High level synthesis of levan by a novel Halomonas species growing on defined media

A novel halophilic isolate from soil samples taken from Çamalt? Saltern area in Turkey, Halomonas sp. AAD6 (JCM 15723) strain, was found to produce high levels of exopolysaccharides (EPS), in the presence of sucrose in defined media, by flasks and bioreactor condition yielded 1.073 g L^?1 and 1.844 g L^?1, respectively. Sugar analysis, methylation studies and NMR analysis of EPS indicated the repeating unit of this polysaccharide was composed of β-(2,6)-d-fructofuranosyl residues. Hence with this work, Halomonas sp. has been described as a levan producer microorganism for the first time. Biocompatibility studies showed this EPS did not affect cellular viability and proliferation of osteoblasts and murine macrophages. The protective effect of the polymer against the toxic activity of avarol implied its additional use as an anti-cytotoxic agent. Halomonas sp. AAD6 could represent an alternative cheap source of levan polymer when grown on defined media hypothesizing its larger employment in industrial application being non pathogenic microorganism.     

Antiviral efficacy of Limonium densiflorum against HSV-1 and influenza viruses

Viral infections remain a major threat to humans and animals and there is a crucial need for new antiviral agents especially with the development of resistant viruses. Several Limonium genus members (Plumbaginacea) have been widely used in traditional medicine for the treatment of infections. In this study, we investigated the antiviral activities of different fractions after successive extraction (hexane, dichloromethane, ethanol and methanol) of the halophyte Limonium densiflorum against H1N1 influenza and HSV-1 herpes viruses. In addition, TLC phytochemicals of the shoot extracts were analyzed. All extracts were tested for their cytotoxicity using a fluorometric resazurin assay. The antiviral activity of extracts was tested using four modes of action: virucidal test, pretreatment of cells with samples before infection, attachment assay and plaque reduction test. A good antiviral activity was found with ethanol and methanol extracts. They were most potent in HSV-1 inhibition than H1N1 influenza virus. The most potent inhibition was observed with ethanol extract, and it exhibited high levels of virucidal activity against HSV-1 (IC₅₀ = 6 μg/mL). It inhibits the replication of the virus by 75% when added after penetration of the virus, and by 100% when added during the viral attachment. It protects MDCK cells against influenza virus by abolishing virus to entry into the host cell (IC₅₀ = 55 μg/mL). After attachment of influenza virus, the ethanol extract displayed an appreciable inhibition of virus replication (IC₅₀ = 193 μg/mL). Methanol extract showed a moderate antiviral capacity against both viruses. While dichloromethane has excellent antiherpes potential, results were inappropriate because it was toxic to Vero cells, hexane extract has no effect. TLC analysis of these extracts showed that flavonoids and saponins were the major classes of natural products found in the shoot extracts that may be responsible for these antiviral activities.     

Fractionation,characterization and antioxidant activity of exopolysaccharides from fermentation broth of a Cordyceps sinensis fungus

Exopolysaccharide (EPS) fractions P_1/5, P_2/5, P₁, P₂ and P₅ were isolated from the fermentation medium of a medicinal fungus Cordyceps sinensis by gradient precipitation with ethanol at 1/5, 2/5, 1, 2, and 5 volume ratios to the liquid medium. P_1/5 and P_2/5 were mainly composed of polysaccharides with negligible protein content and a large molecular or particle size (intrinsic viscosity [η] 2025 mL/g; hydrodynamic radius R_h 905 nm). The fraction attained at a higher ethanol volume ratio had a much higher protein and lower carbohydrate content, and a smaller molecular size. In particular, P₅ was composed of mainly protein with an average molecular weight 16 kDa, [η] 4.3 mL/g and R_h 23.5 nm. The antioxidant activities of EPS fractions showed a significant dependence on the protein content, being negligible of P_1/5 and P_2/5, low to moderate of P₁ and P₂, and very strong of P₅. Gradient ethanol precipitation was proven a simple and workable method for initial fractionation of polysaccharides, proteins and their complexes with different molecular sizes and for further identification of the bioactive components.     

