Bioassay-guided fractionation of pterocarpans from roots of Harpalyce brasiliana Benth

Pterocarpans, a special kind of isoflavonoids possessing two contiguous benzofuran and benzopyran rings, have been reported as possessing several biological activities. In order to isolate and identify the active principles possibly responsible for the stronger activity of the EtOH extract from roots of Harpalyce brasiliana on the antimitotic assay using sea urchin egg development, a bioassay-guided fractionation was performed. Six bioactive pterocarpan derivatives: 4'-dehydroxycabenegrin A-I, leiocarpin, medicarpin, cabenegrins A-I and A-II, and maackiain were isolated from the chloroform fraction of H. brasiliana extract. Leiocarpin was the most active on the sea urchin egg assay with IC(50) values ranging from 0.1 to 1.2 microg/mL, followed by 4'-dehydroxycabenegrin A-I. The isolated compounds were also tested for cytotoxicity against tumor cell lines in cultures, where 4'-dehydroxycabenegrin A-I was the most active, followed by leiocarpin. Additionally, some studies on the structure-activity relationship of these pterocarpans are suggested.     

Antioxidant capacity of seminal plasma measured by TAS Randox

BACKGROUND: An imbalance between reactive oxygen species (ROS) generation in sperm and antioxidant capacity of seminal plasma has been linked to male infertility. The antioxidant power of biological fluids can be evaluated either by measurement of individual antioxidants or total antioxidant capacity (TAC). The aim of this study was to assess whether TAS Randox can also be used for seminal plasma antioxidant capacity estimation. MATERIAL AND METHODS: Freshly thawed aliquots of seminal plasma and blood serum of 38 males from infertile couples and 24 healthy normospermic controls were simultaneously assayed using TAS Randox reagents on the Hitachi Modular P800 Analyzer. Semen analysis was performed according to WHO guidelines. ROS in fresh sperm suspension in phosphate buffered saline was measured by chemiluminescence immediately after separation of seminal plasma. RESULTS: Semen analysis showed that in our study group only 14 males were normospermic and 24 males had mostly combined pathologies. The medians for ROS production were similar in both the study and the control groups (4850 and 5450 RLU/min, resp). Seminal plasma TAS levels were significantly lower (p<0.02) in the study group while blood serum TAS levels were similar in both groups. A significant positive correlation (p<0.05) between TAS in seminal plasma and serum was found, seminal plasma levels being on average 1.4 times higher. CONCLUSIONS: The TAS Randox kit may be used for clinical studies intended to identify decreased antioxidant power in the seminal fluid of infertile men.     

Distinct faces of the Ku heterodimer mediate DNA repair and telomeric functions

The Ku heterodimer, comprised of Ku70 and Ku80 subunits, is a conserved complex involved in nonhomologous end-joining (NHEJ). However, it also functions in maintenance of telomeres, chromosome termini normally resistant to end-joining events. To elucidate the spatial organization of these functions, we rationally guided Ku mutagenesis in yeast with real-valued evolutionary trace (rvET). This revealed two ancestrally related alpha-helices: one on the Ku70 surface that is required in yeast for NHEJ, and a second on the Ku80 surface that is required in yeast for telomeric heterochromatin formation. When bound to a DNA end, the surface containing the NHEJ-specific Ku70 helix is oriented toward the DNA terminus, whereas the surface containing the telomeric function-specific Ku80 helix faces inward, toward telomeric chromatin, when bound to a telomere. We propose a 'two-face' model for Ku and that divergent evolution of these faces allowed Ku's dual role in NHEJ and telomere maintenance.     

Participation of a Specific Substrate Degrading Strain in a Mixed Bacteria Culture as a Result of Ciliate Grazing

Participation of nitrilotriacetic acid degrading bacterial strain NTA-1 in the continuous-cultivated mixed culture was studied under different conditions including predation pressure of the ciliate Dexiostoma campyla (STOKES , 1886). From the viewpoint of dispersed/flocculated biomass distribution, significant relationships between NTA-1 and total bacteria ratio, and dispersed and total biomass ratio were proved in the systems without high concentrations of ciliates. The ciliate concentrations reaching 10⁴ ml⁻¹ stabilized flocculated biomass growth without directly affecting NTA-1 portion. Using fluorescently labelled NTA-1 bacteria, filter feeding rates of ciliates were evaluated (maximum individual uptake rate upon NTA-1 bacteria as a number of bacteria per ciliate per hour being 120 h⁻¹ and 260 h⁻¹ under ciliate division rate of 0.3 day⁻¹ and 1 day⁻¹, respectively). Biomass balance showed that dispersed NTA-1 bacteria should not serve as the sole feeding source for these free-swimming ciliates. The role of diversity of mixed bacterial diet in ciliate growth and the role of ciliate predation in stabilizing bacterial assemblage structure was proved.     

