Phytochemical composition and antitumor activity of a new arctic lichen Anamylopsora pakistanica.

For the first-time, chemical composition and in vitro antitumor activity was investigated of a newly described lichen Anamylopsora pakistanica Usman & Khalid from the second highest plateau of the world (Deosai Plains, Pakistan). HPLC-UV method was used for identification of secondary metabolites and the acetone extract had higher values of TPC (41.90 mg GA/g) and TFC (75.37 mg RE/g) as compared to methanol extract. As chemical constituents 5,7-dihydroxy-6-methylphthalide, haematommic acid and alectorialic acid, were identified as major compounds. Atranol, alectorialin, gyrophoric acid and usnic acid were detected as minor substances. Acetone and methanol extracts induced a dose-dependent and time-dependent decrease in the viability of three types of tumour cells HeLa, HCT116 and MDA-MB-231. This lichen extract can induce S phase arrest in HeLa as compared to the untreated cells. Extract of this unique lichen, A. pakistanica, can be used safely as a significant source of biologically active compounds.

Antimicrobial treatment in invasive infections caused by Gordonia bronchialis: systematic review.

Background: Corynebacterium, Nocardia, Rhodococcus, Mycobacterium, as well as Gordonia genera belongs to the genus Gordonia, Actinomycetia class. Gordonia bronchialis is a nitrate-reducing, urease-producing, non-motile, force aerobe with a rod-like figure that is known to arrangement into sessile, cord-like groups. This systematic review aimed to establish whether and what invasive infections in humans were caused by Gordonia bronchialis, and to evaluate outcomes of administered antibiotic treatment. Methods: We have registered this systematic review in PROSPERO database of systematic reviews and meta-analyses with the number CRD42022369974. Results: A total of 24 publications were included (22 case reports and two case series) with 28 individual cases. The oldest patients had 92 years, and the youngest patients had 13 years. Clinical signs of infection were present in six patients (21%). All isolates were susceptible to ciprofloxacin, imipenem, and amikacin. Vancomycin was the most frequently used antibiotic with nine cases followed by ciprofloxacin, ceftriaxone, and amoxicillin/clavulanic acid. Conclusion: Although there are no standardized recommendations to date, successful treatment with a favorable outcome has most often been carried out with fluoroquinolones, vancomycin with or without aminoglycosides, as well as carbapenems.

Interleukin 33, soluble suppression of tumorigenicity 2, interleukin 27, and galectin 3 as predictors for outcome in patients admitted to intensive care units.

Intensive care units (ICUs) are expert hospital areas that provide treatment and 24 h care for people who are very sick. Sepsis represents a serious, severe condition and it can lead to septic shock and multiple organ dysfunction syndromes and is one of the most common reasons for patients' hospitalization in ICUs. We wanted to explore the prognostic values of interleukin (IL) 33, soluble suppression of tumorigenicity 2 (sST2), IL 27, and galectin 3 in critically-ill patients. We assumed that these parameters in combination or alone could predict mortality in ICU patients. This research represents a clinical non-randomized prospective study, performed at the Medical Military Academy, a tertiary care hospital in Belgrade, Serbia. The patients were divided in four groups: patients with sepsis (peritonitis, pancreatitis, trauma) and patients without sepsis (trauma). Total number of patients enrolled in the study was 151 and average years of patients were 56.48. The values greater than the cut-off were the predictors of mortality. The IL-33, IL-27 as well as galectin-3 can successfully predict the outcome of critically-ill patients in ICUs. The sST2, cannot predict death in critically-ill patients as a single prognostic factor. However, the combination of at least two biomarkers: IL-33, sST2, IL-27, and galectin-3, gives very significant results in predicting the outcome in patients admitted to ICUs.

Antimicrobial Treatment of Serratia marcescens Invasive Infections: Systematic Review.

