Ligustrum vulgare leaves and fruit extracts induce apoptosis of human leukemia cells

Background and purpose: Cytotoxic effects of Ligustrum vulgare leaves on HeLa cervical tumor cells suggested that Ligustrum vulgare extracts should be investigated as potential anticancer agents. Therefore, we examined a potential antileukemic activity of methanolic extracts of Ligustrum vulgare leaves and fruit extracts on two types of leukemia cells, MOLT-4 and JVM-13, lymphocytes isolated from the blood of 33 chronic lymphocytic leukemia (CLL) patients and on mononuclear leukocytes isolated from the blood of 18 healthy individuals.Material and methods: The cytotoxicity of examined extracts was measured by MTT assay and LDH activity test. The antiapoptotic potential of tested extracts was measured by Annexin V/7AAD flowcytometric assay.Results: The results showed that both extracts exhibited a moderate cytotoxic effect on all three types of leukemia cells. The Ligustrum vulgare leaf extract was the most effective on MOLT-4 cells, the fruit extract on JVM-13 cells and both extracts were equally effective on CLL cells. In addition, none of the tested extracts was toxic to healthy mononuclear cells. Both extracts acted by inducing apoptosis of leukemic cells.Conclusion: Ligustrum vulgare extracts exhibit significant antileukemic potential and should be further investigated

Procjena kromosomske nestabilnosti u limfocitima periferne krvi bolesnika s poremećajima reproduktivnog sustava pomoću mikronukleus-testa

We investigated chromosomal instability in peripheral blood lymphocytes (PBL) of patients with reproductive failure in respect to age, smoking habits, gender, miscarriages, and semen parameters. The study involved 36 individual cases of reproductive failure (18 men and 18 women) attended at the Clinical Centre of Kragujevac, Serbia, and 30 healthy subjects (15 men and 15 women). Micronuclei (MN) frequency was estimated in PBL using the cytokinesis-block micronucleus (CBMN) assay. The baseline MN frequencies were significantly higher (p=0.031; p<0.001) in male [(9.22 ± 4.70) MN per 1000 BN cells] and female patients [(13.50 ± 2.5) MN per 1000 BN cells] than in male and female healthy controls [(6.27 ± 2.66) MN per 1000 BN cells; (6.80 ± 2.98) MN per 1000 BN cells]. The mean baseline MN frequency did not significantly differ between miscarriage groups and between patients with and without normal values of semen parameters. The correlations between poor sperm concentration (<20x106 mL-1), rapid progressive motility (<25 %), normal morphology (<30 %), and MN frequencies were negative, but not statistically significant. We found that only gender significantly influenced the MN rates in analysed patients. There were no significant differences between age groups and between smokers and non-smokers in patients and control samples. We conclude that the increase in baseline MN frequency in PBL of patients with reproductive failure corresponds to the increase in chromosomal damage, which occurs as a result of complex events that cause reproductive disorders.Ispitivali smo kromosomsku nestabilnost u limfocitima periferne krvi (engl. peripheral blood lymphocytes - PBL) bolesnika s poremećajima reproduktivnog sustava u odnosu na parametre dobi, navike pušenja, spola, spontanih pobačaja i kvalitete sjemena. Ispitivanje je uključivalo 36 pojedinačnih ispitanika s poremećajima reproduktivnog sustava (18 muškaraca i 18 žena) u Kliničkom centru u Kragujevcu, Srbiji, te 30 zdravih ispitanika (15 muškaraca i 15 žena). Učestalost pojave mikronukleusa (MN) utvrđena je u PBL-ima primjenom mikronukleus-testa s tehnikom blokirane citokineze (engl. cytokinesis-block micronucleus - CBMN). Učestalosti MN bile su značajno povišene (p=0,031; p<0,001) kod muških [(9,22±4,70) MN na 1000 BN stanica] i ženskih bolesnika [(13,50±2,5) MN na 1000 BN stanica] u odnosu na osnovne vrijednosti utvrđene u muških i ženskih zdravih kontrolnih ispitanika [(6,27±2,66) MN na 1000 BN stanica; (6,80±2,98) MN na 1000 BN stanica]. Prosječna se učestalost MN nije značajno razlikovala među skupinama kod kojih je došlo do spontanih pobačaja te među skupinama koje su imale normalne vrijednosti parametara kvalitete sjemena i onih koje nisu imale takve vrijednosti. Korelacije između niske koncentracije spermija (<20x106 mL-1), smanjene pokretljivosti spermija (<25 %), normalne morfologije (<30 %) i učestalosti MN bile su negativne, ali ne i statistički značajne. Utvrdili smo da je samo spol značajno utjecao na pojavnost MN u svih ispitanih bolesnika. Nije bilo značajnih razlika između dobnih skupina, kao ni između pušača i nepušača kod bolesnika i kontrolnih ispitanika. Zaključujemo da pojavnost MN u limfocitima bolesnika s poremećajima reproduktivnog sustava prati porast razine kromosomskih oštećenja koja nastaju kao posljedica složenih događaja koji uzrokuju poremećaje reproduktivnog sustava

