scholarly journals Artifacts in Intracellular Enzyme Distribution Caused by Alkaline Homogenates

1969 ◽  
Vol 44 (3) ◽  
pp. 461-462 ◽  
Author(s):  
P. C. Brandon
Keyword(s):  
Intracellular Enzyme ◽  
Enzyme Distribution

scholarly journals THE ISOLATION OF CELL NUCLEI IN NON-AQUEOUS MEDIA

1952 ◽  
Vol 35 (3) ◽  
pp. 529-557 ◽  
Author(s):  
V. Allfrey ◽  
H. Stern ◽  
A. E. Mirsky ◽  
H. Saetren
Keyword(s):  
Citric Acid ◽  
Serum Proteins ◽  
Aqueous Media ◽  
Organic Media ◽  
Cell Nuclei ◽  
Intracellular Enzyme ◽  
Cytoplasmic Proteins ◽  
Immunological Tests ◽  
Avian Erythrocytes ◽  
Enzyme Distribution

1. A modified Behrens procedure is described for the isolation of nuclei from avian erythrocytes and from the liver, kidney, thymus, pancreas, heart, and intestinal mucosa of the calf or horse. 2. The purity of these nuclei has been established by staining reactions, enzyme studies, and immunological tests for serum proteins. 3. Evidence is presented to show that a transport of cytoplasmic proteins into the nucleus does not occur during the isolation. 4. Nuclei prepared in non-aqueous media contain considerably more protein and a very different enzyme composition from that observed in nuclei prepared by "homogenization" techniques in dilute citric acid. 5. The suitability of nuclei prepared in organic media for the study of intracellular enzyme distribution is discussed.

Design, Synthesis and Biological Evaluation of Novel Urea and Thiourea Bearing thieno[3,2-d]-pyrimidines as PI3 Kinase Inhibitors

2018 ◽  
Vol 18 (6) ◽  
pp. 891-902 ◽  
Author(s):  
Srinu Bodige ◽  
Parameshwar Ravula ◽  
Kali Charan Gulipalli ◽  
Srinivas Endoori ◽  
J.N. Narendra Sharath Chandra ◽  
...  
Keyword(s):  
Anticancer Activity ◽  
Cell Lines ◽  
Inhibitory Activity ◽  
Intracellular Signaling ◽  
Research Work ◽  
Docking Studies ◽  
Biological Evaluation ◽  
Intracellular Enzyme ◽  
Ht 29 ◽  
Mcf 7

Background: Phosphatidylinositol-3-kinase α (PI3Kα) is a ubiquitous intracellular enzyme, mainly involved in intracellular signaling pathways, promotes cellular growth, proliferation, and differentiation. Therefore, inhibition of PI3K can be a hotspot in molecular targeted therapy for the treatment of cancer. Methods: The present research work involves molecular docking studies performed to screen derivatives of urea and thiourea bearing thieno [3,2-d]-pyrimidines against the active site of PI3K enzyme using MOE.2008.10. The designed structures (6a-f) and (7a-j) were synthesized by the facile synthetic methods and evaluated for their anticancer activity against HT-29 and MCF-7 cell lines and inhibitory activity against PI3Kα enzyme. Results: Among the tested compounds, 4-(4-(2-(3-(pyrimidin-2-yl)thioureido)ethyl)piperazin-1-yl)thieno[3,2- d]pyrimidine-6-carboxamide (7f) showed the highest anticancer activity against HT-29 and MCF-7 cell lines with IC50 values of 2.18 µM and 4.25 µM, respectively. Further, the same compound also exhibited potent PI3Kα inhibitory activity with IC50 value of 1.26 µM. Conclusion: Docking studies supported the initial pharmacophoric hypothesis and suggested a mode of interaction at the active binding site of PI3Kα, demonstrating that the target compounds were potential inhibitory agents for cancer therapy.

scholarly journals Whole cell-based catalyst for enzymatic production of the osmolyte 2-O-α-glucosylglycerol

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Katharina N. Schwaiger ◽  
Monika Cserjan-Puschmann ◽  
Gerald Striedner ◽  
Bernd Nidetzky
Keyword(s):  
Specific Activity ◽  
Dry Mass ◽  
High Yield ◽  
Sucrose Phosphorylase ◽  
Bifidobacterium Adolescentis ◽  
Enzymatic Production ◽  
Intracellular Enzyme ◽  
Whole Cell ◽  
Freeze Thaw ◽  
Thaw Cycle

