Putative protective effect of Cadmium chloride highdiluted solution on LLC-PK1 cell intoxicated by high concentration of this same metal: an isopathic in vitro assay.

Cadmium is an important toxic environmental heavy metal. Several studies have demonstrated that a major site of cadmium toxicity in humans and in other animals is the proximal tubule of the kidney. A well established model for nefrotoxicity is the use of in vitro technique with proximal tubule epithelial cell lines, as LLC-PK1. Herein, we have the intention to study the possible protective effect of highdiluted CdCl2 solutions. In a blinding way, LLC-PK1 cells were pre-treated with highdiluted cadmium chloride in the potencies 10 cH, 15 cH and 20cH. After 4 days, these cells have received CdCl2 in a pre-determined toxic concentration. The cell viability was assessed by MTT assay. We have identified a protective effect of two CdCl2 highdiluted solutions, 10 cH and 20 cH, when cells were intoxicated by sublethal CdCl2 concentration. The results indicate that probably the highdilutions have an expressive action on cells in sublethal intoxication.

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Discovery, optimization, and evaluation of non-bile acid FXR/TGR5 dual agonists

Scientific Reports ◽

10.1038/s41598-021-88493-0 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Sachiho Miyata ◽

Yuji Kawashima ◽

Miku Sakai ◽

Masaya Matsubayashi ◽

Keisuke Motoki ◽

...

Keyword(s):

Bile Acid ◽

High Fat Diet ◽

Farnesoid X Receptor ◽

High Concentration ◽

Vitro Assay ◽

Glucose Levels ◽

Blood Glucose Levels ◽

G Protein Coupled ◽

Dual Agonist

AbstractAlthough several potent bile acid Farnesoid X receptor (FXR) and Takeda G-protein-coupled receptor 5 (TGR5, GPBAR1) dual agonists such as INT-767 have been reported, no non-bile acid FXR/TGR5 dual agonist has been investigated to date. Therefore, we attempted to discover potent non-bile acid FXR/TGR5 dual agonists and identified some non-bile acid FXR/TGR5 dual agonists, such as isonicotinamide derivatives in vitro assay. Compound 20p was evaluated in C57BL/6J mice, that were administered a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) consisting of 60 kcal% fat and 0.1% methionine by weight for one week. Compound 20p dose-dependently induced small heterodimer partner (SHP) mRNA and decreased cytochrome P450 7A1 (CYP7A1) in the liver at 10 and 30 mg/kg, respectively, which were used as FXR agonist markers. Compound 20p significantly increased the plasma levels of GLP-1 as a TGR5 agonist, and a high concentration of GLP-1 lowered blood glucose levels. We confirmed that compound 20p was a non-bile acid FXR/TGR5 dual agonist.

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Protective effect of metallothionein on cadmium toxicity in isolated rat hepatocytes

Biochemical Journal ◽

10.1042/bj2300395 ◽

1985 ◽

Vol 230 (2) ◽

pp. 395-402 ◽

Cited By ~ 57

Author(s):

W S Din ◽

J M Frazier

Keyword(s):

Protective Effect ◽

Cadmium Toxicity ◽

Membrane Damage ◽

Rat Hepatocytes ◽

Protective Effects ◽

Isolated Rat Hepatocytes ◽

Detoxification Mechanism ◽

Isolated Rat

An isolated rat hepatocyte preparation was used to study the cellular toxicity of cadmium and the protective effects of metallothionein on cadmium-induced toxicity. Exposure of primary suspension cultures of isolated rat hepatocytes to Cd2+ (0-35.7 microM) for 15 min resulted in a dose-dependent reduction in the synthesis of cellular proteins during a subsequent 6 h incubation. Such inhibition could not be correlated with cellular lethality or gross membrane damage. Pre-induction of metallothionein in hepatocytes by zinc treatment in vivo of donor rats protected hepatocytes in vitro from cadmium-induced inhibition of protein synthesis. The protective effects in zinc-pre-induced hepatocytes are not due to alterations in the level of total cellular cadmium, but could be accounted for by the redistribution of intracellular cadmium in the presence of high levels of zinc-metallothionein. The data suggest that metallothionein exerts its protective effect by a kinetic detoxification mechanism, i.e. a decrease in reactive intracellular cadmium.