Enhancement Of Dermal Fibroblast Isolation Method

ABSTRAC

Antitumor Effect of the Chalcone Analogue, (E) -1-(4-Ethoxy-3-Methoxyphenyl) -5- Methylhex-1-En-3-One on HeLa Cell Line

Chalcones represent precursor compounds for flavonoids biosynthesis in plants. Chalcones, 1,3-diaryl-2-propen-1-ones, have unique chemical structure with conjugated double bonds and delocalized π-electron system on both aromatic rings. Various studies have shown that chemical structure of chalcone is responsible for their antitumor effect. In our study, we have examined the antitumor effect of chalcone analogue (E) -1- (4-ethoxy-3-methoxyphenyl) -5-methylhex-1-en-3-one (CH) on HeLa cells. The antitumor efficiency of different CH concentrations was compared to the antitumor effects of dehydrozingerone and cisplatin. The viability of the cells was evaluated using MTT assay; type of the cell death was evaluated by Annexin V-FITC/7-AAD staining using FACS analysis; morphology changes of treated cells were visualized and compared to untreated cells using phase contrast microscopy. The result of our research showed that CH have a stronger antitumor compared to the effect both of dehydrozingerone and cisplatin. Our results indicated that chalcone analogue induced cell death via activation of apoptosis more powerfully compared to the apoptosis induced with dehydrozingerone and cisplatin

Induction of mitochondrial apoptotic pathway by raloxifene and estrogen in human endometrial stromal ThESC cell line

Introduction: Endometrial hyperplasia is a condition that occurs as a result of hormonal imbalance between estrogen and progesterone. Morphological disturbance of endometrial cells occurs consequently leading towards endometrial cancer. In therapy of endometrial hyperplasia SERMs are used to supress effects of locally high estrogen level in uterus. There is strong evidence suggesting that estrogen could be involved in cell death – apoptosis. There are no experimental data demstrating the direct apoptotic effect of both raloxifene and estrogen on the ThESC cell line. The aim of our study wa sto investigate both cytotoxic and apototic mechanism of raloxifene and estrogen – induced death in the ThESC cell line. Material and methods : In order to determine their cytotoxic and apoptotic effects, various doses of raloxifene and estrogen were applied to the ThESC cell line for 24h. After the treatment MTT assay, FACS analysis and immunofluoroscence method were conducted. Results : The results of this study for the first time demonstrated the cytotoxic and apoptotic effects of raloxifene and estrogen on human endometrial stromal cell line suggesting the involvement of the inner, mitochondrial apoptotic pathway. Conclusions : Our results demonstrated apoptotic effects of investigated drugs in the ThESC cell line through increasing the Bax/Bcl-2 ratio and activation of caspase 3