Enhanced exopolysaccharide production and biological activity of Lactobacillus rhamnosus ZY with calcium and hydrogen peroxide

Exopolysaccharides (EPS) are important food and drug additives with beneficial antioxidant, anticancer, and immune-related effects on human health. However, the EPS is limited by low yields and the need for complex culture conditions in fermentation. Here, we report that hydrogen peroxide and calcium stimulated probiotic activity and production of crude exopolysaccharide (c-EPS) by Lactobacillus rhamnosus ZY. Accordingly, supplementation with 3 mM H₂O₂ allowed c-EPS biosynthesis to reach 567 mg/L after 24 h. Addition of both CaCl₂ and H₂O₂ resulted in a c-EPS yield of 2498 mg/L after 12 h, over 9-fold higher than that of an anaerobic culture. We observed that exposure to calcium and hydrogen peroxide made the cells more hydrophobic and led to the over-expression of GroEL, NADH peroxidase, and glyceraldehyde 3-phosphate dehydrogenase, thus increasing energy storage and EPS production. Chromatographic analysis revealed c-EPS was composed mainly of mannose (5.1%), galactose (15.3%), glucose (20–30%), and rhamnose (50–60%). Preliminary in vitro tests revealed that H₂O₂ and CaCl₂ enhanced the 2,2-diphenyl-1-picrylhydrazyl and hydroxyl radical scavenging capacities, resulting in a notable protective effect against oxidative damage in NIH/3T3 cells. Our study provides a simple and cost-effective approach for achieving high yields of good quality EPS using Lactobacillus rhamnosus.     

Aflatoxin contamination of wheat flour and the risk of esophageal cancer in a high risk area in Iran

Background: Golestan province in northeastern Iran has been known as a high-risk area for esophageal cancer (EC). This study was conducted to assess aflatoxin (AF) contamination of wheat flour (WF) samples in high and low EC-risk areas of Golestan province. Methods: Four WF samples were collected randomly from each of 25 active silos throughout the province in 2009. The levels of AFs were measured using the High-performance liquid chromatography method. Using the data of EC rates obtained from Golestan population-based cancer registry, the province was divided into high and low risk areas for EC. Student t-test and multivariate regression analysis were used to compare the levels of aflatoxins as well as the condition of silos between the two areas. Results: One hundred WF samples were collected. The mean levels of total aflatoxin and aflatoxin B1 was 1.99 and 0.53 ng g^?1, respectively. The levels of total AF (p = 0.03), AFG2 (p = 0.02) and AFB1 (p = 0.003) were significantly higher in samples obtained from high risk area. Multivariate regression analysis showed that humidity of silo was the most important source of difference between silos of the two areas (p = 0.04). Conclusion: We found a positive relationship between AF level of WF samples and the risk of EC. So, AF contamination may be a possible risk factor for EC in our region. We also found that humidity of silos was the most important determinant of AF contamination of WF. Intensive control of silos conditions including humidity and temperature are needed especially in high EC-risk areas.     

Production of a new exopolysaccharide (EPS) by Pseudomonas oleovorans NRRL B-14682 grown on glycerol

Pure glycerol and glycerol-rich product (GRP) obtained from the biodiesel industries were used as carbon source for the production of a new extracellular polysaccharide (EPS) by Pseudomonas oleovorans NRRL B-14682. The influence of temperature (20–40 °C) and pH (6.0–8.0) was studied. A temperature of 30 °C and pH control at 6.8 gave the maximum cell growth and EPS production. The culture attained a maximum cell dry weight (CDW) of 9.55 g l⁻¹ and an EPS concentration of 11.82 g l⁻¹ when cultivated with pure glycerol. GRP was a suitable carbon source, as shown by the slightly higher EPS concentration (12.18 g l⁻¹). The EPS productivity obtained with GRP (3.85 g l⁻¹ d⁻¹) was almost twice that obtained with pure glycerol (2.00 g l⁻¹ d⁻¹). Also, the yield on glycerol was higher for the cultivation with GRP (0.36 g g⁻¹) than for pure glycerol (0.28 g g⁻¹). The EPS was a high molecular weight heteropolysaccharide, composed by neutral sugars (37–80 wt% galactose, 2–30 wt% glucose, 0.5–25 wt% mannose and 0.5–20 wt% rhamnose) and containing acyl group substituents (pyruvil, acetyl and succinyl were identified). The EPS forms highly viscous aqueous dispersions with many potential commercial applications.     

