Rutin is quercetin heteroside widely present in plants that exhibit many health-beneficial effects, such as strengthening the capillary walls, reducing the harmful effects of LDL cholesterol, and reducing the risk of cardiovascular diseases. Rutin was determined from three <i>Allium</i> species (<i>A. cepa</i>, <i>A. sativum</i>, and <i>A. ursinum</i>) collected in Bosnia and Herzegovina by the HPLC method associated with electrochemical detection. The analysis was performed from methanol extracts of bulb and leaf of garlic, bulb and leaf of onion, and leaves of wood garlic. Rutin was present in all of the examined samples. The highest rutin content was found in garlic leaves (0.78 ± 0.09 mg g^–1), and the lowest in onion bulbs (0.04 ± 0.10 mg g^–1). The contents of rutin were higher in leaf samples, suggesting that leaves of onion and garlic are recommended as a better natural source of this glycoside.

Black cumin (Nigella sativa L. [N.sativa]) seed extracts demonstrated numerous beneficial biological effects including, among others, antidiabetic, anticancer, immunomodulatory, antimicrobial, anti-inflammatory, antihypertensive, and antioxidant activity. To better understand the phytochemical composition of N. sativa seeds, methanol seed extracts were analyzed for phenolic acid and flavonoid content. Furthermore, we tested N. sativa methanol, n-hexane, and aqueous seed extracts for their inhibitory activity against butyrylcholinesterase (BChE) and catalase (CAT) as well as for antimicrobial activity against several bacterial and a yeast strains. The phenolic content of N. sativa was analyzed using ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). The inhibition of BChE was assessed by modified Ellman's method, and the inhibition of CAT was determined by monitoring hydrogen peroxide consumption. The extracts were tested against Bacillus subtilis, Staphylococcus aureus, Salmonella enterica, and Escherichia coli using the agar diffusion method. The UHPLC-MS/MS method allowed the identification and quantification of 23 phenolic compounds within 15 minutes. The major components found in N. sativa seed extract were sinapinic acid (7.22 ± 0.73 µg/mg) as a phenolic acid and kaempferol (11.74 ± 0.92 µg/mg) as a flavonoid. All extracts showed inhibitory activity against BChE, with methanol seed extract demonstrating the highest inhibitory activity (inhibitory concentration 50% [IC50] 79.11 ± 6.06 µg/ml). The methanol seed extract also showed strong inhibitory activity against CAT with an IC50 value of 6.61 ± 0.27 µg/ml. Finally, the methanol extract exhibited considerable inhibitory activity against the tested microbial strains. Overall, this is the first study to investigate the ability of black cumin seed extracts to inhibit CAT. Our results indicate that N. sativa seed can be considered as an effective inhibitor of CAT activity.

Summary Background: The production of erythrocytes is regulated by the hormone erythropoietin (EPO), which maintains the blood haemoglobin (Hb) levels constant under normal conditions. Human EPO is a glycoprotein hormone and its synthesis is controlled by the hypoxia-inducible transcription factor. The aim of this study was to establish EPO and Hb levels in patients with chronic kidney disease (CKD), as well as in control subjects, and to investigate the relationship between these parameters. Methods: This cross-sectional, observational study included 356 subjects with CKD divided into 4 subgroups according to their glomerular filtration rate (GFR). The control group consisted of 206 age and sex matched healthy subjects with GFR rate ≥90 mL/min/1.73 m2. EPO, Hb and serum creatinine levels were determined by using immunochemical and spectrophotometric methods. GFR was determined using the MDRD formula. Results: The CKD patients had significantly lower levels of haemoglobin (p<0.0005) and hematocrit (p<0.0005) compared to control group. Our results showed that Hb levels decreased, whereas serum creatinine increased with the increasing renal failure. The CKD patients in all four groups had significantly lower (p<0.0005) Hb levels, and significantly higher (p<0.0005) creatinine levels compared to the control group. The median EPO in group I and II were significantly higher (p=0.002; p=0.018), while median EPO in group III and IV were significantly lower (p=0.03; p=0.011) compared to the control group. Conclusions: In patients with CKD, GFR positively correlated with Hb and EPO, while the correlation between GFR and serum creatinine was negative.

