In order to make chemical characterization of two Origanum vulgare L. subspecies in Montenegro, the essential oils of five wild growing populations were analyzed. Among 67 oil constituents, in O. vulgare subsp. hirtum dominant one was oxygenated monoterpene carvacrol (74.3%), while in O. vulgare subsp. vulgare prevailed sesquiterpene hydrocarbons: germacrene D (15.4–27.9%) and β-caryophyllene (7.7–14.6%), and among oxygenated monoterpenes: α- terpineol (4.8–17.8%), linalyl acetate (0.5–9.6%), linalool (3.0–8.8%), thymol (0.2–8.3), terpinene 4-ol (1.5–8.3%). Several of the main essential oil constituents appeared to be highly intercorrelated. Strong positive correlations (r > 0.70; P < 0.01) were observed between α-terpineol and linalyl acetate, α-terpineol and thymol, linalyl acetate and thymol, γ-terpenene and carvacrol, (E)-β-ocimene and β-bisabolene, while strong negative correlations (r < −0.70; P < 0.01) were evidenced between γ-terpenene and β-caryophyllene, γ-terpenene and germacrene D, p-cymene and germacrene D. Multivariate analyses allowed the grouping of the populations into three distinct chemotypes. Population P5 (Origanum vulgare subsp. hirtum) was distinguished from O. vulgare subsp. vulgare populations by predominance of carvacrol, while within the typical subspecies the population P1 (Boljevići) separated from P2 (Radovče), P3 (Lipovo) and P4 (Grahovo) by high oil levels of α-terpineol, linalyl-acetate and thymol.

Three Schiff bases were synthesized by reaction of different benzaldehydes with amino acids. The characterization of these compounds was performed using IR spectroscopy, molecular calculations, thin-layer chromatography, determining the melting point and other physical characteristics. IR spectra for imino groups (C=N), which are characteristic of Schiff bases, show stretching frequency from 1629 to 1654 cm-1. The obtained spectral results were confirmed by molecular calculations using the density functional theory (DFT) and were performed before experimental work. The DFT global chemical reactivity descriptors were calculated and used to predict their relative stability and reactivity of synthesized compounds. The antimicrobial assay of all compounds were screened for Grampositive bacteria species: Staphylococcus aureus ATCC 25923; Methicillin-resistant Staphylococcus aureus: MRSA ATCC 33591; Bacillus subtilis ATCC 6633; and Enterococcus faecalis ATCC 29212, Gram-negative: Salmonella enterica ATCC 31194; Pseudomonas aeruginosa ATCC 9027; Escherichia coli ATCC 25922; Extended Spectrum Beta-Lactamase producing E. coli: ESBL E. coli ATCC 35218, and one yeast Candida albicans ATCC 1023.The highest values of inhibition zones were recorded for compound1, followed by the compound 3, while compound 2 performed inhibitory effect just in case of MRSA. DFT calculations show that antimicrobial activity has a good correlation with chemical reactivity descriptors obtained Schiff bases.

Authors : Alija Uzunovic, Sasa Pilipovic, Aida Sapcanin, Zahida Ademovic, Berina Pilipović Abstract : Capsaicin is a naturally occurring alkaloid extracted from capsicum fruit extracts of different of Capsicum species. It has been employed topically to treat many diseases such as rheumatoid arthritis, osteoarthritis, cancer pain and nerve pain in diabetes. The high degree of pre-systemic metabolism of intragastrical capsaicin and the short half-life of capsaicin by intravenous administration made topical application of capsaicin advantageous. In this study, we have evaluated differences in the dissolution characteristics of capsaicin patch 11 mg (purchased from market) at different dissolution rotation speed. The proposed patch area is 308 cm2 (22 cm x 14 cm; it contains 36 μg of capsaicin per square centimeter of adhesive). USP Apparatus 5 (Paddle Over Disc) is used for transdermal patch testing. The dissolution study was conducted using USP apparatus 5 (n=6), ERWEKA DT800 dissolution tester (paddle-type) with addition of a disc. The fabricated patch of 308 cm2 is to be cut into 9 cm2 was placed against a disc (delivery side up) retained with the stainless-steel screen and exposed to 500 mL of phosphate buffer solution pH 7.4. All dissolution studies were carried out at 32 ± 0.5 °C and different rotation speed (50± 5; 100± 5 and 150± 5 rpm). 5 ml aliquots of samples were withdrawn at various time intervals (1, 4, 8 and 12 hours) and replaced with 5 ml of dissolution medium. Withdrawn were appropriately diluted and analyzed by reversed-phase liquid chromatography (RP-LC). A Reversed Phase Liquid Chromatography (RP-LC) method has been developed, optimized and validated for the separation and quantitation of capsaicin in a transdermal patch. The method uses a ProntoSIL 120-3-C18 AQ 125 x 4,0 mm (3 μm) column maintained at 600C. The mobile phase consisted of acetonitrile: water (50:50 v/v), the flow rate of 0.9 mL/min, the injection volume 10 μL and the detection wavelength 222 nm. The used RP-LC method is simple, sensitive and accurate and can be applied for fast (total chromatographic run time was 4.0 minutes) and simultaneous analysis of capsaicin and dihydrocapsaicin in a transdermal patch. According to the results obtained in this study, we can conclude that the relative difference of dissolution rate of capsaicin after 12 hours was elevated by increase of dissolution rotation speed (100 rpm vs 50 rpm: 84.9± 11.3% and 150 rpm vs 100 rpm: 39.8± 8.3%). Although several apparatus and procedures (USP apparatus 5, 6, 7 and a paddle over extraction cell method) have been used to study in vitro release characteristics of transdermal patches, USP Apparatus 5 (Paddle Over Disc) could be considered as a discriminatory test. would be able to point out the differences in the dissolution rate of capsaicin at different rotation speed.