Older Hypertensive Patients’ Adherence to Healthy Lifestyle Behaviors

Non-pharmacological treatment including diet, body weight reduction, smoking cessation and physical activity, is very important part of hypertension treatment. The objective of this study was to investigate the adherence to healthy lifestyle behavior in the representative sample of the older hypertensive patients, and to investigate factors associated with adherence in the studied older population. The study was conducted on random sample of 362 long term hypertensive (> five years) patients older than 65 years of age, at Health Care Center of Kragujevac. Adherence was assessed using the structured questionnaire for the analysis of the implementation of both hypertension and diabetes guidelines in the primary care. Both bivariate and multivariate analyses were conducted. Nearly 35% of examined patients were highly adherent; they exercised regularly, avoided smoking for at least five years and consumed special healthy diet prescribed for hypertension. Another 35.6% of the cases reported exercising regularly, 39.5% followed the recommended diet for the hypertension, while 23.4% of the patients have still consumed cigarettes. Multivariate logistic regression demonstrated that received counseling on healthy lifestyle behaviors by physicians and lack of education predicted high adherence to healthy lifestyle behavior. In order to improve adherence of elderly hypertensive patients to healthy lifestyle, strengthening patient-physician relationships through efforts to enhance communication may be a promising strategy to enhance patients’ engagement in healthy lifestyle behaviors for hypertension. Such an improvement could be achieved through the education of both the physicians and patients

ANTITUMOR EFFECTS OF VANILLIN BASED CHALCONE ANALOGUES IN VITRO

Chalcones, as a large group of organic compounds, are widely implemented in various types of anti-cancer therapeutics. These plant metabolites are present in fruits, vegetables, spices, and have anti-tumor, anti-inflammation, immunomodulation, antibacterial and anti-oxidative activities, as well as many other pharmacological and biological effects. The aim of the present study was to investigate cytotoxic effects, type of cell death and mechanism of action of the newly synthesized vanillin based chalcone analogues, (CH1) and (CH2) on human colon cancer HCT-116 and noncancerous (control) MRC-5 cell lines. In order to compare effects of vanillin based chalcone analogues on investigated cell lines, as reference substances cisplatin (cisPt) and dehydrozingerone (DHZ) were used. Investigation of antitumor effect of chalcone analogues on HCT-116 cells was carried out by three methods MTT assay, flow cytometry and immunofluorescence analysis. The result of our investigation indicated that newly synthesized vanillin based chalcone analogues expressed powerful antitumor effect on cancer cells (HCT-116 cell line), while their effect on healthy cells (MRC-5 cell line) was not statistically significant. Vanillin based chalcone analogues caused overexpression and activation of mitochondrial Bax protein and caspase-3 in HCT-116 cells, indicating that their mechanism of antitumor action was mediated through activation of inner apoptotic pathway. These results indicate possible usefulness of CH1 and CH2 in antitumor therapy whether through its direct cytotoxic effect or as adjuvant therapy. Our results indicate possible usefulness of CH1 and CH2 vanillin based chalcone analogues in antitumor therapy

Association of SOD2 (Rs4880) and GPX1 (rs1050450) gene polymorphisms with risk of Balkan endemic nephropathy and its related tumors

© 2019 by the authors. Licensee MDPI, Basel, Switzerland. T. Background: Experimental data show that superoxide dismutase 2 (SOD2) is involved in ochratoxin (OTA)-induced nephrotoxicity, whereas clinical data indicate the role of SOD2 rs4880 or glutathione peroxidase 1 (GPX1) rs1050450 polymorphisms in end-stage renal disease and urothelial carcinoma risk, known to be the major complications of Balkan endemic nephropathy (BEN). Therefore, we hypothesized that SOD2 and GPX1 gene polymorphisms would influence the risk of BEN and its associated tumors. Materials and Methods: The study was conducted in 207 BEN patients and 86 controls from endemic areas. Results: Individuals with both copies of variant SOD2 allele, known for lower mitochondrial antioxidant protection, are at a significantly higher BEN risk (OR = 2.6, p = 0.021). No association was observed between GPX1 gene polymorphism and BEN risk. Combining SOD2 and GPX1 genotypes did not alter the risk of BEN development. Regarding the risk of urothelial tumors in BEN patients, none of the polymorphisms studied was significantly associated with the risk of these tumors. Conclusions: Polymorphism in SOD2 rs4880 gene affects the risk of BEN development. Hence, SOD2 genotyping could, together with a panel of other enzymes, be used as a biomarker of susceptibility in BEN areas