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Clinical, Cellular and Molecular Differences Between Newly Diagnosed and Relapsed Patients with Acute Promyelocytic Leukemia: Insights Into Mechanisms of Resistance

Blood ◽

10.1182/blood.v120.21.1390.1390 ◽

2012 ◽

Vol 120 (21) ◽

pp. 1390-1390

Author(s):

Ezhilarasi Chendamarai ◽

Ansu Abu Alex ◽

Saravanan Ganesan ◽

Vandana Kamath ◽

Sukesh C Nair ◽

...

Keyword(s):

Cell Adhesion ◽

Protective Effect ◽

Acute Promyelocytic Leukemia ◽

Stromal Cell ◽

Promyelocytic Leukemia ◽

Differentially Expressed ◽

Taqman Assay ◽

Newly Diagnosed ◽

Vitro Assay

Abstract Abstract 1390 We undertook a comprehensive prospective study to compare the clinical, cellular and molecular differences between newly diagnosed (ND) and relapsed (Rel) acute promyelocytic leukemia (APL) patients. 98 ND cases and 28 Rel cases (5 second relapses) were enrolled in this study. 85 (86.7%) of the ND cases were treated with a single agent arsenic trioxide (ATO) regimen. Five (17.8%) Rel cases were discharged against medical advice, the rest were treated with a combination of ATO, ATRA and chemotherapy and advised a SCT in molecular remission. Table 1 summarizes the major clinical differences between ND and Rel cases. Rel patients had a significantly lower WBC counts and higher platelet counts at diagnosis. Coagulopathy was less prominent in Rel cases as evident by the significantly lower number of cases with IC bleeds in this group and the significantly lower platelet rich concentrate requirement in induction (Table 1). Immunophenotyping of the malignant promyelocytes and blasts revealed a significantly increased CD34 expression and significantly reduced CD13 and CD38 expression in Rel cases (Figure 1A). Cytogenetic (CTG) analysis revealed that relapsed patients had a trend to greater proportion of cases with additional karyotypic abnormalities (Table 1). Evaluation of the malignant promyelocytes and blasts' capacity to concentrate ATO intracellularly as shown in Figure 1B (level of ATO in 107 cells after 24 hour culture with 0.5μM ATO was measured using Atomic Absorption Spectrometry) was not significantly different. Similarly an in-vitro assay of cytotoxicity of ATO (IC-50 measured using standard MTT cell viability assay) was not significantly different between ND and Rel cases (median IC-50= 5.64 ± 3.8 vs. 4.59±4.5 μM; P= 0.28). We had previously noted a protective effect of stromal cell co-culture on the apoptotic action of ATO on malignant promyelocytes. We compared this effect in newly diagnosed and relapsed cases and noted that this protective effect was seen in both groups and was not significantly different (Figure 1C). A microarray expression profile on 8 randomly selected ND and 8 Rel cases was done. Total RNA extracted from BMMNCs with 90% promyelocytes + blasts and above (enriched when required) was used and microarray was performed with a standard one color Agilent–44K array platform. Differentially regulated genes were clustered using hierarchical clustering based on Pearson coefficient correlation, and 1744 genes were 2 fold and above differentially expressed between at relapse and at diagnosis (Figure 1D). 26 differentially expressed genes were validated by RQ-PCR using ABI Taqman assay system (data not shown). Differential gene list were classified based on functional category and pathways using Biointerpreter Software. The most prominent differentially regulated pathways were cell adhesion, cell survival, cell cycle, immune regulation, stem cell regulation, ubiquitinproteasome degradation system and ion transporters. The heat map of the one such prominent pathway (adhesion) is shown in the figure 1E. In conclusion, the Rel cases have significant clinical, immunophenotypic and molecular differences when compared to the newly diagnosed cases. The potential reason for relapse and poor prognosis in the Rel group appears to be multi-factorial. Exploring the pathways and genes differentially regulated between these two groups may provide insights in identifying potentially novel therapeutic targets. Preliminary in-vitro data from our centre, consistent with these observations, has shown that stromal cell adhesion mediates resistance to ATO and that inhibition of this by drugs such as bortezomib can overcome it. Table 1. Comparison of the baseline demographic data of newly diagnosed and relapsed acute promyelocytic leukemia patients Demographic Parameters Newly Diagnosed N=98 N (%)/Median (Range) Relapsed N=28 N (%)/Median (Range) P-value Age (years) 28 (2-60) 31 (8-54) 0.997 Sex: Male 48 (49) 25 (89.3) 0.000 WBC x 109/Lt 14.5 (0.5-290) 3.5 (0.5-24.90) 0.000 Platelet x 109/Lt 17 (3-85) 31.5 (6-248) 0.006 RT-PCR: bcr1 59 (60.2) 21 (75) bcr2 2 (2) 0 0.311 bcr3 37 (37.8) 7 (25) Lactate dehydrogenase (IU/Lt)) 748 (291-2700) 554 (284-1155) 0.006 Fibrinogen mg% 161 (25-689) 163 (70-1122) 0.975 Additional Cytogenetic finding 34 (36.6) 11 (61.1) 0.068 IC Bleed in induction: Yes 16 (16.3) 0 (0%) 0.022 Platelet rich concentrates in induction (units) 24 (0-85) 16 (0-48) 0.027 Disclosures: No relevant conflicts of interest to declare.