Production,characterization and antioxidant activities in vitro of exopolysaccharides from endophytic bacterium Paenibacillus polymyxa EJS-3

The production, characterization and antioxidant activities in vitro of exopolysaccharides (EPS) from endophytic bacterium Paenibacillus polymyxa EJS-3 were investigated. For EPS production, the preferable culture conditions were 24 °C and pH 8 for 60 h with sucrose and yeast extract as the carbon and nitrogen sources, respectively. Notably, sucrose concentration was the prominent factor, and the maximum yield of EPS (22.82 g/L) was obtained at a sucrose concentration of 160 g/L. The crude EPS was purified by chromatography of DEAE-52 and Sephadex G-100, affording EPS-1 and EPS-2 with molecular weights of 1.22 × 10⁶ and 8.69 × 10⁵ Da, respectively. They were composed of mannose, fructose and glucose in a molar ratio of 2.59:29.83:1 and 4.23:36.59:1, respectively. In addition, both crude and purified EPS showed strong scavenging activities on superoxide and hydroxyl radicals, and their antioxidant activities decreased in the order of crude EPS > EPS-2 > EPS-1.     

Emulsifying properties of a marine bacterial exopolysaccharide

Exopolysaccharide produced by a marine Enterobacter cloaceae (designated as EPS 71a) emulsified hexane, benzene, xylene, kerosene, paraffin oil, cottonseed oil, coconut oil, jojoba oil, castor oil, groundnut oil and sunflower oil. However, it could form stable emulsions with groundnut oil and hexane at optimal concentration of 1 mg ml⁻¹. Further increase in concentration of EPS 71a did not show noteworthy increase in emulsification indices. Emulsions with groundnut oil and hexane were stable up to 10 days between pH 2 and 10 and in the presence of sodium chloride in the range of 5–50 mg ml⁻¹ at 35–37 °C. EPS 71a formed stable emulsion with xylene as compared to commercial gums such as arabic, tragacanth, karaya and xanthan.     

Characterization and antioxidant activity for exopolysaccharide from submerged culture of Boletus aereus

Three polysaccharide fractions (designated as Fr-I, Fr-II and Fr-III) were successfully purified from the crude exopolysaccharide (EPS) produced from submerged culture of Boletus aereus by gel filtration chromatography on Sepharose CL-6B. The size exclusion chromatography/multi-angle laser light scattering (SEC/MALLS) system showed that the average molecular weights (Mws) of these three fractions were 1.365 × 10⁶, 1.048 × 10⁵ and 2.471 × 10⁴ g/mol, respectively. The SEC/MALLS also revealed that the molecular conformation of the Fr-I was a random coil, with Fr-II being a rigid rod in aqueous solution. Moreover, monosaccharide composition analysis indicated that Fr-I was mainly composed of glucose, while both of Fr-II and Fr-III were mainly composed of mannose and glucose. Then, FT-IR spectral analysis of the purified EPS revealed prominent characteristic groups. Furthermore, thermo gravimetric analysis (TGA) indicated the degradation temperature of the Fr-I (170 °C) was higher than those of Fr-II (156 °C) and Fr-III (155 °C). Finally, on the basis of the antioxidant activity test in vitro, Fr-I exhibited the highest antioxidant ability among these samples, which might be attributed to the monosaccharide composition and molecular weight in the EPS fraction.     

Noninvasive genetic assessment of brown bear population structure in Bulgarian mountain regions

The Balkans are one of the last large refugia for brown bear (Ursus arctos) populations in Europe, and Bulgaria, in particular, contains relatively large areas of suitable brown bear habitat and a potential population of more than 600 individuals. Despite this, the majority of brown bear research remains focused on bear populations in Central and Western Europe. We provide the first assessment of genetic population structure of brown bears in Bulgaria by analysing tissue samples (n = 16) as well as samples collected with noninvasive genetic methods, including hair and faecal samples (n = 189 and n = 163, respectively). Sequence analysis of a 248 base pair fragment of the mitochondrial control region showed that two highly divergent mitochondrial European brown bear lineages form a contact zone in central Bulgaria. Furthermore, the analysis of 13 polymorphic microsatellite markers identified 136 individuals and found substantial genetic variability (H_e = 0.74; N_A = 8.9). The combination of both genetic markers revealed the presence of weak genetic substructure in the study area with considerable degrees of genetic admixture and the likely presence of migration corridors between the two subpopulation in the Rhodope Mountains and Stara Planina as evidenced from the genetic detection of two male long-distance dispersers. A detailed assessment from densely collected samples in the Rhodope Mountains resulted in a population size estimate of 315 (95% CI = 206–334) individuals, indicating that not all available habitat is presently occupied by bears in this region. Efficient management plans should focus on preserving connectivity of suitable habitats in order to maintain gene flow between the two Bulgarian brown bear subpopulations.     