Background: Serratia marcescens (SM) is a Gram-negative pathogen discovered by Italian pharmacist, Bizio, in 1819. According to the literature, S. marcescens is resistant to a wide range of antibiotics, including penicillin, cephalosporin, tetracycline, macrolide, nitrofurantoin, and colistin. We conducted a systematic review of published reports, determined what invasive infections could cause SM, and established the most appropriate antibiotic therapy. Methods: We registered this systematic review on the PROSPERO registry of systematic reviews-meta-analyses before we started our research (registration number CRD42022323159). The online searches of published studies were implemented via MEDLINE, the Cochrane Central Register of Controlled Trials, EBSCO, Scopus, Google Scholar, SCIndex, and the registry of clinical studies of human participants (ClinicalTrials.gov). Results: Our study included 32 published articles (9 case series and 23 case reports). There were 57 individual cases, respectively. The oldest patient was 97 years and the youngest patient was a newborn. S. marcescens was, in most cases, isolated from blood followed by urine and cerebrospinal fluid. In most cases, sensitivity was tested to cotrimoxazole (from 27 isolates, 10 showed resistance) followed by gentamicin (from 26 isolates, 3 showed resistance) as well as amikacin (from 21 isolates, none showed resistance). Patients died from an infection in 21 cases (31%). Conclusions: Treatment of SM infections should include carbapenems or aminoglycosides in combination with third-generation (and eventually fourth-generation) cephalosporin. Cotrimoxazole should be considered in cases of uncomplicated urinary infections.

Synthesis, Characterization, and Investigation of Anti-Inflammatory and Cytotoxic Activities of Novel Thiourea Derivatives of Naproxen.

The objective of this study was to synthesize seven novel thiourea derivatives of naproxen (8-14), examine the anti-inflammatory activity of the newly synthesized compounds, investigate the cytotoxic potential of both sets of synthesized compounds (1-7 and 8-14), and select the most promising anti-inflammatory and antitumor drug candidates. The results of the in vivo anti-inflammatory study clearly showed that compounds 8 and 9 were capable of decreasing paw edema, as evident from a high percentage of inhibition (44.83% and 49.29%, respectively). In addition, the results of in vitro enzyme inhibition assays demonstrated that neither of the newly synthesized compounds reached 50% inhibition of 5-LOX at concentrations lower than 100 µM. In terms of antitumor potential, derivatives 3 and 8 exhibited strong cytotoxic effects on the HeLa cell line, suggesting the involvement of the extrinsic pathway of apoptosis. According to the overall results obtained for both sets of synthesized molecules, derivatives 4 and 8 can be underlined as molecules with the strongest anti-inflammatory activity, while derivatives 3 and 8 are the most promising cytotoxic agents.

Validation of the fear of COVID-19 scale in a central Balkan country - Serbia.

Validation of the fear of introduction: High levels of fear of COVID-19 may be associated with increased levels of stress, anxiety, and depression, as well as decreased resilience and life expectancy. Objective: This study aimed to translate and confirm the Serbian version of the Fear of COVID-19 scale as well as to investigate its psychometric properties. Methods: The translation and intercultural adaptation of the Fear of COVID-19 scale was performed by the leading standard of the International Society for Pharmacoeconomics and Outcome Research. When the distribution was normal, the Kolmogorov-Smirnov test was used. The reliability of the Serbian version of FCV-19S was tested by measuring the internal consistency through the value of Cronbach's alpha. Results: The original version of the Fear of COVID-19 scale was tested on a sample of 256 subjects with a mean age of 25.38 ± 12.47. The Cronbach's alpha value was 0.864. We divided the scale by the split-half method (Spearman-Brown), and the value of the coefficient for the questionnaire as a whole was 0.882. Divergent criterion validity was tested through the non-parametric correlation between the scores of the Fear of COVID-19 scale and the Fear of Hospitalization scale. A score of the Fear of COVID-19 scale was calculated as the sum of each question for each of the respondents. Convergent criterion validity was tested through the non-parametric correlation between the scores of the Fear of COVID-19 scale and the Emotional Regulation Questionnaire. Conclusion: The validated version of the scale in Serbia complements versions available in other cultures and other languages and facilitates global studies related to mental health during the COVID-19 pandemic.

Newly synthesized palladium(II) complexes with aminothiazole derivatives: in vitro study of antimicrobial activity and antitumor activity on the human prostate cancer cell line.