Abstract Background Glucosylglycerol (2-O-α-d-glucosyl-sn-glycerol; GG) is a natural osmolyte from bacteria and plants. It has promising applications as cosmetic and food-and-feed ingredient. Due to its natural scarcity, GG must be prepared through dedicated synthesis, and an industrial bioprocess for GG production has been implemented. This process uses sucrose phosphorylase (SucP)-catalyzed glycosylation of glycerol from sucrose, applying the isolated enzyme in immobilized form. A whole cell-based enzyme formulation might constitute an advanced catalyst for GG production. Here, recombinant production in Escherichia coli BL21(DE3) was compared systematically for the SucPs from Leuconostoc mesenteroides (LmSucP) and Bifidobacterium adolescentis (BaSucP) with the purpose of whole cell catalyst development. Results Expression from pQE30 and pET21 plasmids in E. coli BL21(DE3) gave recombinant protein at 40–50% share of total intracellular protein, with the monomeric LmSucP mostly soluble (≥ 80%) and the homodimeric BaSucP more prominently insoluble (~ 40%). The cell lysate specific activity of LmSucP was 2.8-fold (pET21; 70 ± 24 U/mg; N = 5) and 1.4-fold (pQE30; 54 ± 9 U/mg, N = 5) higher than that of BaSucP. Synthesis reactions revealed LmSucP was more regio-selective for glycerol glycosylation (~ 88%; position O2 compared to O1) than BaSucP (~ 66%), thus identifying LmSucP as the enzyme of choice for GG production. Fed-batch bioreactor cultivations at controlled low specific growth rate (µ = 0.05 h−1; 28 °C) for LmSucP production (pET21) yielded ~ 40 g cell dry mass (CDM)/L with an activity of 2.0 × 104 U/g CDM, corresponding to 39 U/mg protein. The same production from the pQE30 plasmid gave a lower yield of 6.5 × 103 U/g CDM, equivalent to 13 U/mg. A single freeze–thaw cycle exposed ~ 70% of the intracellular enzyme activity for GG production (~ 65 g/L, ~ 90% yield from sucrose), without releasing it from the cells during the reaction. Conclusions Compared to BaSucP, LmSucP is preferred for regio-selective GG production. Expression from pET21 and pQE30 plasmids enables high-yield bioreactor production of the enzyme as a whole cell catalyst. The freeze–thaw treated cells represent a highly active, solid formulation of the LmSucP for practical synthesis.

scholarly journals Biodegradation and metabolic pathway of fenvalerate by Citrobacter freundii CD-9

AMB Express ◽  
2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Jie Tang ◽  
Dan Lei ◽  
Min Wu ◽  
Qiong Hu ◽  
Qing Zhang
Keyword(s):  
Environmental Pollution ◽  
Environmental Remediation ◽  
High Efficiency ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Citrobacter Freundii ◽  
16S Rrna Sequence ◽  
Intracellular Enzyme ◽  
Pyrethroid Insecticides ◽  
Spectrometry Analysis

Abstract Fenvalerate is a pyrethroid insecticide with rapid action, strong targeting, broad spectrum, and high efficiency. However, continued use of fenvalerate has resulted in its widespread presence as a pollutant in surface streams and soils, causing serious environmental pollution. Pesticide residues in the soil are closely related to food safety, yet little is known regarding the kinetics and metabolic behaviors of fenvalerate. In this study, a fenvalerate-degrading microbial strain, CD-9, isolated from factory sludge, was identified as Citrobacter freundii based on morphological, physio-biochemical, and 16S rRNA sequence analysis. Response surface methodology analysis showed that the optimum conditions for fenvalerate degradation by CD-9 were pH 6.3, substrate concentration 77 mg/L, and inoculum amount 6% (v/v). Under these conditions, approximately 88% of fenvalerate present was degraded within 72 h of culture. Based on high-performance liquid chromatography and gas chromatography-mass spectrometry analysis, ten metabolites were confirmed after the degradation of fenvalerate by strain CD-9. Among them, o-phthalaldehyde is a new metabolite for fenvalerate degradation. Based on the identified metabolites, a possible degradation pathway of fenvalerate by C. freundii CD-9 was proposed. Furthermore, the enzyme localization method was used to study CD-9 bacteria and determine that its degrading enzyme is an intracellular enzyme. The degradation rate of fenvalerate by a crude enzyme solution for over 30 min was 73.87%. These results showed that strain CD-9 may be a suitable organism to eliminate environmental pollution by pyrethroid insecticides and provide a future reference for the preparation of microbial degradation agents and environmental remediation.

scholarly journals LDH as a prognostic marker in hypertensive pregnancy

2017 ◽  
Vol 6 (6) ◽  
pp. 2444
Author(s):  
Prathap Talwar ◽  
Triveni Kondareddy ◽  
Pranidha Shree C. A.
Keyword(s):  
Lactic Dehydrogenase ◽  
High Serum ◽  
Pregnancy Induced Hypertension ◽  
Intracellular Enzyme ◽  
Cellular Death ◽  
Severity Of Disease ◽  
Induced Hypertension ◽  
Prospective Comparative Study ◽  
Severity Of The Disease ◽  
Poor Outcomes