An in situ study of bioenergetic properties of human colorectal cancer: The regulation of mitochondrial respiration and distribution of flux control among the components of ATP synthasome

The aim of this study is to characterize the function of mitochondria and main energy fluxes in human colorectal cancer (HCC) cells. We have performed quantitative analysis of cellular respiration in post-operative tissue samples collected from 42 cancer patients. Permeabilized tumor tissue in combination with high resolution respirometry was used.Our results indicate that HCC is not a pure glycolytic tumor and the oxidative phosphorylation (OXPHOS) system may be the main provider of ATP in these tumor cells. The apparent Michaelis–Menten constant (K_m) for ADP and maximal respiratory rate (V_m) values were calculated for the characterization of the affinity of mitochondria for exogenous ADP: normal colon tissue displayed low affinity (K_m = 260 ± 55 μM) whereas the affinity of tumor mitochondria was significantly higher (K_m = 126 ± 17 μM). But concurrently the V_m value of the tumor samples was 60–80% higher than that in control tissue. The reason for this change is related to the increased number of mitochondria. Our data suggest that in both HCC and normal intestinal cells tubulin β-II isoform probably does not play a role in the regulation of permeability of the MOM for adenine nucleotides.The mitochondrial creatine kinase energy transfer system is not functional in HCC and our experiments showed that adenylate kinase reactions could play an important role in the maintenance of energy homeostasis in colorectal carcinomas instead of creatine kinase.Immunofluorescent studies showed that hexokinase 2 (HK-2) was associated with mitochondria in HCC cells, but during carcinogenesis the total activity of HK did not change. Furthermore, only minor alterations in the expression of HK-1 and HK-2 isoforms have been observed.Metabolic Control analysis showed that the distribution of the control over electron transport chain and ATP synthasome complexes seemed to be similar in both tumor and control tissues. High flux control coefficients point to the possibility that the mitochondrial respiratory chain is reorganized in some way or assembled into large supercomplexes in both tissues.     

Relationship between the resistance to bile salts and low pH with exopolysaccharide (EPS) production of Bifidobacterium spp. isolated from infants feces and breast milk

The purpose of this study was to investigate a possible relation between resistance to bile salts and low pH with exopolysaccharide (EPS) producing of Bifidobacterium spp. In this study, a total of 31 Bifidobacterium spp. were isolated from breast fed infants feces and breast milk samples. As a result of the identification tests, isolates were identified as Bifidobacterium breve (15 strains), B. bifidum (11 strains), B. pseudocatenulatum (3 strains) and B. longum (2 strains). Bifidobacterium spp. were determined exopolysaccharide (EPS) production. EPS productions observed at chance rations (38.00–97.64 mg/l) among of Bifidobacterium spp. Furthermore, Bifidobacterium spp. were determined resistance to bile salts and low pH. Positive correlations between production of exopolysaccharide and resistance to bile salts (p < 0.01) or low pH (p < 0.01) were found Bifidobacterium spp. This investigation showed that high EPS production of Bifidobacteria may be important in the selection of probiotic strains for resistance to bile salts and low pH.     

Design,synthesis, antiviral and cytostatic activity of ω-(1H-1,2,3-triazol-1-yl)(polyhydroxy)alkylphosphonates as acyclic nucleotide analogues