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Protective Effect of Shikimic Acid against Cisplatin-Induced Renal Injury: In Vitro and In Vivo Studies

Plants ◽

10.3390/plants9121681 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1681

Author(s):

Jinkyung Lee ◽

Quynh Nhu Nguyen ◽

Jun Yeon Park ◽

Sullim Lee ◽

Gwi Seo Hwang ◽

...

Keyword(s):

Oxidative Stress ◽

Protective Effect ◽

Cell Viability ◽

Renal Injury ◽

Shikimic Acid ◽

Kidney Injury ◽

In Vivo Studies ◽

Artemisia Absinthium ◽

Vitro Assay

Nephrotoxicity is a serious side effect of cisplatin, which is one of the most frequently used drugs for cancer treatment. This study aimed to assess the renoprotective effect of Artemisia absinthium extract and its bioactive compound (shikimic acid) against cisplatin-induced renal injury. An in vitro assay was performed in kidney tubular epithelial cells (LLC-PK1) with 50, 100, and 200 µg/mL A. absinthium extract and 25 and 50 µM shikimic acid, and cytotoxicity was induced by 25 µM cisplatin. BALB/c mice (6 weeks old) were injected with 16 mg/kg cisplatin once and orally administered 25 and 50 mg/kg shikimic acid daily for 4 days. The results showed that the A. absinthium extract reversed the decrease in renal cell viability induced by cisplatin, whereas it decreased the reactive oxidative stress accumulation and apoptosis in LLC-PK1 cells. Shikimic acid also reversed the effect on cell viability but decreased oxidative stress and apoptosis in renal cells compared with the levels in the cisplatin-treated group. Furthermore, shikimic acid protected against kidney injury in cisplatin-treated mice by reducing serum creatinine levels. The protective effect of shikimic acid against cisplatin-mediated kidney injury was confirmed by the recovery of histological kidney injury in cisplatin-treated mice. To the best of our knowledge, this study is the first report on the nephroprotective effect of A. absinthium extract and its mechanism of action against cisplatin-induced renal injury.

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Characterization of Phytochemical Components of Crocus sativus Leaves: A New Attractive By-Product

Scientia Pharmaceutica ◽

10.3390/scipharm89020028 ◽

2021 ◽

Vol 89 (2) ◽

pp. 28

Author(s):

Olha Mykhailenko ◽

Liudas Ivanauskas ◽

Ivan Bezruk ◽

Lyudmila Sidorenko ◽

Roman Lesyk ◽

...

