In Vitro Effect of Chlorquine and Picroliv on Plasmodium Berghei Induced Alterations in the Activity of Adenosine Triphosphatase, Aryl Hyrocarbon Hydroxylase Enzymes and Malondialdehyde in spleen Explant Culture

2020 ◽  
Vol 20 ◽  
Author(s):  
Anchal Trivedi ◽  
Aparna Misra ◽  
Esha Sarkar ◽  
Anil K. Balapure
Keyword(s):  
Drug Resistance ◽  
Plasmodium Berghei ◽  
Adenosine Triphosphatase ◽  
Explant Culture ◽  
Need To Evaluate ◽  
Mda Content ◽  
High Level ◽  
Vitro Effect ◽  
Positive Effect

Background: In recent years, great progress has been made in reducing the high level of malaria suffering worldwide. There is a great need to evaluate drug resistance reversers and consider new medicines against malaria. There are many approaches to the development of antimalarial drugs. Specific concerns must be taken in to account in these approaches, in particular there requirement for very in expensive and simple use of new therapies and the need to limit drug discovery expenses. Important ongoing efforts are the optimisation of treatment with available medications, including the use of combination therapy. The production of analogs of known agents and the identification of natural products, the use of compounds originally developed against other diseases, the assessment of overcoming drug resistance and the consideration of new therapeutic targets. Liver and spleen are the important organs which are directly associated with malarial complications. Aim: An analysis the Activity of Adenosine Triphosphatase, Aryl Hyrocarbon Hydroxylase Enzymes and Malondialdehyde in spleen Explant Culture. Objective: To determine in-Vitro Effect of Chlorquine and Picroliv on Plasmodium Berghei Induced Alterations in the Activity of Adenosine Triphosphatase, Aryl Hyrocarbon Hydroxylase Enzymes and Malondialdehyde in spleen Explant Culture. Material and method: 1-Histological preparation of spleen explants for paraplast embedding 2-Biochemicalstudies (Enzymes (Atpase, ALP&GST) and the level of protein, Malondialdehyde (MDA). Result: Splenomegalyis one of the three main diagnostic parameters of malaria infection besides fever and anaemia. Many enzymes present in the liver and spleen may also be altered or liberated under different pathological conditions. Enzymes (ATPase, ALP&GST) and the level of protein, Malondialdehyde (MDA) content was found to increase in the liver and spleen explants during malarial infection. In the liver and spleen derived from parasitized CQ treated animals, the activity of all the above enzymes (ATPase, ALP&GST) and the level of protein & MDA of liver/spleen reversed towards the normal for all the 4or3 days of incubations. Picroliv efficacy decreased with the increment of parasitaemia and at 60%parasitaemia. Conclusion: Alkalinephosphatase (ALP) was found to increase with increasing parasitaemia. After the addition of Picroliv to the medium, a decrement in the activity was observed up to day 4 of culture.A similar positive effect of Picroliv was observed on the ATPase and ALP activity of spleen explants.DNA and protein contents also increased in the parasitized liver cultured in the presence of picroliv.On the contrary, in the spleen explants DNA, protein and MDA content were found to decrease after Picroliv supplementation to the culture medium.

scholarly journals Differential expression of HLA-DR antigens in subsets of human CFU-GM

Blood ◽  
1985 ◽  
Vol 66 (4) ◽  
pp. 788-795 ◽  
Author(s):  
JD Griffin ◽  
KD Sabbath ◽  
F Herrmann ◽  
P Larcom ◽  
K Nichols ◽  
...  
Keyword(s):  
Precursor Cells ◽  
Absolute Increase ◽  
Monocyte Differentiation ◽  
Hla Dr ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Acetate Esterase ◽  
High Level ◽  
Vitro Effect