The efficient synthesis of a new series of polyhydroxylated dibenzyl ω-(1H-1,2,3-triazol-1-yl)alkylphosphonates as acyclic nucleotide analogues is described starting from dibenzyl ω-azido(polyhydroxy)alkylphosphonates and selected alkynes under microwave irradiation. Selected O,O-dibenzylphosphonate acyclonucleotides were transformed into the respective phosphonic acids. All compounds were evaluated in vitro for activity against a broad variety of DNA and RNA viruses and for cytostatic activity against murine leukemia L1210, human T-lymphocyte CEM and human cervix carcinoma HeLa cells. Compound (1S,2S)-16b exhibited antiviral activity against Influenza A H3N2 subtype (EC₅₀ = 20 μM—visual CPE score; EC₅₀ = 18 μM—MTS method; MCC >100 μM, CC₅₀ >100 μM) in Madin Darby canine kidney cell cultures (MDCK), and (1S,2S)-16k was active against vesicular stomatitis virus and respiratory syncytial virus in HeLa cells (EC₅₀ = 9 and 12 μM, respectively). Moreover, compound (1R,2S)-16l showed activity against both herpes simplex viruses (HSV-1, HSV-2) in HEL cell cultures (EC₅₀ = 2.9 and 4 μM, respectively) and feline herpes virus in CRFK cells (EC₅₀ = 4 μM) but at the same time it exhibited cytotoxicity toward uninfected cell (MCC ⩾ 4 μM). Several other compounds have been found to inhibit proliferation of L1210, CEM as well as HeLa cells with IC₅₀ in the 4–50 μM range. Among them compounds (1S,2S)- and (1R,2S)-16l were the most active (IC₅₀ in the 4–7 μM range).     

Optimization of the aeration and agitation speed of Aeribacillus palidus 418 exopolysaccharide production and the emulsifying properties of the product

The specific properties of exopolysaccharides (EPS) from thermophilic microorganisms have attracted interest in their optimized production. In this study, the ability of Aeribacillus pallidus 418 to grow and produce polysaccharide in a 5-l stirred tank bioreactor was investigated. Agitation rates of 100, 200, 600, 900, and 1100 revolutions per minute (rpm), at an air flow rate of 0.5 gas volumes per unit medium volume per minute (vvm), and aeration rates of 0.25, 0.5, 1.0, and 1.5 vvm, at an agitation rate of 900 rpm, were examined. A maximum EPS yield of 170 μg/ml has been registered in a single impeller bioreactor equipped with an original Narcissus impeller at agitation speed of 900 rpm, with an aeration rate of 0.5 vvm. The bioprocess oxygen uptake rate (OUR) and oxygen mass transfer coefficient (K_La) were evaluated. The emulsifying properties of the specific EPS produced by A. pallidus 418 were determined. Stable oil-in-water emulsions, a low level of separated water phase and high dispersion stability were found, which together demonstrate the prospects for the industrial exploration of EPS production. Enhanced synergism between the A. pallidus 418 synthesized EPS and various commercially used hydrocolloids was observed; superior synergy was achieved in combination with xanthan gum.     

Sex-specific light acclimation of Chara canescens (Charophyta)

Bisexual populations of the charophyte Chara canescens (Desv. et Loisel. in Loisel., 1810) containing male and female individuals are rarely found. Two experiments were carried out to study whether male and female algae from the same site exhibit different physiological capacities, especially with respect to light acclimation.Algae from two different shore levels and from laboratory cultures acclimated to six irradiance conditions (35–500 μmol photons m⁻² s⁻¹) were compared. Field measurements showed that both female and male algae of C. canescens are able to acclimate to daily changes in solar irradiance. The quantum yield of Photosystem II (PSII) decreased with increasing irradiance in the morning and increased with decreasing irradiance in the afternoon. Growth experiments showed increasing growth rates from 35 μmol photons m⁻² s⁻¹ (∼7 mg FW) up to 500 μmol photons m⁻² s⁻¹ (∼27 mg FW) in female and male C. canescens. The irradiance saturation point for photosynthesis (E_k) was about 140 μmol m⁻² s⁻¹ for both sexes within the whole range of acclimation irradiances. The maximum photosynthesis rate at saturating irradiances (P_max) of male algae was highest at E_k, whereas P_max of female algae was highest at 500 μmol photons m⁻² s⁻¹. The photosynthetic efficiency in the light-limited range (α) increased in female C. canescens and decreased in male C. canescens. The ratio of the non-photochemical quenching parameter (NPQ) to the relative electron transport rates rETR_(MT) increased in both sexes with irradiance, but showed a steeper increase in male than in female algae. Pigment analysis showed similar acclimation pattern for male and female C. canescens. Chl a/Chl b ratios of both sexes were constant over the whole range of E_g, whereas Chl a/carotenoid ratios in male and female C. canescens decreased from 70 μmol photons m⁻² s⁻¹ upwards. Pigment analysis pointed out that the carotenes α-, β- and γ-carotene were more prominent in male than in female algae.Our results indicate that female C. canescens are more efficient in light acclimation than male algae from the same site. Nevertheless, further investigations of bisexual C. canescens populations resolving CO₂-uptake mechanisms and/or genetic differences are needed.     

