Antiproliferative and Proapoptotic Activities of Methanolic Extracts from Ligustrum vulgare L. as an Individual Treatment and in Combination with Palladium Complex

The aim of this study is to examine the growth inhibitory effects of methanolic leaf and fruit extracts of L. vulgare on HCT-116 cells over different time periods and their synergistic effect with a Pd(apox) complex. The antiproliferative activity of plant extracts alone or in combination with the Pd(apox) complex was determined using MTT cell viability assay, where the IC50 value was used as a parameter of cytotoxicity. Results show that antiproliferative effects of L. vulgare extracts increase with extension of exposure time, with decreasing IC50 values, except for 72 h where the IC50 values for methanolic leaf extract were lower than for the fruit extract. The Pd(apox) complex alone had a weak antiproliferative effect, but combination with L. vulgare extracts caused stronger effects with lower IC50 values than with L. vulgare extracts alone. The type of cell death was explored by fluorescence microscopy using the acridin orange/ethidium bromide method. Treatments with plant extracts caused typical apoptotic morphological changes in HCT-116 cells and co-treatments with Pd(apox) complex caused higher levels of apoptotic cells than treatment with plant extracts alone. The results indicate that L. vulgare is a considerable source of natural bioactive substances with antiproliferative activity on HCT-116 cells and which have a substantial synergistic effect with the Pd(apox) complex

Cadmium-Induced Oxidative Stress and Apoptotic Changes in the Testis of Freshwater Crab, Sinopotamon henanense

Cadmium (Cd), one of the most toxic environmental and industrial pollutants, is known to exert gonadotoxic and spermiotoxic effects. In the present study, we examined the toxic effect of Cd on the testis of freshwater crab, Sinopotamon henanense. Crabs were exposed to different Cd concentrations (from 0 to 116.00 mg·L−1) for 7 d. Oxidative stress and apoptotic changes in the testes were detected. The activities of SOD, GPx and CAT initially increased and subsequently decreased with increasing Cd concentrations, which was accompanied with the increase in malondialdehyde (MDA) and H2O2 content in a concentration-dependent manner. Typical morphological characteristic and physiological changes of apoptosis were observed using a variety of methods (HE staining, AO/EB double fluorescent staining, Transmission Electron Microscope observation and DNA fragmentation analysis), and the activities of caspase-3 and caspase-9 were increased in a concentration-dependent manner after Cd exposure. These results led to the conclusion that Cd could induced oxidative damage as well as apoptosis in the testis, and the apoptotic processes may be mediated via mitochondria-dependent apoptosis pathway by regulating the activities of caspase-3 and caspase-9

Effects of host defense peptides B2RP, Brevinin-2GU, D-Lys-Temporin, Lys-XT-7 and D-Lys-Ascaphin-8 on peripheral blood mononuclear cells: Preliminary study

© 2017 Croatian Society of Natural Sciences. All rights reserved. Background and purpose: Host defense peptides have considerable therapeutic potential. One of the limitations for their therapeutic use is insufficient selectivity of some peptides, i.e. toxicity for eukaryotic cells. In this study, we have investigated effect of two naturally occurring and three analogs of frog skin-derived peptides on viability/proliferation of resting peripheral blood mononuclear cells and activated lymphocytes. Materials and Methods: Effect of tested peptides was assessed using MTT colorimetric assay. Concanavalin A was used as lymphocyte mitogen. Results: Brevinin-2GU induced cell death only in the highest tested concentration, whereas other peptides were not cytotoxic to resting peripheral blood mononuclear cells. Moreover, high concentrations of B2RP, DLys-Ascaphin-8 and Lys-XT-7 induced cell proliferation and this effect was more prominent in lymphocytes (p<0.05). Tested peptides had opposite effect on activated lymphocytes inhibiting proliferative response to Concanavalin A (Brevinin-2GU, B2RP and D-Lys-Temporin p<0.05). Conclusions: Tested peptides (with exception of Brevinin-2GU) didn’t show cytotoxicity toward peripheral blood mononuclear cells. Moreover, they have potential to modulate immune response by inducing proliferation of resting peripheral blood mononuclear cells and limiting proliferative response to the activation stimulus. Regarding their potent antimicrobial and low hemolytic activity this makes them good candidates for therapeutic use