Five new complexes of the palladium(II) ion (C1-C5) having the general formula [(PdL2)]Cl2 with some 2-aminothiazoles (L1-L5), where L1 = 2-amino-4-(3,4-difluorophenyl)thiazole, L2 = 2-amino-5-methyl-4-phenylthiazole, L3 = 2-amino-4-phenylthiazole, L4 = 2-amino-4-(4-chlorophenyl)thiazole, and L5 = 2-amino-4-(2,4-difluorophenyl)thiazole, have been synthesized and characterized by elemental microanalysis and infrared, 1H NMR and 13C NMR spectroscopy. The in vitro antimicrobial activity of the five ligands and the corresponding Pd(II) complexes is investigated. Testing is performed by the microdilution method and the minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) have been determined. Testing is conducted against 11 microorganisms (nine strains of pathogenic bacteria and two yeast species). The tested ligands and palladium(II) complexes show selective, high and moderate activity. There is a difference in antimicrobial activity between the ligands and the corresponding palladium(II) complexes. The complexes have significant anti-staphylococcal activity and activity on Pseudomonas aeruginosa which is better than the positive control. The interactions of newly synthesized palladium(II) complexes with calf thymus DNA (CT-DNA) were investigated using UV-Vis absorption and fluorescence spectroscopy. Analysis of UV-absorption and fluorescence spectra indicates the formation of a complex between the palladium(II) complexes and DNA. The high values of intrinsic binding constants, Kb, of the order 104 M-1 and Stern-Volmer quenching constants, KSV, of the order 105 M-1 indicated very good binding of all complexes to CT-DNA. Also, the new Pd(II) complexes show high cytotoxic activity towards the human prostate cancer cell line and insignificant activity towards non-cancerous human fibroblasts. Future research could additionally explore the biological activity of Pd(II) complexes presented in this paper and investigate the possibility of their implementation in clinical practice.

Antimicrobial treatment of Morganella morganii invasive infections: Systematic review.

BACKGROUND: Morganella morganii is a Gram-negative, rod-shaped, facultative anaerobic bacillus divided into two subspecies, morganii and sibonii. Previously classified as Proteus morganii, it belongs to human gut commensal microbiota. Nevertheless, on rare occasions, especially in nosocomial and postoperative environment as well as in patients with the impaired immune system and young children, it may cause potentially fatal systemic infection. OBJECTIVES: The aim of our systematic review was to determine whether and what invasive infections in humans were caused by Morganella morganii and to estimate outcomes of administered antibiotic management. DATA SOURCES: This systematic review was registered at the PROSPERO database of systematic reviews and meta-analyses before initiation of the research (registration number CRD42020171919). Study eligibility criteria and participants. patients of any age and both sex harbouring Morganella morganii as the only microorganism in bodily fluids or tissues, from where it was isolated and identified by one or more of the following diagnostic methods: conventional techniques including colony morphology, Vitek 2, API or BD Phoenix biochemical systems, as well as more sophisticated methods, such as Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and species-specific PCR for M. morganii. METHODS AND INTERVENTIONS: We have systematically searched MEDLINE, EBSCO, SCOPUS, SCINDEX and GOOGLE SCHOLAR for case reports and case series with M. morganii invasive infections. RESULTS: M. morganii can cause serious infections of different tissue in patients of any age. The most isolates were susceptible to ceftazidime, imipenem and amikacin. Majority of the patients completely recovered after antibiotic treatment. About 15% of the patients died despite of the therapy. Gentamicin was the most frequently used antibiotic in the treatment of infection caused by M. morganii. CONCLUSION: M. morganii invasive infections should be taken into consideration by the clinicians, especially in hospital conditions, due to its high degree of mortality and high potential of this bacterium to develop multidrug resistance. Treatment of M. morganii infections should include gentamycin in combination with third generation cephalosporin or another antibiotic to which M. morganii is susceptible (after testing isolates for third cephalosporin generation for the production of AmpC ß -lactamases).

Body Height of Elite Basketball Players: Do Taller Basketball Teams Rank Better at the FIBA World Cup?

Body height is among the most important attributes of basketball players. Whether it differs among the basketball players who compete at the world basketball championship (FIBA-WC) is unknown. The aim of this study was to investigate the differences between the basketball players from the teams ranked 1-16 and those ranked below 16th place. The body heights of all players from the last three FIBA-WCs were collected and allocated according to the ranking at the FIBA-WC and analyzed by position in team. An independent sample t-test was conducted to analyze the difference in body height of players ranked 1-16 and players who ranked below 16th place. The players from the first 16 teams were significantly taller at three positions: point guards (Difference = 2.92 cm, p < 0.05), shooting guards (Difference = 2.16 cm, p < 0.05), and small forwards (Difference = 2.49 cm, p < 0.01). Body height seems to be an important factor for the performance of the basketball team at the FIBA-WC. Considering that all players at the FIBA-WC went through rigorous selection process to be in their national teams, body height of the higher-ranked players could be used as a reference value.

