Effects of oxidation on the hydrolysis by cholesterol esterase of sitosteryl esters as compared to a cholesteryl ester

Phytosteryl esters (PE) are used as ingredients in functional food to decrease plasma concentration of low density lipoprotein-cholesterol (LDL-C). Effective impairment of cholesterol absorption by PE suggests that these esters are hydrolyzed by the pancreatic cholesterol esterase (CEase, EC 3.1.1.13) and the liberated sterol may interfere with cholesterol reducing its intestinal absorption. PE-enriched foods are marketed for cooking purposes, and temperature is one of the most important factors leading to the formation of oxidation products. Very little is known about the outcome of PE oxides during the digestive process. A new analytical method based on mass spectrometric detection directly after enzymatic reaction was developed to determine in vitro the activity of CEase on PE and their oxides present in functional food. Using this method, we identified a new inhibitor of CEase: sitosteryl 9,10-dihydroxystearate, which behaves as a non-competitive inhibitor of the hydrolysis of cholesteryl oleate and sitosteryl oleate.     

New cholesterol esterase inhibitors based on rhodanine and thiazolidinedione scaffolds

We present a new class of inhibitors of pancreatic cholesterol esterase (CEase) based on ‘priviledged’ 5-benzylidenerhodanine and 5-benzylidene-2,4-thiazolidinedione structural scaffolds. The lead structures (5-benzylidenerhodanine 4a and 5-benzylidene-2,4-thiazolidinedione 4b) were identified in an in-house screening and these inhibited CEase with some selectivity over another serine hydrolase, acetylcholinesterase (AChE) (4a, CEase IC₅₀ = 1.76 μM vs AChE IC₅₀ = 5.14 μM and 4b, CEase IC₅₀ = 5.89 μM vs AChE IC₅₀ >100 μM). A small library of analogs (5a–10a) containing a core amino acid in place of the glycerol group of the lead structures, was prepared to explore other potential binding interaction with CEase. These analogs inhibited CEase with IC₅₀ values ranging from 1.44 to 85 μM, with the majority exhibiting some selectivity for CEase versus AChE. The most potent compound of the library (10a) had 17-fold selectivity over AChE. We also report molecular docking (with CEase) and detailed kinetic analysis on the amino acid analogs to further understand the associated structure–activity relationships.     

Cholesterol esterase action on human high density lipoproteins and inhibition studies: detection by MALDI-TOF MS

The modification of lipoproteins by lipolytic enzymes such as cholesterol esterase (CEase) is assumed to play an important role in the pathogenesis of atherosclerosis. However, details of the activation and inhibition of CEase are still unknown. In this study, matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to investigate the extracts of human lipoproteins after treatment with CEase and to monitor the effects of the inhibitor 2-(diethylamino)-6,7-dihydro-4H,5H-cyclopenta[4,5] thieno[2,3-d][1,3]oxazin-4-one (DOT-3). This approach has the advantage that all lipid classes can be independently detected; therefore, conclusions on the mechanism of the applied enzyme are possible. Besides the expected decrease of cholesteryl esters (CEs) in HDL, a significantly enhanced content of lysophosphatidylcholine (LPC) was also detected, confirming the broad substrate specificity of CEase. It was also demonstrated that DOT-3 significantly inhibited the CEase-catalyzed cleavage of CEs in HDL. Phospholipid (PL) vesicles prepared from phosphatidylcholine (PC) or PC and cholesteryl linoleate were treated with CEase, and the changes in lipid composition were investigated. From the analysis of the generated LPC species in HDL and in the isolated lipid mixtures, it is evident that CEase catalyzes the cleavage of the fatty acid residues in both the sn-1 and sn-2 positions of the PLs. These effects are obvious in the absence as well as in the presence of detergents.     

