Comparison of high -performance thin layer chromatography/UV-densitometry and UV-derivative spectrophotometry for determination of trimetazidine in pharmaceutical formulation

New methods for assaying trimetazidine dihydrochloride on the basis of thin layer chromatography and spectrophotometry are proposed and compared in the paper. In HPTLC/UV-densitometry, separation is achieved by using a mobile phase composed of ammonia-methanol (30:70, V/V) on silica gel HPTLC plates F254. Quantification using a non-linear calibration curve is accomplished by densitometric detection at 230 nm. Derivative spectrophotometric determination of trimetazidine dihydrochloride is carried out from the fourth derivative of the absorbance at 233 nm in peak-zero mode. Statistical comparison led to the conclusion that there is no significant difference between the two studied methods and, moreover, that they demonstrate satisfactory accuracy and precision for routine applications

Differential Action of Silver Nanoparticles on ABCB1 (MDR1) and ABCC1 (MRP1) Activity in Mammalian Cell Lines

Silver nanoparticles (AgNPs), due to their unique properties have been receiving immense attention in recent years. In addition to their antibacterial and antifungal activities, AgNPs also cause apoptosis, mitochondria disfunction, nucleic acid damage and show potent anticancer properties in both multidrug resistance (MDR) and sensitive tumors. The MDR phenomenon, caused by the presence of ATP-binding cassette (ABC) proteins, is responsible for the failure of chemotherapy. Thus, investigating the influence of widely used AgNPs on ABC transporters is crucial. In the present study, we have examined the cytotoxicity of silver nanoparticles of a nominal size of 20 nm (Ag20) on the cell lines of different tissue origins. In addition, we have checked the ATP-binding cassette transporters’ activity and expression under AgNP exposure. The results indicate that Ag20 shows a toxic effect on tested cells, as well as modulating the expression and transport activity of ABC proteins

The patient with 5q minus syndrome and V617F mutation with the presence of ringed erythroblasts meeting the criteria of RARS-T effectively treated with lenalidomide – A case report

5q minus syndrome is a form of myelodysplastic syndrome characterized by the presence of an isolated deletion of long arm of the chromosome 5. Patients with 5q minus respond well to the treatment with lenalidomide. The presence of the JAK2 V617F mutation is a common feature of refractory anemia with ring sideroblasts and marked thrombocytosis. Much less is known about effectiveness of lenalidomide in these patients. We present the patient with 5q minus syndrome and JAK2 V617F mutation accompanied by the presence of ringed erythroblasts meeting the criteria of RARS-T. We could identify only two such patients reported in the literature; no details were given about effectiveness of lenalidomide in that group. We observed good response to the treatment with lenalidomide with transfusion independence 9 months after starting of the treatment; however, there was no complete eradication of del (5)(q13q31) clone nor the clone with JAK V617F mutation

Silver nanoparticles induced changes in DNA methylation and histone H3 methylation in a mouse model of breast cancer

The importance of epigenetic changes as a measurable endpoint in nanotoxicological studies is getting more and more appreciated. In the present work, we analyzed the epigenetic effects induced by citrate- and PEG-coated 20 nm silver nanoparticles (AgNPs) in a model consisting of 4T1 breast cancer tumors in mice. Animals were administered with AgNPs intragastrically (1 mg/kg b.w. daily—total dose 14 mg/kg b.w.) or intravenously (administration twice with 1 mg/kg b.w.—total dose 2 mg/kg b.w.). We observed a significant decrease in 5-methylcytosine (5-mC) level in tumors from mice treated with citrate-coated AgNPs regardless of the route of administration. For PEG-coated AgNPs, a significant decrease in DNA methylation was observed only after intravenous administration. Moreover, treatment of 4T1 tumor-bearing mice with AgNPs decreased histone H3 methylation in tumor tissue. This effect was the most pronounced for PEG-coated AgNPs administered intravenously. No changes in histone H3 Lys9 acetylation were observed. The decrease in methylation of DNA and histone H3 was accompanied by changes in expression of genes encoding chromatin-modifying enzymes (Setd4, Setdb1, Smyd3, Suv39h1, Suv420h1, Whsc1, Kdm1a, Kdm5b, Esco2, Hat1, Myst3, Hdac5, Dnmt1, Ube2b, and Usp22) and genes related to carcinogenesis (Akt1, Brca1, Brca2, Mlh1, Myb, Ccnd1, and Src). The significance of the observed changes and the mechanisms responsible for their development are unclear, and more research in this area is warranted. Nevertheless, the present work points to the epigenetic effects as an important level of interaction between nanomaterials and biological systems, which should always be taken into consideration during analysis of the biological activity of nanomaterials and development of nanopharmaceuticals