Body composition characteristics measured by multichannel bioimpedance in young female basketball players: relation with match performance / Características de la composición corporal medidas por bioimpedancia multicanal en jugadoras de baloncesto jóvenes: relación con el rendimiento del partido

The aim of this research is to investigate the correlation of morphological characteristics, that is, body composition characteristics with the competitive performance of female basketball players of cadet (U16) age, in general and depending on the position played in the basketball team. The research involved female basketball players from the U16 section of the three best basketball clubs from the First Cadet League of Serbia (N = 30). This research consisted of measuring the characteristics of body composition, as a space of independent variables and the analysis of competitive activities where the dependent variable represents the level of achieved competitive performance. Based on the results of the Pearson Correlation Analysis, which was made for determining the degree of connection between the dependent variable and the independent variables, it can be seen that general level competitive performance is associated with the variables: percentage skeletal muscle mass, index of hypokinesy and protein-fat index. Viewed from the point of the position played in the team, the statistically significant relationship between the independent and the dependent variable was recorded only in the forwards position. Competitive and more successful forward players are distinguished by a small percentage of body fat mass, a higher percentage of muscle mass, a smaller hypokinesy index and a larger protein-fat index. The practical application of this research can be seen in the control system of applied training work, then selection, as well as in the creation of a model for morphological characteristics of players in the cadet (U16) age.

El objetivo del estudio fue investigar la correlación entre las características morfológicas, es especial, las características de la composición corporal y el rendimiento competitivo de jugadoras de baloncesto cadetes, en general y en cada posición en el equipo. La investigación incluyó a las jugadoras de baloncesto femenino U16 de los tres mejores clubes de Primera Liga de Cadetes de Serbia (N = 30). Esta investigación consistió en medir las características de la composición corporal, como un espacio de variables independientes, y el análisis de actividades competitivas, donde la variable dependiente representa el nivel de rendimiento competitivo alcanzado. Sobre la base de los resultados del análisis de coeficiente de correlación de Pearson, que se realizó con el propósito de determinar el grado de conexión entre la variable dependiente y las variables independientes, se pudo observar que, en general, el rendimiento competitivo está vinculado a las variables: porcentaje de masa muscular esquelética, índice de hipocinesia e índice proteínagrasa. Desde el punto de vista de posiciones dentro de un equipo de baloncesto, la relación estadísticamente importante entre la variable independiente y la variable dependiente ha sido registrada solamente en la posición de delantero. Las jugadoras delanteras más exitosas y competitivas se distinguen por un bajo porcentaje de grasa en su masa corporal, así como un mayor porcentaje de masa muscular, un índice de hipocinesia más bajo y un índice proteína-grasa más alto. El uso práctico de esta investigación se puede ver en el sistema de control del trabajo de entrenamiento aplicado, luego en la selección, así como en la creación de modelos de características morfológicas de jugadoras en la categoría cadetes (U16).

Synthesis, Anticancer Evaluation and Synergistic Effects with cisplatin of Novel Palladium Complexes: DNA, BSA Interactions and Molecular Docking Study.

BACKGROUND: In order to discover new agents for chemotherapy with improved properties compared to the existing agents and bearing in mind the fact that some Pd complexes possess better antitumor activity and exhibit less kidney toxicity compared to cisplatin, a series of novel square-planar palladium(II) complexes [Pd (L)2] (3a-f) with O,O bidentate ligands [L = ethyl 2- hydroxy-alkyl(aryl)-4-oxo-2-butenoate] were synthesized. METHODS: All complexes were characterized by spectral (UV-Vis, IR, NMR, ESI-MS) and X-ray analysis and examined for their cytotoxic effect on human cancer cell lines HeLa and MDA-MB 231 and normal fibroblasts (MRC-5). Fluorescence spectroscopic method was used for investigations of the interactions between CT-DNA or bovine serum albumin (BSA) and complex 3c. Viscosity measurements and molecular docking study were performed to confirm the mode of interactions between DNA and BSA and complex 3c. RESULTS: Complexes that showed the best results, 3c, 3d, and 3e, were placed under further investigations. Selected complexes induced apoptosis and cell cycle arrest in HeLa and MDA-MB 231 cells. Low concentrations of 3c and 3e showed strong to moderate synergism with low concentrations of cisplatin. The interaction of 3d with cisplatin was antagonistic in all used concentrations, but low IC50 value indicates its usefulness as a single cytotoxic agent. It was also noted that the change of viscosity is more pronounced in DNA solution after addition of complex 3c. CONCLUSION: Obtained results indicate that the novel palladium(II) complexes have the potential to become candidates for treatment in anticancer therapy.