Isocoumarin-based inhibitors of pancreatic cholesterol esterase

Pancreatic cholesterol esterase (CEase), which is secreted from the exocrine pancreas, is a serine hydrolase that aids in the bile salt-dependent hydrolysis of dietary cholesteryl esters and contributes to the hydrolysis of triglycerides and phospholipids. Additional roles for CEase in intestinal micelle formation and in transport of free cholesterol to the enterocyte have been suggested. There also are studies that point to a pathological role(s) for CEase in the circulation where CEase accumulates in atherosclerotic lesions and triggers proliferation of smooth muscle cells. Thus, there is interest in CEase as a potential drug target. 4-Chloro-3-alkoxyisocoumarins are a class of haloenol lactones that inhibit serine hydrolases and serine proteases and have the potential to be suicide inhibitors. In the present study, we have developed 3-alkoxychloroisocoumarins that are potent inhibitors of CEase. These inhibitors were designed to have a saturated cycloalkane ring incorporated into a 3-alkoxy substituent. The size of the ring as well as the length of the tether holding the ring was found to be important contributors to binding to CEase. 4-Chloro-3-(4-cyclohexylbutoxy)isocoumarin and 4-chloro-3-(3-cyclopentylpropoxy)isocoumarin were demonstrated to be potent reversible inhibitors of CEase, with dissociation constants of 11 nM and 19 nM, respectively. The kinetic results are consistent with predictions from molecular modeling.     

Benzoflavones as cholesterol esterase inhibitors: Synthesis,biological evaluation and docking studies

A library of forty 7,8-benzoflavone derivatives was synthesized and evaluated for their inhibitory potential against cholesterol esterase (CEase). Among all the synthesized compounds seven benzoflavone derivatives (A-7, A-8, A-10, A-11, A-12, A-13, A-15) exhibited significant inhibition against CEase in in vitro enzymatic assay. Compound A-12 showed the most promising activity with IC₅₀ value of 0.78 nM against cholesterol esterase. Enzyme kinetic studies carried out for A-12, revealed its mixed-type inhibition approach. Molecular protein–ligand docking studies were also performed to figure out the key binding interactions of A-12 with the amino acid residues of the enzyme’s active site. The A-12 fits well at the catalytic site and is stabilized by hydrophobic interactions. It completely blocks the catalytic assembly of CEase and prevents it to participate in ester hydrolysis mechanism. The favorable binding conformation of A-12 suggests its prevailing role as CEase inhibitor.     

Kinetics and mechanisms of cholesterol esterase inhibition by cardiovascular drugs in vitro

Cardiovascular drugs such as lovastatin, simvastatin, amlodipine besylate, nifedipine, and hydralazine hydrochloride inhibit cholesterol esterase (CEase) in vitro. In the present paper, an attempt was made to determine kinetically the reaction mechanism for CEase inhibition by these drugs. The inhibition constant, Ki, for the mixed-type inhibition of CEase by these drugs in the presence of triton-X-100 or taurochloate were measured. Moreover, the pKi values were correlated with the molecular weights of these drugs. In conclusion, the fact that these drugs lower cholesterol levels in the plasma low-density lipoprotein may be partially due to the CEase inhibition by these drugs.     

Effects of oxidation on the hydrolysis by cholesterol esterase of sitosteryl esters as compared to a cholesteryl ester

Phytosteryl esters (PE) are used as ingredients in functional food to decrease plasma concentration of low density lipoprotein-cholesterol (LDL-C). Effective impairment of cholesterol absorption by PE suggests that these esters are hydrolyzed by the pancreatic cholesterol esterase (CEase, EC 3.1.1.13) and the liberated sterol may interfere with cholesterol reducing its intestinal absorption. PE-enriched foods are marketed for cooking purposes, and temperature is one of the most important factors leading to the formation of oxidation products. Very little is known about the outcome of PE oxides during the digestive process. A new analytical method based on mass spectrometric detection directly after enzymatic reaction was developed to determine in vitro the activity of CEase on PE and their oxides present in functional food. Using this method, we identified a new inhibitor of CEase: sitosteryl 9,10-dihydroxystearate, which behaves as a non-competitive inhibitor of the hydrolysis of cholesteryl oleate and sitosteryl oleate.     