Poly(ADP-ribose) Glycohydrolase Is Present and Active in Mammalian Cells as a 110-kDa Protein

Poly(ADP-ribose) glycohydrolase (PARG) is the major enzyme responsible for the catabolism of poly(ADP-ribose), a reversible covalent-modifier of chromosomal proteins. Purification of PARG from many tissues revealed heterogeneity in activity and structure of this enzyme. To investigate PARG structure and localization, we developed a highly sensitive one-dimensional zymogram allowing us to analyze PARG activity in crude extracts of Cos-7, Jurkat, HL-60, and Molt-3 cells. In all extracts, a single PARG activity band corresponding to a protein of about 110 kDa was detected. This 110-kDa PARG activity was found mainly in cytoplasmic rather than in nuclear extracts of Cos-7 cells.     

Cytokinins in Tobacco and Wheat Chloroplasts. Occurrence and Changes Due to Light/Dark Treatment

Although cytokinins (CKs) affect a number of processes connected with chloroplasts, it has never been rigorously proven that chloroplasts contain CKs. We isolated intact chloroplasts from tobacco (Nicotiana tabacum L. cv SR1) and wheat (Triticum aestivum L. cv Ritmo) leaves and determined their CKs by liquid chromatography/tandem mass spectroscopy. Chloroplasts from both species contained a whole spectrum of CKs, including free bases (zeatin and isopentenyladenine), ribosides (zeatin riboside, and isopentenyladenosine), ribotides (isopentenyladenosine-5'-monophosphate, zeatin riboside-5'-monophosphate, and dihydrozeatin riboside-5'-monophosphate), and N-glucosides (zeatin-N(9)-glucoside, dihydrozeatin-N(9)-glucoside, zeatin-N(7)-glucoside, and isopentenyladenine-N-glucosides). In chloroplasts there was a moderately higher relative amount of bases, ribosides, and ribotides than in leaves, and a significantly increased level of N(9)-glucosides of zeatin and dihydrozeatin. Tobacco and wheat chloroplasts were prepared from leaves at the end of either a dark or light period. After a dark period, chloroplasts accumulated more CKs than after a light period. The differences were moderate for free bases and ribosides, but highly significant for glucosides. Tobacco chloroplasts from dark-treated leaves contained zeatin riboside-O-glucoside and dihydrozeatin riboside-O-glucoside, as well as a relatively high CK oxidase activity. These data show that chloroplasts contain a whole spectrum of CKs and the enzymatic activity necessary for their metabolism.     

Methods for measuring acoustic power of an ultrasonic neurosurgical device

Measurement of the acoustic power in high-energy ultrasonic devices is complex due to occurrence of the strong cavitation in front of the sonotrode tip. In our research we used three methods for characterization of our new ultrasonic probe for neuroendoscopic procedures. The first method is based on the electromechanical characterization of the device measuring the displacement of the sonotrode tip and input electrical impedance around excitation frequency with different amounts of the applied electrical power The second method is based on measuring the spatial pressure magnitude distribution of an ultrasound surgical device produced in an anechoic tank. The acoustic reciprocity principle is used to determinate the derived acoustic power of equivalent ultrasound sources at frequency components present in the spectrum of radiated ultrasonic waves. The third method is based on measuring the total absorbed acoustic power in the restricted volume of water using the calorimetric method. In the electromechanical characterization, calculated electroacoustic efficiency factor from equivalent electrical circuits is between 40-60%, the same as one obtained measuring the derived acoustic power in an anechoic tank when there is no cavitation. When cavitation activity is present in the front of the sonotrode tip the bubble cloud has a significant influence on the derived acoustic power and decreases electroacoustic efficiency. The measured output acoustic power using calorimetric method is greater then derived acoustic power, due to a large amount of heat energy released in the cavitation process.     

Phytochemical Composition and Antioxidant Activities of the Essential Oil and Extracts of Satureja subspicata Vis. Growing in Bosnia and Herzegovina

The phytochemical composition and the antioxidant activities of the essential oil, as well as methanol and hot water extracts of endemic Satureja subspicata Vis . growing in Bosnia and Herzegovina (BiH), were described. β‐Caryophyllene, cis‐β‐ocimene, and α‐pinene, identified by GC/MS and GC‐FID, were the dominant oil components. The major compound of both of extracts, identified by HPLC‐DAD, was rosmarinic acid. The analyzed essential oil showed moderate antioxidant activity. In this first report on the extracts of S. subspicata growing in BiH, the obtained results showed a high content of rosmarinic acid, as well as considerable amount of total phenols and flavonoids. Compared to the hot water extract, the methanol extract exhibits higher antioxidant potential, measured by DPPH and FRAP assay (IC₅₀ = 0.45 g/l and 1879.43 equiv. Fe²⁺ μm ), while the hot water extract showed higher potential in inhibition of linoleic acid oxidation (51.7% and 61.5% for 1 and 10 g/l). A good antioxidant potential of the tested extracts indicates their potential use as antioxidants, particularly for lipid protection, and partly explains the justification of the use of this plant in traditional medicine of BiH.