Naphthoquinone rich Onosma visianii Clem (Boraginaceae) root extracts induce apoptosis and cell cycle arrest in HCT-116 and MDA-MB-231 cancer cell lines.

In the present study, five root extracts of Onosma visianii Clem were investigated for their in vitro cytotoxic activity. On the basis of HPLC-PDA analysis, these extracts have proved to be a rich source of naphthoquinones as natural colourants for food and cosmetic industry. All investigated root extracts contain acetylshikonin, isobutyrylshikonin and α-methylbutyrylshikonin as major compounds. As the most abundant source of active compounds for antitumour therapy, acetone, chloroform and ethyl acetate extracts showed strong cytotoxic activity towards HCT-116 and MDA-MB-231 cancer cell lines. Also, these extracts induced apoptosis and cell cycle arrest in HCT-116 and MDA-MB-231 cancer cell lines.

Enhanced cytotoxicity and apoptosis by raloxifene in combination with estrogen and methotrexate in human endometrial stromal cells.

Endometrial hyperplasia is a condition that may lead to the development of endometrial carcinoma. Initially, changes of the endometrium are caused by the estrogen's hyperstimulation that may lead to the development of an irregular bleeding and the infertility problems. Therapy of endometrial hyperplasia is limited to medical and surgical approaches. During the past decade, the new types of drugs were developed for the treatment of the endometrial hyperplasia. Here, for the first time, we investigated the cytotoxic effects of the various combinations of estrogen, raloxifene, and methotrexate in human ThESC cell line as a possible potential treatment of the endometrial hyperplasia. Our aim was to investigate and to determine the most efficient combination of investigated drugs in ThESC cells during 24-hr period using MTT assay, FACS analysis, and immunofluorescence staining. Our results demonstrated that the combination of raloxifene with methotrexate efficiently induced both the cytotoxicity and apoptosis in ThESC cells when compared to their single effect, as well as to the effect of combined treatment of raloxifene with estrogen. The application of the low doses of methotrexate combined with raloxifene offers all advantages of a potential beneficial antitumor match in cancer chemoprevention and therapy.

Antibacterial and cytotoxic activities of naphthoquinone pigments from Onosma visianii Clem.

In this study, the antibacterial and cytotoxic activities of isolated compounds from the roots of Onosma visianii were investigated. By using different chromatographic techniques and appropriate spectroscopic methods, the seven naphthoquinones were described: deoxyshikonin ( 1 ), isobutyrylshikonin ( 2 ), α-methylbutyrylshikonin ( 3 ), acetylshikonin ( 4 ), ß-hydroxyisovalerylshikonin ( 5 ), 5,8-O-dimethyl isobutyrylshikonin ( 6 ) and 5,8-O-dimethyl deoxyshikonin ( 7 ). Among the tested compounds, 3 and 4 exhibited the highest antibacterial activities toward all tested bacterial species (MIC50 and MIC90 for gram positive bacteria: 6.40 µg/mL-12.79 µg/mL and 6.82 µg/mL-13.60 µg/mL, respectively; for gram negative bacteria: 4.27 µg/mL-8.53 µg/mL and 4.77 µg/mL-9.54 µg/mL, respectively). Also, naphthoquinones 3 and 4 exhibited strong cytotoxic activity against MDA-MB-231 cells (IC50 values 86.0 µg/mL and 80.2 µg/mL, respectively), while compounds 1 , 3 , 4 and 5 significantly decreased viability of HCT116 cells (IC50 values of 97.8 µg/mL, 15.2 µg/mL, 24.6 µg/mL and 30.9 µg/mL, respectively). Our results indicated that all tested naphthoquinone pigments are potential candidates for clinical uses as antibacterial and cytotoxic agents.

Cytokine profile in chronic hepatitis C: An observation.

