Explanation for White Spot Hypoplasia

1992 ◽  
Vol 71 (6) ◽  
pp. 1270-1274 ◽  
Author(s):  
C. Robinson ◽  
J. Kirkham ◽  
S.J. Brookes ◽  
R.C. Shore
Keyword(s):  
Monoclonal Antibodies ◽  
Calcium Phosphate ◽  
Gel Electrophoresis ◽  
Crystal Size ◽  
Dental Enamel ◽  
Maturation Stage ◽  
Crystallite Growth

The uptake of serum albumin by maturation-stage rodent enamel and the resulting effects on the growth of enamel crystallites were investigated in vitro. Albumin uptake was demonstrated by means of gel electrophoresis and confirmed by Western blotting with use of monoclonal antibodies. Measurement of crystal size was carried out by direct TEM measurement of enamel crystallite outlines after incubations in metastable solutions of calcium phosphate. The ability of endogenous enamel enzymes to degrade albumin was investigated by substrate-specific zymography. The results showed that albumin could be taken up by maturation-stage enamel and produce inhibition of crystallite growth. There was no detectable proteolytic activity in the enamel against albumin substrate, which suggests that albumin entering enamel by extravasation in vivo may produce incomplete tissue maturation, resulting in a white, opaque appearance on eruption.

p125 focal adhesion kinase promotes malignant astrocytoma cell proliferation in vivo

2000 ◽  
Vol 113 (23) ◽  
pp. 4221-4230 ◽  
Author(s):  
D. Wang ◽  
J.R. Grammer ◽  
C.S. Cobbs ◽  
J.E. Stewart ◽  
Z. Liu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Tumor Cell ◽  
Tumor Cells ◽  
Focal Adhesion ◽  
Fold Increase ◽  
Astrocytic Tumor ◽  
Malignant Astrocytoma

p125 focal adhesion kinase (p125FAK) is a cytoplasmic tyrosine kinase that is activated upon engagement of integrin cell adhesion receptors, and initiates several signaling events that modulate cell function in vitro. To determine the biologic role of p125FAK in malignant astrocytic tumor cells, U-251MG human malignant astrocytoma cells were stably transfected with p125FAK cDNA using the TET-ON system, and stable clones isolated that exhibited an estimated 5- or 20-fold increase in p125FAK expression on administration of 0.1 or 2.0 microg/ml doxycycline, respectively. In vitro studies demonstrated that induction of p125FAK resulted in a 2- to 3-fold increase in cell migration, increased p130CAS phosphorylation, localization of exogenous p125FAK to focal adhesions, and a 2-fold increase in soft agar growth. To determine the role of p125FAK in vivo, clones were injected stereotactically into the brains of scid mice. A 4.5-fold estimated increase in p125FAK expression was induced by administration of doxycycline in the drinking water. Analysis of xenograft brains demonstrated that, upon induction of p125FAK, there was a 1.6- to 2.8-fold increase in tumor cell number, and an increase in mAb PCNA-labeling of tumor cells in the absence of a change in the apoptotic index. Compared to normal brain, the expression of p125FAK was elevated in malignant astrocytic tumor biopsies from patient samples. These data demonstrate for the first time that p125FAK promotes tumor cell proliferation in vivo, and that the underlying mechanism is not associated with a reduction in apoptosis.

scholarly journals Overexpression of Cosmc suppresses cell migration and invasion in different subtypes of breast cancer cells via Tn and T glycans

2020 ◽  
Vol 40 (6) ◽  
Author(s):  
Jun Liu ◽  
Feng Xu ◽  
Jie Li ◽  
Hongchuan Jiang
Keyword(s):  
Breast Cancer ◽  
Cell Lines ◽  
T Antigen ◽  
Primary Breast Tumor ◽  
Migration And Invasion ◽  
Related Factors ◽  
In Vivo Experiments

Abstract Objectives: The high mortality of breast cancer (BC) is associated with the strong metastatic properties of primary breast tumor cells. The present study was conducted in order to clarify the effect of Cosmc on the growth and metastasis of BC cell lines of different molecular types, which may be implicated in the regulation of Tn and T glycans. Methods: BC cell lines with different molecular types were transduced with shRNA targeting Cosmc or, Cosmc overexpression plasmid in order to explore the role of Cosmc in cell proliferation, migration, invasion, and apoptosis. The protein levels of Tn, T, Cosmc, proliferation-related factors (Ki67 and PCNA) and apoptosis-related factors (Bax and Bad) in BC cell lines were determined by Western blot analyses. Finally, the role of Cosmc was substantiated through in vivo experiments. Results: Cosmc was down-regulated in different subtypes of BC cell lines compared with normal control cells. Overexpression of Cosmc suppressed the proliferation, migration, and invasion, yet promoted the apoptosis of BC cells, as reflected by in vitro experiments. Additionally, in vivo tumor xenografts in nude mice showed that ectopic overexpression of Cosmc inhibited the tumor growth of BC cells. Consequently, the levels of proliferation-related factors and Tn antigen were decreased, while those of apoptosis-related factors and T antigen were increased in BC cells. This observation was confirmed in vivo in xenograft tumors. Conclusion: Collectively, up-regulation of Cosmc potentially impedes BC growth and metastasis by modulating the balance between Tn and T glycans.

scholarly journals Changes in erythrocyte deformability during day and possible role of melatonin

2018 ◽  
Vol 52 (1) ◽  
pp. 17-20 ◽  
Author(s):  
Rastislav Vazan ◽  
Katarina Plauterova ◽  
Gabriela Porubska ◽  
Jana Radosinska
Keyword(s):  
Erythrocyte Deformability ◽  
Diurnal Changes ◽  
Sample Collection ◽  
Pass Through ◽  
In Vivo Effects ◽  
Vitro Effect

Abstract Objectives. The deformability of erythrocytes is their ability to change shape in order to pass through the capillaries. Th is is necessary for quality of microcirculation and sufficient delivery of oxygen to the tissues. Th e aim of our study was to investigate the possible spontaneous changes in the erythrocyte deformability during day and evaluation of the possible direct effects of melatonin (hormone involved in regulation of biorhythms) on the erythrocyte deformability. Methods. Samples of capillary blood were taken from 12 healthy volunteers in the morning (8:00) and early in the evening (16:30). Determination of erythrocyte deformability was done based on the measurement of their filtrability. It was measured immediately aft er the sample collection and 2-hour lasting incubation without or with melatonin (2000 μmol/L). Results. Erythrocyte deformability was significantly lower in the morning (filtrability index: 0.68±0.01 morning vs. 0.71±0.01 early evening, p<0.05). Th e incubation of blood samples with melatonin did not have impact on deformability. Conclusions. We suggest the presence of diurnal changes in erythrocyte deformability with worse values in the morning that may contribute to higher risk of ischemic attacks in the morning hours. Direct in vitro effect of melatonin on deformability was not observed, but possible in vivo effects cannot be excluded.

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