Background: Pregnancy induced hypertension (PIH) is a global problem with a 5-15% incidence rate in India and complicating 10-17% of all pregnancies. These are multisystem disorders and lead to a lot of cellular death. LDH is an intracellular enzyme and its level is increased in these women due to cellular death. So, serum LDH levels can be used to assess the extent of cellular death and thereby the severity of disease in this group of women. The objective of the study was to correlate the severity of the disease, maternal and perinatal outcome with Lactic Dehydrogenase (LDH) levels in serum in patients of preeclampsia and eclampsia.Methods: A prospective comparative study was conducted in the department of Obstetrics and Gynecology, JSS Medical Hospital, Mysore.Results: LDH levels were significantly elevated in women with preeclampsia and eclampsia (<0.001). Higher LDH levels had significant correlation with high blood pressure (P <0.10) as well as poor maternal and perinatal outcome.Conclusions: High serum LDH levels correlate well with the severity of the disease and poor outcomes in patients of preeclampsia and eclampsia.

scholarly journals Lactate dehydrogenase levels in preeclampsia and its correlation with maternal and perinatal outcome

2019 ◽  
Vol 8 (4) ◽  
pp. 1505
Author(s):  
Nirmala Bhandari ◽  
Anjali Gupta ◽  
Simmi Kharb ◽  
Meenakshi Chauhan
Keyword(s):  
Lactate Dehydrogenase ◽  
Pregnant Women ◽  
Perinatal Outcome ◽  
Serum Lactate Dehydrogenase ◽  
Hypertensive Disorder ◽  
Perinatal Complications ◽  
Maternal Complications ◽  
Singleton Pregnancy ◽  
Intracellular Enzyme ◽  
Potential Marker

Background: Hypertensive disorder of pregnancy occurs in approximately 6-8% of all pregnancies. The most serious consequences for the mother and the baby are the result of preeclampsia and eclampsia. Lactate Dehydrogenase (LDH) is an intracellular enzyme. Recently LDH has been suggested as potential marker to predict severity of pre-eclampsia. The objective of the present study was to compare the serum lactate dehydrogenase levels in women with preeclampsia and normal pregnant women and to correlate lactate dehydrogenase levels with maternal and perinatal outcome in preeclampsia.Methods: An observational prospective study was conducted on 200 antenatal women attending the labour room emergency. Women with singleton pregnancy and cephalic presentation, from 28 weeks onwards were enrolled in the study. Out of 200, 100 were normal pregnant women and 100 were preeclamptic women. Serum LDH levels were measured in all women and maternal and perinatal outcome was assessed in terms of LDH levels.Results: Higher levels of LDH was observed in pregnant women with preeclampsia (627.38±230.04 IU/l) as compared to normal pregnant women (224.43±116.61 IU/l). The maternal complications were found to be maximum in women with LDH > 800 IU/l.  Abruption was the most common complication. The perinatal mortality and neonatal deaths were found to have significant correlation with high LDH levels.Conclusions: Maternal and perinatal complications were associated with higher LDH levels in preeclampsia patients. Serum LDH levels can be offered to all patients of preeclampsia and can be used to predict the prognosis of preeclampsia.

Effect of enzyme distribution in immobilized membranes

1991 ◽  
Vol 15 ◽  
pp. 249-250
Author(s):  
S. Kunugi ◽  
T. Nakajima ◽  
A. Nomura
Keyword(s):  
Enzyme Distribution

scholarly journals AIE-Driven Fluorescent Polysaccharide Polymersome as Enzyme-responsive FRET Nanoprobe to Study the Real-time Delivery Aspects in Live-Cells

2021 ◽  
Author(s):  
Nilesh Umakant Deshpande ◽  
Mishika Virmani ◽  
Manickam Jayakannan
Keyword(s):  
Energy Transfer ◽  
Confocal Microscopy ◽  
Real Time ◽  
Fluorescence Resonance Energy Transfer ◽  
Imaging Techniques ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Live Cells ◽  
Intracellular Enzyme ◽  
Bio Imaging

We report aggregation induced emission (AIE) driven polysaccharide polymersome as fluorescence resonance energy transfer (FRET) nanoprobes to study their intracellular enzyme-responsive delivery by real-time live-cell confocal microscopy bio-imaging techniques. AIE...

scholarly journals SIMILARITY OF THE KINETICS OF INVERTASE ACTION IN VIVO AND IN VITRO. II

1932 ◽  
Vol 16 (2) ◽  
pp. 233-242 ◽  
Author(s):  
B. G. Wilkes ◽  
Elizabeth T. Palmer
Keyword(s):  
Physiological Activity ◽  
Outer Region ◽  
Living Cells ◽  
Yeast Cells ◽  
Live Cells ◽  
Intracellular Enzyme ◽  
Relationship Of ◽  
Kinetics Of

1. The pH-activity relationship of invertase has been studied in vivo and in vitro under identical external environmental conditions. 2. The effect of changing (H+) upon the sucroclastic activity of living cells of S. cerevisiae and of invertase solutions obtained therefrom has been found, within experimental error, to be identical. 3. The region of living yeast cells in which invertase exerts its physiological activity changes its pH freely and to the same extent as that of the suspending medium. It is suggested that this may indicate that this intracellular enzyme may perform its work somewhere in the outer region of the cell. 4. In using live cells containing maltase, no evidence of increased sucroclastic activity around pH 6.9, due to the action of Weidenhagen's α-glucosidase (maltase), was found.

Sign in / Sign up

Export Citation Format

Share Document