Keyword(s):

Amino Acids ◽

Bioactive Compounds ◽

Essential Amino Acids ◽

Crocus Sativus ◽

Total Phenolic ◽

High Concentration ◽

Vitro Assay ◽

Phytochemical Components ◽

Linoelaidic Acid

Crocus sativus L. is one of the world’s most famous saffron production crops and its enormous by-products, such as leaves, are an excellent source of bioactive compounds with potential nutritional applications. The total phenolic content of Crocus leaves was 5.44 ± 0.01 mg GAE/g, and the total flavonoid content was 2.63 ± 0.05 mg RE/g, respectively. The main bioactive compounds in the leaves, such as polyphenols, flavonoids by HPLC and carboxylic acids, and amino acids, were also identified by GC-MS. HPLC analyses revealed mangiferin as a dominant constituent (1.26 ± 0.02 mg/g). C. sativus contains seven essential amino acids (ILE, LEU, LYS, MET, PHE, THR, TRP, VAL) in high concentration. Among them, isoleucine (7965 µg/g) was the dominant compound. In addition, the K and Ca concentrations in the leaves were significant (p < 0.05). The chemical composition revealed α-linolenic acid (22,490 µg/g) and linoelaidic acid (9880 µg/g) to be major constituents among all the acids found in the Crocus leaves. The extracts of C. sativus leaves showed the highest inhibitory activity for Gram-positive (B. subtilis and S. aureus) bacteria in the in vitro assay. The current results identify and underline the potential of natural products from C. sativus leaves that can add value to saffron production.

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DECOCTION OF POMEGRANATE (PUNICA GRANATUM L.) PEEL AS AN ANTHELMINTIC AGAINST TAENIA SAGINATA

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2021.v13s4.43831 ◽

2021 ◽

pp. 129-131

Author(s):

NYI MEKAR SAPTARINI ◽

RESMI MUSTARICHIE

Keyword(s):

Anthelmintic Activity ◽

Punica Granatum ◽

Positive Control ◽

In Vitro Assay ◽

Taenia Saginata ◽

High Concentration ◽

Pomegranate Peel ◽

Vitro Assay ◽

Punica Granatum L

Objective: The purpose of this study was to investigate the anthelmintic activity of pomegranate (Punica granatum L.) peel decoction against Taenia saginata. Methods: The in vitro assay was conducted by observing the motility of T. saginata, which is isolated from cattle’s gastrointestinal tract, in various concentrations of pomegranate peel decoction with albendazole as a positive control. Results: The results showed that the anthelmintic activity was dependent on decoction concentration and the duration of contact between decoction and nematode. Decoction at moderate concentration causes paralysis, while high concentration causes death. The 75% and 100% pomegranate peel decoction started to cause death at 240 min and 150 min after contact with T. saginata. Conclusion: It was concluded that pomegranate peel decoction has anthelmintic activity.

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Evaluation of Antagonism Activity and Control of Vibrio alginolyticus in Artemia Culture Using Mixed Probiotic

Pertanika Journal of Tropical Agricultural Science ◽

10.47836/pjtas.44.1.07 ◽

2021 ◽

Vol 44 (1) ◽

Author(s):

Mei Yun Beryl Chean ◽

Puvaneswari Puvanasundram ◽

Jasmin Yaminudin ◽

Murni Karim

Keyword(s):