Abstract Expression of HLA-DR surface antigens by granulocyte/monocyte colony- forming cells (CFU-GM) may be important in the regulation of proliferation of these cells. Using immunological techniques to enrich for progenitor cells, we investigated the expression of HLA-DR in subsets of CFU-GM. “Early” (day 14) CFU-GM express higher levels of HLA- DR than do “late” (day 7) CFU-GM. Among late CFU-GM, cells destined to form monocyte (alpha-naphthyl acetate esterase-positive) colonies express higher levels of HLA-DR than do CFU-GM destined to form granulocyte (chloroacetate esterase-positive) colonies. Because high- level expression of DR antigen was a marker for monocyte differentiation, we examined several lymphokines for their effects on both DR expression and in vitro commitment to monocyte differentiation by myeloid precursor cells. DR antigen density could be increased by more than twofold over 48 hours upon exposure to gamma-interferon (gamma-IFN), whereas colony-stimulating factors had no effect. This was associated with a dose-dependent inhibition of total CFU-GM number, and a relative, but not absolute, increase in the ratio of monocyte colonies to granulocyte colonies. Similarly, in day 7 suspension cultures of purified myeloid precursor cells, gamma-IFN inhibited cell proliferation and increased the ratio of monocytes to granulocytes. Thus, despite the induction of high levels of HLA-DR antigen on precursor cells (a marker of monocyte commitment), the dominant in vitro effect of gamma-IFN was inhibition of granulocyte differentiation.

Molecular Epidemiology of Metronidazole Resistance in a Population of Trichomonas vaginalis Clinical Isolates

2000 ◽  
Vol 38 (8) ◽  
pp. 3004-3009 ◽  
Author(s):  
Lauren J. Snipes ◽  
Pascale M. Gamard ◽  
Elizabeth M. Narcisi ◽  
C. Ben Beard ◽  
Tovi Lehmann ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Trichomonas Vaginalis ◽  
Random Amplified Polymorphic Dna ◽  
Transmitted Disease ◽  
Clinical Isolates ◽  
Nucleotide Position ◽  
Sexually Transmitted ◽  
Metronidazole Resistance ◽  
High Level

Trichomonas vaginalis, the causative agent for human trichomoniasis, is a problematic sexually transmitted disease mainly in women, where it may be asymptomatic or cause severe vaginitis and cervicitis. Despite its high prevalence, the genetic variability and drug resistance characteristics of this organism are poorly understood. To address these issues, genetic analyses were performed on 109 clinical isolates using three approaches. First, two internal transcribed spacer (ITS) regions flanking the 5.8S subunit of the ribosomal DNA gene were sequenced. The only variation was a point mutation at nucleotide position 66 of the ITS1 region found in 16 isolates (14.7%). Second, the presence of a 5.5-kb double-stranded RNAT. vaginalis virus (TVV) was assessed. TVV was detected in 55 isolates (50%). Finally, a phylogenetic analysis was performed based on random amplified polymorphic DNA data. The resulting phylogeny indicated at least two distinct lineages that correlate with the presence of TVV. A band-sharing index indicating relatedness was created for different groups of isolates. It demonstrated that isolates harboring the virus are significantly more closely related to each other than to the rest of the population, and it indicated a high level of relatedness among isolates with in vitro metronidazole resistance. This finding is consistent with the hypothesis that drug resistance toT. vaginalis resulted from a single or very few mutational events. Permutation tests and nonparametric analyses showed associations between metronidazole resistance and phylogeny, the ITS mutation, and TVV presence. These results suggest the existence of genetic markers with clinical implications for T. vaginalisinfections.

scholarly journals Increasing the Magnesium Concentration in Various Dialysate Solutions Differentially Modulates Oxidative Stress in a Human Monocyte Cell Line

Antioxidants ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 319 ◽  
Author(s):  
Carmen Vida ◽  
Julia Carracedo ◽  
Patricia de Sequera ◽  
Guillermo Bodega ◽  
Rafael Pérez ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Human Monocyte ◽  
Magnesium Concentration ◽  
Ros Production ◽  
Mda Content ◽  
Monocyte Cell ◽  
Vitro Effect ◽  
Mg Content ◽  
Dialysis Fluids

Oxidative stress is exacerbated in hemodialysis patients by several factors, including the uremic environment and the use of dialysis fluids (DFs). Since magnesium (Mg) plays a key role in modulating immune function and in reducing oxidative stress, we aimed to evaluate whether increasing the Mg concentration in different DFs could protect against oxidative stress in immunocompetent cells in vitro. Effect of ADF (acetate 3 mM), CDF (citrate 1 mM), and ACDF (citrate 0.8 mM + acetate 0.3 mM) dialysates with Mg at standard (0.5 mM) or higher (1, 1.25, and 2 mM) concentrations were assessed in THP-1 monocyte cultures. Reactive oxygen species (ROS) and malondialdehyde (MDA) levels were quantified under basal and uremic conditions (indoxyl sulfate (IS) treatment). Under uremic conditions, the three DFs with 0.5 mM Mg promoted higher ROS production and lipid damage than the control solution. However, CDF and ACDF induced lower levels of ROS and MDA, compared to that induced by ADF. High Mg concentration (1.25 and/or 2 mM) in CDF and ACDF protected against oxidative stress, indicated by reduced ROS and MDA levels compared to respective DFs with standard concentration of Mg. Increasing Mg concentrations in ADF promoted high ROS production and MDA content. Thus, an increase in Mg content in DFs has differential effects on the oxidative stress in IS-treated THP-1 cells depending on the dialysate used.