Association of genetic dependences between lung cancer and chronic obstructive pulmonary disease

WSTĘP: Dotychczasowe wyniki badań wskazują na zwiększone ryzyko zachorowania na raka płuca u osób ze zmianami obturacyjnymi oskrzeli, w tym u chorych na przewlekłą obturacyjną chorobę płuc (POChP). Wydaje się, że istnieją wspólneczynniki patogenetyczne obu chorób związanych ze zjawiskiem stresu oksydacyjnego. Przedmiotem oceny stały się geny związane z procesami naprawy oksydacyjnych uszkodzeń DNA, geny związane z nowotworami, z metabolizmem żelaza i geny enzymów proteolitycznych.MATERIAŁ I METODY: Badanie przeprowadzono w dwóch grupach pacjentów liczących łącznie 107 osób: 53 chorych na raka niedrobnokomórkowego płuca i POChP oraz 54 chorych tylko na POChP. W przypadku większości genów oznaczono polimorfizm pojedynczego nukleotydu metodą analizy długości fragmentów restrykcyjnych (RFLP). Analizy statystycznej zmiennych ilościowych dokonano testem U Manna-Whitneya i testem median, analiza zmiennych genetycznych została dokonana testem Chi-kwadrat.WYNIKI: W przypadku polimorfizmu genów związanych z metabolizmem żelaza istotne statystycznie różnice pomiędzy badanymi grupami wykazano jedynie w przypadku genu haptoglobiny Hp1/2. Stwierdzono częstsze występowanie formy 1/1 w grupie chorych na POChP i częstsze występowanie formy 1/2 w grupie chorych na raka płuca i POChP. Analiza polimorfizmu genów enzymów proteolitycznych i genu inhibitora tych enzymów wykazała istotne statystycznie różnice pomiędzy badanymi grupami jedynie w przypadku genu metaloproteinazy MMP3 6A/5A. W przypadku polimorfizmu genu metaloproteinazy MMP12 (A-82G) różnice w występowaniu poszczególnych alleli określono na poziomie tendencji.WNIOSKI: Wyniki te wskazują, że u chorych z koincydencją POChP i raka płuca występowały różnice w stosunku do grupy chorych na POChP dotyczące genów związanych z metabolizmem żelaza oraz genów enzymów proteolitycznych.INTRODUCTION: Recent studies have shown an increased risk of lung cancer in patients with bronchial obstructive changes, including patients with COPD. It seems that there are common factors of pathogenesis of both diseases associated with oxidative stress. In the present paper the genes linked to the repair of oxidative damage of DNA, associated with cancer, of iron metabolism and coding proteolytic enzymes were assessed.MATERIAL AND METHODS: The study was conducted in two groups of patients: 53 patients with non-small cell lung cancer and chronic obstructive pulmonary disease, and 54 patients only with chronic obstructive pulmonary disease. The polymorphisms of the single nucleotide were determined in the case of the majority of genes using the PCR-RFLP method. The statistical analysis of quantitative variables was executed using the Mann-Withney U-test and the test of medians; the analysis of genetic variables was executed using the chi² test.RESULTS: Regarding the polymorphisms of genes involved in iron metabolism, statistically significant differences between the two groups have been demonstrated only in the case of haptoglobin gene HP1/2. A higher incidence of form 1/1 was found in patients with COPD and a higher incidence of form 1/2 in patients with lung cancer and COPD. Analysis of gene polymorphisms of proteolytic enzymes and inhibitors of the enzyme gene showed statistically significant differences between the two groups only for the MMP3 gene 6A/5A. In the case of the MMP12 gene polymorphism (A-82G) a tendency toward differences in the occurrence of specific alleles was identified.CONCLUSIONS: These results indicate that patients with coincidence of COPD and lung cancer have disorders of the genes involved in iron metabolism, and they have different genetic polymorphisms of proteolytic enzymes comparing to COPD patients