Bone is a specialized connective tissue that together with cartilage makes up the skeletal system. These tissues serve three functions: a) mechanical support and site of muscle attachment for locomotion; b) protective: for vital organs and bone marrow, and c) metabolic: reserve of ions for the entire organism, especially calcium and phosphate. It is also a composite structure, consisting of inorganic mineral crystals an extracellular organic matrix, cells, lipids and waters. The mineral crystals are analogous to the geologic mineral hydroxyapatite (Boskey and Coleman, 2010). The cells which produce, nurture and remodel the mineralized extracellular matrix, also respond to mechanical and other signals, which determine the properties (morphology and function) of the bone (Boskey and Coleman, 2010). Bone remodelling is a complex process which involves a number of cellular functions directed toward the coordinated resorption and formation of new bone (Ronchetti et al., 1996; Goldhaber, 1997; Jin et al., 2000). Catabolic agents (prostaglandine E2 (PGE2) and human parathyroid hormone (h-PTH) fraction 1-34) or anabolic agents (ascorbic acid (AA) and bone morphogenetic proteine 4 (BMP4) could stimulate bone resorption or bone formation directly in a bone organ-culture (Dempster et al., 1993; Goldhaber, 1997; Sampath Kuber, 1999). The main objective of the current research proposal is to establish calcium as an independent index for observing bone resorption and bone formation processes, which are age dependant.

Many studies highlight the health benefits of mushrooms, which are consequently becoming more and more interesting for researchers. The content of amino acids (AA), total phenolic content (TPC), and antioxidative activity (AAc) were determined in wild as well as cultivated mushrooms. The AA included: L-tryptophan (Trp), L-arginine (Arg), L-cysteine (Cys), L-methionine (Met), L-alanine (Ala), L-phenylalanine (Phe), L-lysine (Lys), L-valine (Val), L-glycine (Gly), and L-leucine (Leu). The wild mushrooms: Lactarius piperatus, Amanita caesarea, Lactarius deliciosus, Lycoperdon pyriforme, Macrolepiota procera, and cultivated mushrooms: Agaricus bisporus, Boletus aestivalis, Cantharellus cibarius, Pleurotus ostreatus, and Agaricus bisporus var. avellaneus were investigated in this study. The AA was determined by HPTLC method and quantified with a Camag TLC scanner and WINCAT software by scanning the plates at 540 nm. The TPC was spectrophotometrically estimated as gallic acid equivalents/g of fresh weight according to Folin-Ciocalteu’s method. The radical scavenging activity (RSA) of mushroom extracts was determined by DPPH assay. The highest content of Ala, Gly, Phe, Lys, Val, and Leu was found in Pleurotus ostreatus. The total phenolic content (mg GAE/g) in investigated mushrooms ranged from 1.90 to 35.56, and the % RSA ranged from 43.88 to 90.17. This study promotes the consumption of food rich in bioactive compounds, mushrooms being among such food. Further research on mushrooms from Bosnia and Herzegovina and their benefits in the overall maintenance of human health and protection from age-related diseases is necessary.