Toxicokinetics and toxicological effects of single oral dose of fumonisin B1 containing Fusarium verticillioides culture material in weaned piglets

Toxicokinetics and the toxicological effects of culture material containing fumonisin B₁ (FB₁) were studied in male weaned piglets by clinical, pathological, biochemical and sphingolipid analyses. The animals received a single oral dose of 5 mg FB₁/kg of body weight, obtained from Fusarium verticillioides culture material. FB₁ was detected by HPLC in plasma collected at 1-h intervals up to 6 h and at 12-h intervals up to 96 h. FB₁ eliminated in feces and urine was quantified over a 96-h period and in liver samples collected 96 h post-intoxication. Blood samples were obtained at the beginning and end of the experiment to determine serum enzyme activity, total bilirubin, cholesterol, sphinganine (Sa), sphingosine (So) and the Sa/So ratio. FB₁ was detected in plasma between 30 min and 36 h after administration. The highest concentration of FB₁ was observed after 2 h, with a mean concentration of 282 μg/ml. Only 0.93% of the total FB₁ was detected in urine between 75 min and 41 h after administration, the highest mean concentration (561 μg/ml) was observed during the interval after 8 at 24 h. Approximately 76.5% of FB₁ was detected in feces eliminated between 8 and 84 h after administration, with the highest levels observed between 8 and 24 h. Considering the biochemical parameters, a significant increase only occurred in cholesterol, alkaline phosphatase and aspartate aminotransferase activities. In plasma and urine, the highest Sa and Sa/So ratios were obtained at 12 and 48 h, respectively.     

Prevalence,distribution and intraspecific variation of Heterodera schachtii populations from semiarid environment

A two-year survey study was carried-out to identify and determine the distribution, community characteristics and intraspecific variation of the sugar beet cyst nematode, Heterodera schachtii populations collected from the six major sugar beet-producing governorates in Syria, west Asia. A total of 178 composite rhizosphere soil samples were collected from the six governorates during 2009 and 2010 growing seasons as follows: Aleppo (26), Ar Raqqah (17), Dayr az Zawr (33), Idlib (34), Hamah (41) and Homs (27 samples). The nematode was originally identified on the basis of morphometrics and morphological features of the cyst vulvar cones and second-stage juveniles (J₂). The communities of H. schachtii were then analyzed using the criteria of frequency of occurrence (FO%), mean population density (PD) and prominence value (PV). Results showed that H. schachtii was the most frequent and prominent in Homs (FO% = 70.37% and PV = 23472.37) and the least frequent and prominent in Ar Raqqah (FO% = 5.88% and PV = 28.86). The highest nematode population density was also recorded in Homs in 2009 (50,545 J₂ + eggs/100 g soil) while the least was recorded in Ar Raqqah in 2010 (119 J₂ + eggs/100 g soil). The morphometric data of cyst vulvar cone, second-stage juveniles (J₂) and eggs were also subjected to a multivariate principal component analysis to analyze the relationships between the studied populations, and to identify the variables that show the highest multiple correlations with these populations. Results showed that at the high order of hierarchical cluster analysis, four populations (Idlib, Hamah, Homs and Aleppo) formed one main cluster, while Dair az Zawr and Ar Raqqah failed to form clusters and separated individually. However, at low distance value, the main cluster further subdivided to separate the Aleppo population from Idlib, Hamah and Homs populations. The morphometric distance values ranged from 0.07 to 0.61. Hamah and Homs were the closest populations while Dair az Zawr showed the most diverse population. This study also revealed the value of J₂ body length, egg length and width, fenestral length and width and the number of bullae inside the cyst vulvar cone in determining the intraspecific variations among the Syrian populations of H. schachtii.     