Matrix metalloproteinase 3 polymorphisms as a potential marker of enhanced susceptibility to lung cancer in chronic obstructive pulmonary disease subjects

[b]Introduction and objective[/b]. Chronic obstructive pulmonary disease (COPD) is often accompanied by lung cancer. Among the genes that may play a role in the occurrence of COPD and lung cancer are those encoding the proteolytic enzymes, such as matrix metalloproteinases (MMPs) and their tissue inhibitors. The objective of this study was to find MMPs-associated markers useful in the identification of COPD subjects with increased susceptibility to developing lung cancer. [b]Materials and methods[/b]. We compared the frequency of single nucleotide polymorphisms in genes coding for matrix proteinases ([i]MMP1, MMP2, MMP3, MMP9, MMP12[/i]) as well as tissue inhibitor of metalloproteinases ([i]TIMP1[/i]) in two groups of subjects: COPD patients (54 subjects) and COPD patients diagnosed for lung cancer occurrence (53 subjects).The levels of the respective proteins in blood serum were also analyzed. [b]Results[/b]. The frequencies of 2 genotypes, [i]MMP3[/i] rs3025058 and MMP3 rs678815, were significantly different between the studied groups. In both cases, more heterozygotes and less homozygotes (both types) were observed in the COPD group than in the COPD + cancer group. A significantly higher TIMP1 level in blood serum was observed in the COPD + cancer group than in the COPD group. There were no statistically significant differences in[i] MMPs[/i] blood levels between the studied groups. In addition, no genotype-associated differences in [i]TIMP1[/i] or[i] MMPs[/i] blood levels were observed. [b]Conclusions[/b]. Homozygocity for [i]MMP3[/i] rs3025058 and rs678815 polymorphisms is a potential marker of enhanced susceptibility to lung cancer development among COPD subjects

Kidney nanotoxicity studied in human renal proximal tubule epithelial cell line TH1

Progressive expansion of nanomaterials in our everyday life raises concerns about their safety for human health. Although kidneys are the primary organs of xenobiotic elimination, little attention has been paid to the kidneys in terms of nanotoxicological studies up to now. Here we investigate the cytotoxic and genotoxic potential of four solid-core uncoated inorganic nanoparticles (TiO₂NPs, SiO₂NPs, Fe₃O₄NPs and AuNPs) using the human renal proximal tubule epithelial TH1 cells. To mimic the in vivo conditions more realistic, TH1 cells were exposed in vitro to inorganic NPs under static as well as dynamic conditions for 3 h and 24 h. The medium throughput alkaline comet assay (12 minigels per slide) was employed to evaluate the impact of these NPs on genome integrity and their capacity to produce oxidative lesions to DNA. The accumulation and localization of studied inorganic NPs inside the cells was monitored by transmission electron microscopy (TEM) and the efficacy of internalization of particular NPs was determined by atomic absorption spectroscopy (AAS) and inductively coupled plasma mass spectrometry (ICP-MS). From all the tested NPs, only Fe₃O₄NPs induced a slight cytotoxicity in TH1 cells exposed to high concentrations (>700 μg/ml) for 24 h. On the other hand, the inorganic NPs did not increase significantly the level of DNA strand breaks or oxidative DNA damage regardless of the treatment mode (static vs. dynamic conditions). Interestingly, substantial differences were observed in the internalized amount of inorganic NPs in TH1 cells exposed to equivalent (2.2 μg/ml) concentration. FeONPs were most efficiently taken up while the lowest quantity of particles was determined in TiONPs-treated cells. As the particle size and shape of individual inorganic NPs in culture medium was nearly identical, it is reasonable to suppose that the chemical composition may contribute to the differences in the efficacy of NPs uptake