Ephedra is a plant with a long history of medicinal use, the main indications being asthma, rhinitis and bronchospasm. However, nowadays it is often misused/abused for weight-loss, performance enhancing and doping. The pharamcological effects are ascribed to the ephedrine alkaloids present in the plant. The aim of the study was to quantify Norephedrine (NE), Ephedrine (E), Pseudoephedrine (PE) and Methylephedrine (ME) in herb extracts of nine different Ephedra species using high performance liquid chromatography with UV detection (HPLC-UV). The alkaloids were separated on the column Symmetry C18 (5 µm, 3.9 x 150 mm). The mobile phase was delivered at the flow rate of 1 ml/min. The mobile phase was prepared with 390 ml of acetonitrile, 610 ml of pure water, 0.8 ml of o-phosphoric acid and 4.8 g of sodium lauryl sulfate. The UV detector operated at 208 nm. We found that the utilized HPLC-UV method for separation and quantification of ephedrine alkaloids was very selective, sensitive and simple. The total content of NE, E, PE and ME ranged from 0.4 mg/g dry weight (Ephedra foeminea, from Austria) to 71.9 mg/g dry weight (Ephedra major, from Morocco). None of the investigated alkaloids were detected in Ephedra altissima.

Rutin is quercetin heteroside widely present in plants that exhibit many health-beneficial effects, such as strengthening the capillary walls, reducing the harmful effects of LDL cholesterol, and reducing the risk of cardiovascular diseases. Rutin was determined from three Allium species (A. cepa, A. sativum, and A. ursinum) collected in Bosnia and Herzegovina by the HPLC method associated with electrochemical detection. The analysis was performed from methanol extracts of bulb and leaf of garlic, bulb and leaf of onion, and leaves of wood garlic. Rutin was present in all of the examined samples. The highest rutin content was found in garlic leaves (0.78 ± 0.09 mg g−1), and the lowest in onion bulbs (0.04 ± 0.10 mg g−1). The contents of rutin were higher in leaf samples, suggesting that leaves of onion and garlic are recommended as a better natural source of this glycoside.

The medicinal significance of Ephedra is based on the sympathomimetic properties of ephedrine (E) alkaloids. Pharmacological effects depend on the phytocomposition of individual Ephedra species. The aim of this study was to measure the total alkaloids content (TAC), total phenolics content (TPC), and total flavonoids content (TFC) and determine their relationship in dry herb of Ephedra major, Ephedra distachya subsp. helvetica, Ephedra monosperma, Ephedra fragilis, Ephedra foeminea, Ephedra alata, Ephedra altissima and Ephedra foliata. Nowadays, medicinal use of Ephedrae herba is limited, but the abuse of its psychostimulants is rising. In this study, TAC, TPC and TFC were determined using spectrophotometric methods. For the first time, ultra-performance liquid chromatography with ultraviolet detection (UPLC-UV) was used for separation and quantification of E-type alkaloids of various Ephedra species. The highest TPC and TFC were found in E. alata (53.3 ± 0.1 mg Gallic acid equivalents/g dry weight, 2.8 mg quercetin equivalents/g dry weight, respectively). The total content of E and pseudoephedrine determined by UPLC-UV varied between 20.8 mg/g dry weight (E. distachya subsp. helvetica) and 34.7 mg/g dry weight (E. monosperma). The variable content and ratio between secondary metabolites determined in different Ephedra species reflects their metabolic activities. Utilization of UPLC-UV unveiled that this technique is sensitive, selective, and useful for separation and quantification of different alkaloids in complex biological matrixes. The limit of detection was 5 ng. Application of UPLC-UV can be recommended in quick analyses of E-type alkaloids in forensic medicine and quality control of pharmaceutical preparations.