The data addressing cytokine profile in chronically infected HCV patients are conflicting, ranging from Th1 or Th2 cytokine prevalence to the expression of both types of cytokines. Therefore, the aim of this study was to evaluate cytokine profile in these patients. Cytokine sera levels in HCV patients and healthy controls were evaluated using 13plex FlowCytomix Multiplex. Median values of both proinflammatory and anti-inflammatory cytokines were lower in HCV patients then in controls. In addition, the number of subjects producing detectable quantities of cytokines was significantly lower in the group of HCV patients. Yet, cytokine levels in those patients were remarkably heterogeneous ranging from low to extremely high, much higher than the maximal values in control group. Similarly, grouping data according to HCV genotype, HCV RNA load, ALT/AST ratio and the stage of fibrosis showed marked standard deviations, reflecting high intragroup diversity. No correlation was found between each disease-related factor and cytokine levels. Patients investigated in our and similar studies were disparate pursuant to characteristics of the hosts, pathogen and course of the disease. Therefore, the inconsistency of the literature data regarding cytokine pattern in chronic HCV patients may be a consequence of the disregarded/overlooked heterogeneity of these patients.

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 24 h. 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.

Synthesis, characterization, biological activity, DNA and BSA binding study: novel copper(ii) complexes with 2-hydroxy-4-aryl-4-oxo-2-butenoate.

A serie of novel square pyramidal copper(ii) complexes [Cu(L)2H2O] (3a-d) with O,O-bidentate ligands [L = ethyl-2-hydroxy-4-aryl-4-oxo-2-butenoate; aryl = 3-methoxyphenyl-2a, (E)-2-phenylvinyl-2b, (E)-2-(4'-hydroxy-3'-methoxyphenyl)vinyl-2c, 3-nitrophenyl-2d, 2-thienyl-2e] were synthesized and characterized by spectral (UV-Vis, IR, ESI-MS and EPR), elemental and X-ray analysis. The antimicrobial activity was estimated by the determination of the minimal inhibitory concentration (MIC) using the broth microdilution method. The most active antibacterial compounds were 3c and 3d, while the best antifungal activity was showed by complexes 3b and 3e. The lowest MIC value (0.048 mg mL-1) was measured for 3c against Proteus mirabilis. The cytotoxic activity was tested using the MTT method on human epithelial carcinoma HeLa cells, human lung carcinoma A549 cells and human colon carcinoma LS174 cells. All complexes showed extremely better cytotoxic activity compared to cisplatin at all tested concentrations. Compound 3d expressed the best activity against all tested cell lines with IC50 values ranging from 7.45 to 7.91 µg mL-1. The type of cell death and the impact on the cell cycle for 3d and 3e were evaluated by flow cytometry. Both compounds induced apoptosis and S phase cell cycle arrest. The interactions between selected complexes (3d and 3e) and CT-DNA or bovine serum albumin (BSA) were investigated by the fluorescence spectroscopic method. Competitive experiments with ethidium bromide (EB) indicated that 3d and 3e have a propensity to displace EB from the EB-DNA complex through intercalation suggesting strong competition with EB [Ksv = (1.4 ± 0.2) and (2.9 ± 0.1) × 104 M-1, respectively]. Ksv values indicate that these complexes bind to DNA covalently and non-covalently. The achieved results in the fluorescence titration of BSA with 3d and 3e [Ka = (2.9 ± 0.2) × 106 and (2.5 ± 0.2) × 105 M, respectively] showed that the fluorescence quenching of BSA is a result of the formation of the 3d- and 3e-BSA complexes. The obtained Ka values are high enough to ensure that a significant amount of 3d and 3e gets transported and distributed through the cells.

Chelidonium majus crude extract inhibits migration and induces cell cycle arrest and apoptosis in tumor cell lines.

ETHNOPHARMACOLOGICAL RELEVANCE: Chelidonium majus L (Papaveraceae) is widely used in alternative medicine for treatment of various disorders. Antitumor activities of alkaloids isolated from this plant have been reviewed, while there are only a few studies that examine properties of the whole extract. AIM OF THE STUDY: The aim of the present study was to investigate direct cytotoxic effects, as well as indirect antitumor effects of Chelidonium majus ethanolic extract against different tumor cell lines,. MATERIALS AND METHODS: MTT and SRB assays were performed to estimate cytotoxic effects of Chelidonium majus extract against human tumor cell lines A549, H460, HCT 116, SW480, MDA-MB 231 and MCF-7 and peripheral blood mononuclear cells from healthy individuals. Cell cycle analysis was performed by flow cytometry. Type of cell death induced by extract was determined by flow cytometry and cell morphology assessment. Inhibitory effect on migration of cancer cells was assessed by wound healing assay. RESULTS: Chelidonium majus extract showed selective time- and dose-dependent increase of cytotoxicity in all six cell lines, with individual cell line sensitivities. Extract promoted cell cycle arrest and induced apoptosis. Cotreatment with doxorubicin enhanced cytotoxicity of the drug. Also, inhibitory effect on migration was shown with non-toxic extract concentration. CONCLUSIONS: These results indicate possible usefulness of Chelidonium majus crude extract in antitumor therapy, whether through its direct cytotoxic effect, by prevention of metastasis, or as adjuvant therapy.