Structural changes that occur upon photolysis of the Fe(II)a3–CO complex in the cytochrome ba3-oxidase of Thermus thermophilus: A combined X-ray crystallographic and infrared spectral study demonstrates CO binding to CuB

The purpose of the work was to provide a crystallographic demonstration of the venerable idea that CO photolyzed from ferrous heme-a₃ moves to the nearby cuprous ion in the cytochrome c oxidases. Crystal structures of CO-bound cytochrome ba₃-oxidase from Thermus thermophilus, determined at ~ 2.8–3.2 Å resolution, reveal a Fe–C distance of ~ 2.0 Å, a Cu–O distance of 2.4 Å and a Fe–C–O angle of ~ 126°. Upon photodissociation at 100 K, X-ray structures indicate loss of Fe_a3–CO and appearance of Cu_B–CO having a Cu–C distance of ~ 1.9 Å and an O–Fe distance of ~ 2.3 Å. Absolute FTIR spectra recorded from single crystals of reduced ba₃–CO that had not been exposed to X-ray radiation, showed several peaks around 1975 cm^? 1; after photolysis at 100 K, the absolute FTIR spectra also showed a significant peak at 2050 cm^? 1. Analysis of the ‘light’ minus ‘dark’ difference spectra showed four very sharp CO stretching bands at 1970 cm^? 1, 1977 cm^? 1, 1981 cm^? 1, and 1985 cm^? 1, previously assigned to the Fe_a3–CO complex, and a significantly broader CO stretching band centered at ~ 2050 cm^? 1, previously assigned to the CO stretching frequency of Cu_B bound CO. As expected for light propagating along the tetragonal axis of the P4₃2₁2 space group, the single crystal spectra exhibit negligible dichroism. Absolute FTIR spectrometry of a CO-laden ba₃ crystal, exposed to an amount of X-ray radiation required to obtain structural data sets before FTIR characterization, showed a significant signal due to photogenerated CO₂ at 2337 cm^? 1 and one from traces of CO at 2133 cm^? 1; while bands associated with CO bound to either Fe_a3 or to Cu_B in “light” minus “dark” FTIR difference spectra shifted and broadened in response to X-ray exposure. In spite of considerable radiation damage to the crystals, both X-ray analysis at 2.8 and 3.2 Å and FTIR spectra support the long-held position that photolysis of Fe_a3–CO in cytochrome c oxidases leads to significant trapping of the CO on the Cu_B atom; Fe_a3 and Cu_B ligation, at the resolutions reported here, are otherwise unaltered. This article is part of a Special Issue entitled: Respiratory Oxidases.     

Picrasin K,a new quassinoid from Quassia amara L. (Simaroubaceae)

A new quassinoid Picrasin K 1 was isolated from a decoction made of Quassia amara leaves, traditionally used in French Guyana to treat malaria. The structure and relative stereochemistry of 1 was determined through extensive NMR analysis. Picrasin K showed a low activity against Plasmodium falciparum in vitro (IC₅₀ = 8 μM), and a similar low activity on human cancerous cells line (IC₅₀ = 7 μM on MCF-7 cells line).     

Chemical composition,angiotensin I-converting enzyme (ACE) inhibitory,antioxidant and antimicrobial activities of the essential oil from south Tunisian Ajuga pseudoiva Rob. Lamiaceae

The essential oil of Ajuga pseudoiva, collected from Tunisia, was analyzed using gas chromatography–mass spectroscopy. Thirty-two compounds accounting for 95.76% of the total oil were identified. Sesquiterpenes were found to be the most abundant components of A. pseudoiva oil. And they were mainly represented by viridiflorol (30.17%), germacrene B (9.26%) (α, β and γ)-eudesmol (8.11%) and aromadendrene (7.45%). The essential oil of A. pseudoiva showed radical scavengers activity (IC₅₀ = 0.72 mg/mL) and displayed lipid peroxidation inhibitory activity (IC₅₀ = 0.6 mg/mL). A. pseudoiva essential oil was also found to exhibit a dose-dependent ACE inhibitory activity with an IC₅₀ value of 65.5 μg/mL. Moreover, the antimicrobial activity of the essential oil was tested against 17 species of microorganisms, and the results obtained showed significant antibacterial activity against the Gram-positive and Gram-negative bacteria, with inhibition zones and minimal inhibitory concentration values of 14–32 mm and 84–137 μg/mL and 6–21 mm and 105–336 μg/mL, respectively. Higher activity was also found against several fungal strains.