The cytotoxicity of korbazol against murine cancer cell lines

Background/aim: Korbazol is a natural product that has been shown to exert cytotoxic eff ects on leukemic cells in vitro. The cytotoxicity and biochemical eff ects induced by Korbazol were investigated in the murine cell lines 4T1, B16 and BCL1. Methods: The cytotoxic activity of the tested compound was assessed using a colorimetric MTT assay. The concentration of the superoxide anion radical (O 2.-) and the activity of superoxide dismutase (SOD) in the samples were determined using spectrophotometric methods. The MDA content was determined using a TBA assay. Results: We found that Korbazol induced cell toxicity, an increased the concentration of the lipid peroxidation end product MDA, decreased superoxide dismutase activity and increased superoxide anion formation. Conclusions: Altogether, these results suggest that oxidative stress is involved in Korbazol-induced cytotoxicity in the investigated cell lines

Rsv infection in children aged up to 2 years

Objective. Respiratorysyncytialvirus(RSV) isconsideredto bethemost importantcause ofacuterespiratoryillnessin children. The aim of our paper was to establish the frequency and characteristics of RSV infection in infants with wheezing. Method. We examined a group of 104 infants, diagnosed with wheezing. The concentration of RSV-specific IgG was determined in the serum samples by using ELISA method. Results. We found 24.0% of RSV seropositivechildren in our studygroup, with 12.9% in the first and 47% of RSV IgGserpositive children in the second year of life. Wheezing, as a symptom, was in most cases (61.5%) associated with the diagnosis of bronchitis, whereas in 29.8% of the examined children, it was jointly manifested with two or more other diagnoses. In children diagnosed with bronchitis, the frequency of RSV IgGseropositivity was 20.3%. In those children who had wheezing jointly manifested with two or more diagnoses, the frequency of RSV IgGseropositivity was 32.3%. The greatest percentage of RSV seropositivity (i.e. 40%) was detected in those children who had wheezing and three or more jointly manifested diagnoses. In the group of children who hadhad one episode of wheezing, the frequency of RSV IgGseropositivity was 19.6%, whereas in the group of children with relapsing wheezing (i.e. 4 or more than 4) it was 40.0%. The number of RSV infections wasgreater in female infants in the first year of life and in infants coming from urban environments. Breastfeeding was not found to be a significant factor in prevention of this viral infection. Conclusion. The results of our investigations point to the need of a more effective diagnosis of respiratory infections of viral etiology and further study of local environment- specific risk factors. By knowing them better, we could be able to decrease acute RSV morbidity and chronic consequences of this infection

Suppression of natural killer-cell and dendritic-cell apoptotic tumoricidal activity in patients with head and neck cancer

Background Natural killer (NK) cells and dendritic cells (DCs) mediate tumor cell apoptosis using tumor necrosis factor superfamily ligands (TNFSFLs). This cytotoxicity is an important anticancer immune defense mechanism. Methods We examined TNFSFL expression and apoptotic tumoricidal activity (ATA) of purified NK cells and DCs, and peripheral blood mononuclear leukocytes (PBMLs) of healthy individuals and patients with head and neck cancer (HNC) before and after cancer ablation. Results PBMLs, NK cells and DCs, but not NK-cell/DC-depleted PBMLs, expressed multiple TNFSFLs and mediated ATA. Both TNFSFL expression and ATA were suppressed in tumor-bearing, and restored in tumor-ablated patients with (HNC) Soluble TNF superfamily receptors (solTNFSFRs) were increasingly bound by PBNLs of tumor-bearing HNC patients. Dissociation of solTNFSFR led to more pronounced increases in TNFSFL expression and ATA of PBMLs of patients with HNC than healthy individuals. Conclusion NK-cell and DC TNFSFL expression and ATA are suppressed in patients with HNC. This suppression is tumor-dependent and possibly mediated by solTNFSFRs. © 2012 Wiley Periodicals, Inc