scholarly journals Metal Nanoparticles Released from Dental Implant Surfaces: Potential Contribution to Chronic Inflammation and Peri-Implant Bone Loss

Materials ◽  
2019 ◽  
Vol 12 (12) ◽  
pp. 2036 ◽  
Author(s):  
Eriberto Bressan ◽  
Letizia Ferroni ◽  
Chiara Gardin ◽  
Gloria Bellin ◽  
Luca Sbricoli ◽  
...  
Keyword(s):  
Chronic Inflammation ◽  
Dental Implant ◽  
Surrounding Tissue ◽  
Potential Contribution ◽  
Vessel Morphology ◽  
Ti Particles ◽  
High Level ◽  
Vitro Effect

Peri-implantitis is an inflammatory disease affecting tissues surrounding dental implants. Although it represents a common complication of dental implant treatments, the underlying mechanisms have not yet been fully described. The aim of this study is to identify the role of titanium nanoparticles released form the implants on the chronic inflammation and bone lysis in the surrounding tissue. We analyzed the in vitro effect of titanium (Ti) particle exposure on mesenchymal stem cells (MSCs) and fibroblasts (FU), evaluating cell proliferation by MTT test and the generation of reactive oxygen species (ROS). Subsequently, in vivo analysis of peri-implant Ti particle distribution, histological, and molecular analyses were performed. Ti particles led to a time-dependent decrease in cell viability and increase in ROS production in both MSCs and FU. Tissue analyses revealed presence of oxidative stress, high extracellular and intracellular Ti levels and imbalanced bone turnover. High expression of ZFP467 and the presence of adipose-like tissue suggested dysregulation of the MSC population; alterations in vessel morphology were identified. The results suggest that Ti particles may induce the production of high ROS levels, recruiting abnormal quantity of neutrophils able to produce high level of metalloproteinase. This induces the degradation of collagen fibers. These events may influence MSC commitment, with an imbalance of bone regeneration.

scholarly journals Oviductal microvesicles and their effect on in vitro maturation of canine oocytes

Reproduction ◽  
2017 ◽  
Vol 154 (2) ◽  
pp. 167-180 ◽  
Author(s):  
Anna Lange-Consiglio ◽  
Claudia Perrini ◽  
Giulia Albini ◽  
Silvia Modina ◽  
Valentina Lodde ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Multicellular Spheroids ◽  
Maturation Rate ◽  
Oocyte Cytoplasm ◽  
Biphasic Systems ◽  
High Level ◽  
Oviductal Fluid ◽  
Positive Effect

The effect of conditioned medium (CM) or microvesicles (MVs), secreted by multicellular spheroids of oviductal cells, and the involvement of some microRNAs (miRNAs) were investigated in canine oocyte maturation. To generate CM, spheroids were cultured for 3 days. MVs were obtained by ultracentrifugation of CM at 100,000 gand measured for size and concentration by NanoSight instrument. Cumulus-oocyte complexes (COCs) were matured at 38.5°C with 5% CO2and 5% of O2in synthetic oviductal fluid (SOF) in biphasic systems: for 24 h, with 5.0 μg/mL of LH and for other 48 h with 10% oestrous bitch serum. SOF was used as control (CTR) or supplemented with 10% CM or 25–50–75–100–150 × 106 MVs/mL labeled with PKH-26. Results show that multicellular aggregates secreted shedding vesicles. By fluorescence microscopy, the incorporation of labeled MVs was visible only at 72 h in oocyte cytoplasm. These MVs had a positive effect (P < 0.05) on maturation rate (MII) at the concentration of 75 and 100 × 106 MVs/mL compared to CM and CTR (20.34% and 21.82% vs 9.09% and 8.66% respectively). The concentration of 150 × 106 MVs/mL provided only 9.26% of MII. The expression of three specific miRNAs (miR-30b, miR-375 and miR-503) was studied. The lower rate of MII with the higher concentration of MVs is possibly due to the high level of miR-375. In conclusion, the oviductal MVs could be involved in cellular trafficking during oocyte maturation and their possible usein vitrocould facilitate the exploitment of canine reproductive biotechnologies.