Introduction: Hypoxia is a basic stimulant in production of erythropoietin (EPO). The primary function of erythrocytes is the transport of oxygen to tissues. Erythropoietin stimulates erythropoiesis which leads to increased production of erythrocytes- their total mass. This increases the capacity of the blood to carry oxygen, reduces the hypoxic stimulus and provides a negative feedback of stopping EPO production. The aim of this study was to establish a quantitative relationship between the concentration of erythropoietin, hemoglobin and hematocrit in different values of renal insufficiency. Material and methods: The survey was conducted on 562 subjects divided into two groups: with and without renal insufficiency. EPO, hemoglobin, hematocrit, serum creatinine and additional parameters iron, vitamin B12, and folic acid were determined by using immunochemical and spectrophotometric methods and glomerular filtration rate (GFR) was calculated as well. Results: EPO values (median) grow to the first degree of renal insufficiency, as compared to EPO values of healthy subjects, this increase is statistically significant, p=0.002. With further deterioration of renal function the values of EPO between all pathological groups are decreasing, and this decrease is statistically significant between first and second degree of renal insufficiency (RI) p<0.001. In the group of healthy subjects EPO is correlated rho = -0.532, p <0.0005 with hematocrit. The correlations are negative and strong and can be predicted by regression line (EP0 = 41.375- Hct * .649; EPO = 61.41–Hb * 0.355). In the group of subjects with the first degree of renal insufficiency EPO is in correlation with hematocrit rho=-0.574, p<0, 0005. It is also correlated with hemoglobin rho=-0.580, p< 0.0005. The correlation is negative (EP0= 42.168- Hct * 0.678). In the group of subjects with the third degree of renal insufficiency EPO is in correlation with hemoglobin rho=0.257, p=0.028. The correlation is medium strong and positive. In the group of subjects with third and fourth degree of renal insufficiency EPO is not in correlation with hemoglobin and hematocrit p>0.05. Conclusion: Renal dysfunction, depending on the level of RI effects differently on the biosynthesis of EPO in a diseased kidney, and consequently it also has a different effect on biosynthesis of HB in bone marrow and its content in the blood.

Platelet hyperactivity and platelet interaction with endothelial cells contribute to the development and progression of many cardiovascular diseases such as atherosclerosis and thrombosis. The impact of platelet activity with different pharmacological agents, such as acetylsalicylic acid and coumarin derivatives, has been shown to be effective in the prevention of cardiovascular disease. Artemisia dracunculus, L. Asteraceae (Tarragon) is used for centuries in the daily diet in many Middle Eastern countries, and it is well known for its anticoagulant activity. The present study investigates the presence of coumarins in tarragon leaves and subsequently determines the extract with a major amount of coumarin derivatives. The solvents of different polarities and different pH values were used for the purpose of purifying the primary extract in order to obtain fractions with the highest coumarin content. Those extracts and fractions were investigated for their anticoagulant activity by determining prothrombin time (PT) and the international normalized ratio (INR), expressed in relation to the coagulation time of the healthy person. Purified extracts and fractions obtained from plant residue after essential oil distillation, concentrated in coumarin derivatives, showed the best anticoagulant activity, using samples of human blood. INR maximum value (2.34) and consequently the best anticoagulant activity showed the methanol extract at concentration of 5%.  The INR value of normal plasma in testing this extract was 1.05.

Aims: Study investigates relation of sulfur compounds content in garlic and different type of manures that were used for rearing of this vegetable. Measured parameter was total sulfur which could be a valid parameter of content of aromatic and medicinal sulfur compounds in garlic (alliin etc.). Study Design: Quality of garlic mainly depends from content of sulfur compounds, there are articles related to influence of mineral fertilizer on the quality of garlic bulbs but effect of manures hasn’t been enough investigated. This research should give an estimation of effect of some manures on the quantity of sulfur compounds in garlic.