Vibrio Alginolyticus ◽

In Vitro Assays ◽

High Concentration ◽

Immersion Method ◽

Vitro Assay ◽

Single Strain ◽

Artemia Nauplii ◽

And Control

Supplementation with mixed probiotic in aquaculture has been proven to benefit the hosts as disease resistance tool. In this study, a mixed probiotic which consisted of three isolated strains (Lysinibacillus fusiformis strain SPS11, A2, and Bacillus megaterium strain I24) was formulated for the in vitro assays against Vibrio alginolyticus and in vivo preliminary study towards Artemia nauplii. These strains showed antagonism activities against V. alginolyticus in in vitro assay. An increase in biofilm formation of this mixed probiotic was observed which indicated that the strains could work synergistically with each other to confer benefits to the hosts. Enrichment of Artemia nauplii with the formulated mixed probiotic was done to investigate its role in enhancing resistance against the V. alginolyticus. Artemia nauplii were cultured in two different concentrations of mixed probiotic (106 and 108 CFU mL-1) and challenged via immersion method. The mixed probiotic at both concentrations resulted in significantly higher survival of Artemia compared to the challenged group with no probiont added (106 CFU mL-1, 65.00 ± 0.00 % and 108 CFU mL-1, 77.50 ± 3.53 %). Significant reduction of Vibrio loads was observed in Artemia and its culture water supplemented with mixed probiotic at 108 CFU mL-1 whereas there was no reduction of Vibrio at 106 CFU mL-1. This study suggests that the usage of formulated mixed probiotic at high concentration (108 CFU mL-1) as opposed to single-strain probiotic can confer protection against V. alginolyticus infection towards Artemia.

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Weak phenotypic reversion of ivermectin resistance in a field resistant isolate of Haemonchus contortus by verapamil

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2011000900002 ◽

2011 ◽

Vol 31 (9) ◽

pp. 731-736 ◽

Cited By ~ 4

Author(s):

Fernando A. Borges ◽

Julhiano B. Rossini ◽

Patrícia P. Velludo ◽

Carolina Buzzulini ◽

Gustavo H. Costa ◽

...

Keyword(s):

Haemonchus Contortus ◽

Field Isolate ◽

Resistant Isolate ◽

High Concentration ◽

Vitro Assay ◽

Migration Assay ◽

First Results ◽

Phenotypic Reversion

Recent advances in anthelmintic resistant phenotype reversion by Pgp modulating drugs in ruminant nematodes indicate that this can be a useful tool to helminth control. The aim of the present study was to evaluate the efficacy of ivermectin (IVM) in combination with verapamil (VRP), in oil or water-based vehicle, against an IVM-resistant field isolate of Haemonchus contortus through a larval migration assay and experimental infection trial. In the in vitro assay was observed a phenotypic reversion of H. contortus resistance to ivermectin at a high concentration of VRP, increasing IVM efficacy from 53.1% to 94.3. In the in vivo trial, IVM + VRP demonstrated 36.02% efficacy compared to the 7.75% of IVM alone. The vehicle formulation showed no influence in efficacy. These are the first results demonstrating the effect of VRP as a partial IVM-resistance phenotype reverser in a field isolate of IVM-resistant H. contortus experimentally inoculated in sheep.

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Experimental Cell Line Models for Nephrotoxicity Screening

The Bulletin of the Scientific Centre for Expert Evaluation of Medicinal Products ◽

10.30895/1991-2919-2021-11-160-166 ◽

2021 ◽

Vol 11 (3) ◽

pp. 160-166

Author(s):

I. A. Mazerkina ◽

V. A. Evteev ◽

A. B. Prokofiev ◽

O. V. Muslimova ◽

E. Yu. Demchenkova

Keyword(s):

Epithelial Cell ◽

Proximal Tubule ◽

Cell Lines ◽

Test Methods ◽

Renal Proximal Tubule ◽

Cell Models ◽

Proximal Tubule Epithelial Cell ◽

Epithelial Cell Lines

The aim of the study was to review literature data on cell models for experimental assessment of drug nephrotoxicity in vitro. Because of nephrotoxicity, 2% of new investigational medicinal products are discarded at the stage of preclinical in vivo studies in laboratory animals, and 19%—after phase 3 clinical trials. Prediction of toxicity in cell models could make drug development more cost-effective and help to reduce/avoid animal testing. At present, there are no official international guidelines for assessment of nephrotoxicity in vitro, but there is a lot of research underway. The main toxicity target in kidneys is renal proximal tubule epithelial cells, therefore the main research is focused on the development of renal proximal tubule epithelial cell lines with stable functional characteristics. Another important aspect in nephrotoxicity modeling is the choice of relevant test methods and end points which would reflect potential toxicity mechanisms. The paper reviews existing human renal proximal tubule epithelial cell lines and current test methods for assessing cytotoxicity. Promising areas for future development of cell models for nephrotoxicity assessment— are optimisation and standardisation of in vitro systems that would help to make preclinical predictions of drug nephrotoxicity in vivo.

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