Expression of p14ARF in De Novo AML with Normal Karyotype. Implication on Drug Resistance and Survival.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4261-4261
Author(s):  
Esbjorn O. Paul ◽  
Stefan Deneberg ◽  
Soren Lehmann ◽  
Sofia Bengtzén ◽  
Hareth Nahi
Keyword(s):  
Drug Resistance ◽  
De Novo ◽  
P53 Protein ◽  
Normal Karyotype ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Low Levels ◽  
De Novo Aml ◽  
Vitro Effect

Abstract The p53 protein exerts a significant role in growth control of cells and impaired function of p53 by mutation or otherwise is regarded to be important for development of many human cancers. The p53 protein is strongly regulated by the E3-ligase HDM-2 that specifically binds to p53 and causes proteosomal degradation. P14ARF encoded from the INK 4a/ARF gene locus of chromosome 9p, activates the p53 pathway by binding to and inhibiting HDM-2. The objective was to study if the level of mRNA for p14ARF in leukemic cells from patients with AML was related to in vitro drug resistance and clinical outcome. Leukemic cells (>90% pure) isolated from 46 adult patients with normal karyotype de novo AML were incubated with antileukemic drugs (daunorubicin, mitoxantrone, etoposide and Ara-C) cultured for 4 days. The effect was determined by bioluminiscence measuring of intracellular ATP compared to an untreated control. We also tested the activity against PRIMA (p53-dependent reactivation and induction of massive apoptosis), a novel, small molecule shown to activate down regulated p53. mRNA for p14ARF was determined by real time PCR. Results: Patients whose leukemic cells expressed high level of p14ARF mRNA in the leukemic cells (≥0.2 compared to the housekeeping gene) had a significantly better survival compared to those with low level (<0.02). Median survival not obtained compared to 9 months. The mean activity of all tested conventional antileukemic drugs was higher on leukemic cell samples expressing p14ARF mRNA ≥0.2 compared to those with low levels. In contrast, PRIMA exerted significantly higher in vitro effect on leukemic cell samples with low levels of p14ARF mRNA. We conclude that low levels of p14ARF in leukemic cells from patients with normal karyotype AML is a marker for poor prognosis, which may depend on impaired p53 activity causing resistance against conventional antileukemic drugs. Treatment with p53 targeting drugs as PRIMA may be a future possibility to improve the outcome for these patients.

The in vitro effect of hexachlorophene on some enzymes in the liver fluke, Fasciola hepatica L.

Parasitology ◽  
1967 ◽  
Vol 57 (4) ◽  
pp. 665-671 ◽  
Author(s):  
Walborg Thorsell
Keyword(s):  
Adenosine Triphosphatase ◽  
Liver Fluke ◽  
Fasciola Hepatica ◽  
The Other ◽  
Critical Examination ◽  
Enzyme Systems ◽  
Vitro Effect ◽  
Reduced Activity ◽  
Succinate Oxidase

The effect of hexachlorophene, a fasciolicide, on some enzyme systems in the liver fluke was studied. The enzymes were adenosine triphosphatase, succinate oxidase and cholinesterase. Adenosine triphosphatase was activated, whereas succinate oxidase and cholinesterase were inhibited when homogenized flukes were incubated in some concentrations of hexachlorophene. Of the three enzyme systems studied in flukes killed by hexachlorophene, only the succinate system showed reduced activity, whereas the other two showed almost normal activities. The study indicates that biochemical changes are associated with the effect of hexachlorophene on the liver fluke.Thanks are due to Dr C. Grant for critical examination of the language, to Miss Maarja Kippar for valuable help with the experiments and to Mrs Astrid Holmström for the typing of the manuscript.This work was supported by grants A 1108/B 904 and A 1635/B 1299 from Jordbrukets Forskningsråd.

Sign in / Sign up

Export Citation Format

Share Document