Effects of cardiovascular drugs on TSH serum levels in patients on replacement therapy after thyroidectomy.

OBJECTIVE: Hypothyroidism is one of most common endocrine disorders resulting from deficiency of thyroid hormones. The aim of our study was to investigate whether cardiovascular drugs as well as gender, age, body-mass index, and habits, like smoking or drinking coffee affect thyroid-stimulating hormone (TSH) level in hypothyroid patients with thyroxine replacement therapy who suffer from cardiovascular disease. MATERIALS: The study was conducted on 150 hypothyroid patients who underwent total thyroidectomy for benign reasons; they were divided into five treatment groups: levothyroxine only group and, according to the drugs they had in therapy alongside levothyroxine, the angiotensinconverting enzyme inhibitors group, the selective ß-blockers group, the calcium antagonists group, as well as the nitrates group. A retrospective cohort study was conducted in the Clinical Center Kragujevac, Serbia, during the period of January 2012 to October 2014. All patients' data were collected both from participants' health records and questionnaires that patients completed, including data about habits, like smoking or drinking coffee. RESULTS: TSH values were significantly higher in the group of patients with selective ß-blockers in therapy alongside levothyroxine, compared to all the other study groups. The values of TSH level did not significantly differ among the other therapy groups. On the other hand, cigarette smoking was a risk factor that decreased TSH levels in patients on thyroid replacement therapy. CONCLUSIONS: Our study shows that selective ß-1 blockers can increase, while cigarette smoking can decrease TSH serum levels in hypothyroid patients on thyroid-replacement therapy.

Chrysin induces apoptosis in peripheral blood lymphocytes isolated from human chronic lymphocytic leukemia.

Chronic lymphocytic leukemia (CLL) develops due to an imbalance between apoptosis and proliferation of B lymphocytes. Chrysin induced apoptosis in leukemia cell lines such as U937, MO7e, THP-1 and HL-60, but there has not yet been data demonstrating the apoptotic effect of chrysin on CLL cells. Therefore, in our investigation we examined the cytotoxicity of chrysin against two leukemia cell lines, MOLT-4 and JVM-13, peripheral blood lymphocytes isolated from B-CLL patients and peripheral blood mononuclear cells (PBMC) from healthy individuals in vitro. The effect of chrysin on viability of MOLT-4 and JVM-13 cell lines, B-CLL cells derived from 28 patients and PBMC from 16 healthy subjects was determined by MTT assay. The type of cell death induced by chrysin was verified by Annexin V/7AAD assay and acridine orange and ethidium bromide (AO/EB) staining assay. Intracellular localisation and endogenic expression of apoptotic proteins including Bax, Bcl-2, cytochrome c and caspase-3 were determined by flow cytometry and fluorescent microscopy. Our results demonstrated that exposure of MOLT-4, JVM-13 cell lines and B-CLL cells to the concentration of chrysin of 10µM and higher selectively decreased viability of cells in this cell population, but not in the PBMC derived from healthy subjects; LC50 values of chrysin for B-CLL cells were 51µM for 24 hours and 32µM for 48 hours of incubation, respectively. Our findings demonstrated that chrysin induces the activation of proapoptotic Bax and decreases the expression of antiapoptotic Bcl-2 protein, releases cytochrome c from mitochondria into cytosol and cleavages/activates caspase-3, subsequently leading to the activation of apoptosis of B-CLL cells. Together, these findings suggest that chrysin selectively induces apoptosis of peripheral blood lymphocytes isolated from human chronic lymphocytic leukemia patients via mitochondrial pathway in vitro and that it might have a promising role as a potential future antileukemic remedy.