Activity of superoxide dismutase, catalase and glutathione peroxidase in plasma and lysates of chronic lymphocytic leukemia patients

Chronic lymphocytic leukemia is characterized by the progressive accumulation of immature lymphocytes due to inhibited apoptosis. Oxidative stress has a significant role in the pathogenesis of diseases as well as in cancerogenesis itself. The aim of this research was to determine the activity of antioxidative enzymes in blood plasma and in lymphocyte lysates of chronic lymphocytic leukemia patients. Fifty-one patients were divided into two groups according to Rai classification. Group A consisted of patients in stage 0 and stage I and group B consisted of stage II, III and IV patients. Group C consisted of 32 healthy individuals. Enzyme activities of catalase, superoxide dismutase and glutathione peroxidase in lymphocyte lysates of group B patients showed statistically significant decrease. Enzyme activity of catalase in group B patients' blood plasma showed statistically significant increase. Enzyme activities of superoxide dismutase and glutathione peroxidase of group B patients' blood plasma showed no statistically significant changes

Structural, biological and computational study of oxamide derivative

A dicarboxylato-diamide-type compound 2,2'-[(1,2-dioxoethane-1,2-diyl)diimino]dibenzoic acid (H4obbz) (1) was synthesized and characterized. The crystal structure of K2H2obbz·2H2O (2) was determined by X-ray diffracttion analysis. The cytotoxic activities of the compounds were tested against four different cancer cell lines MCF-7, A549, HT-29, HeLa and a human normal cell line MRC-5. The results indicate reasonable dose-dependent cytotoxicity of the ligands that show selectivity against the tested carcinoma and healthy cell lines. Flow cytometric analysis and fluorescence microscopy showed that the most active compound, H4obbz, induced apoptosis and G0/G1 cell cycle arrest, indicating blockage of DNA synthesis as a possible mechanism that triggers apoptosis. Docking and molecular dynamics simulations gave similar responses regarding interactions (binding) between their ligands and chaperon Grp78. The MMGBSA determined ΔG binding energies were in the range from -104 to -140 kJ mol-1,

Molecular diagnosis of bacterial vaginosis – Prevalence of gardnerella vaginalis and atopobium vaginae in pregnant women

© 2018, Serbia Medical Society. All rights reserved. Introduction/Objective Bacterial vaginosis (BV) is defined as disequilibrium of vaginal microbiota due to proliferation of Gram-negative/variable anaerobes and reduction/depletion of vaginal lactobacilli. Difficulties in interpreting microscopically categorized findings in diagnosis of BV need a molecular analysis of bacteria present in vaginal discharge of patients. In this regard, we performed real-time qPCR analysis of vaginal discharge samples with the goal to explore in which extent prevalence and amount of anaerobes, Gardnerella vaginalis and Atopobium vaginae, are related to findings obtained by microscopy. Methods This study enrolled 111 asymptomatic pregnant women between 24 and 28 weeks of pregnancy. Gram-stained vaginal smears were evaluated microscopically. Afterwards, DNA of bacteria was extracted from Gram slides and real-time qPCR was performed with the aim to detect and quantify G. vaginalis and A. vaginae. Results The data of our study showed that 53.2% of patients had normal results, while 20.7% and 26.1% of patients had intermediary (IMD) and BV results, respectively. G. vaginalis and A. vaginae were more frequently found in IMD and BV than in healthy patients; also, the average bacterial number of G. vaginalis and A. vaginae were significantly higher in BV and IMD than in the group with normal findings (p = 0.000). Comparing mutual relation of G. vaginalis and A. vaginae, the prevalence and number of G. vaginalis were in all groups significantly higher than A. vaginae. Conclusion The data of our study have shown that in distinguishing normal from BV findings, quantification of bacteria may be more important than just molecular detection of bacteria