Changes in community structure and photosynthetic activities of total phytoplankton species during the growth,maintenance, and dissipation phases of a Prorocentrum donghaiense bloom

The potential interactions between the bloom-forming dinoflagellates and other phytoplankton during the algal bloom cycle are interesting, while the causes for the phytoplankton community changes were not fully understood. We hypothesized that phytoplankton community structure and photosynthetic activities of total phytoplankton have their special characteristics in different phases of the algal blooms. To test this hypothesis, a survey covering the process of a Prorocentrum donghaiense bloom in coastal waters between Dongtou and Nanji Islands was carried out from 9 to 20 May 2016, and the changes in the phytoplankton community and photosynthetic activities of total phytoplankton were determined. Surface seawater was sampled for microscopic analysis of phytoplankton composition and pulse amplitude modulated (PAM) chlorophyll fluorescence analysis of photosynthetic activities of the total phytoplankton species. A total of 25, 31, and 19 phytoplankton species were identified in its growth (9–12 May), maintenance (13–18 May) and dissipation phases (19–20 May), respectively. Diatoms were dominant in terms of species number while dinoflagellates were predominant at cell abundance. Dinoflagellates were the major dominant species during three phases of the bloom based on the dominance (Y) value, whereas the dominant species extended to dinoflagellates and diatoms including P. donghaiense, Coscinodiscus argus, Gonyaulax spinifera, Cyclotella sp. and Scrippsiella trochoidea in the dissipation phase. In the maintenance phase, the average cell abundances of the total phytoplankton and P. donghaiense were consistent with the chlorophyll a (Chla) concentration in the seawater; for the diversity indices of total phytoplankton species, Simpson index (C) was the highest while Shannon index (H′) and Pielou evenness index (J′) were the lowest. Furthermore, photosynthetic activities of the total phytoplankton species represented by the effective quantum yield (F_q'/F_m') and the maximum relative electron transport rate (rETR_max) in the maintenance phase were higher than those in the growth and dissipation phases. The results indicated that the characteristics of phytoplankton community structure and photosynthetic activities could be regarded as criteria in predicting the phases of algal blooms.     

Antioxidant status of Lobiger serradifalci and Oxynoe olivacea (Opisthobranchia, Mollusca)

Invasion of the Mediterranean Sea by the two world-wide famous exotic algae species, Caulerpa taxifolia and Caulerpa racemosa, is still a problem and has adverse effects on the Mediterranean sublittoral ecosystem. Biological control studies revealed that the two native Sacoglossans, Oxynoe olivacea and Lobiger serradifalci, may have an effect on the expansion of invasive Caulerpa spp. in the Mediterranean. In the framework of this study, antioxidant enzyme activities, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), lipid peroxidation (LPO) and oxidized glutathione (GSSG) levels, as oxidative stress markers in L. serradifalci and O. olivacea were determined at two different temperature conditions (20 and 27 °C). In both species, SOD, CAT and GSH-Px activities were found to be positively correlated with temperature. The SOD activities in L. serradifalci were higher than those in O. olivacea at both temperatures, whereas the CAT and GSH-Px activities were significantly (p<0.05) higher in O. olivacea, compared to L. serradifalci. As expected, both species showed decreased LPO levels at 27 °C compared to 20 °C. GSSG level at 27 °C in O.olivacea was significantly (p<0.05) higher than that of 20 °C. On the other hand, no statistical (p>0.05) difference in L.serradifalci existed between GSSG levels at two temperatures. But, despite the variations in the antioxidant enzyme activities, there was no significant difference in LPO levels between the species, suggesting that the oxidative consequences of a given environmental condition may vary among different species. Inasmuch as the GSSG levels were in accordance with antioxidant enzyme activities, GSH might have acted as a cofactor of GSH-Px and an individual antioxidant in these sea slugs.     

Comparative xenobiotic metabolism capacities and pesticide sensitivity in adults of Solea solea and Solea senegalensis

The measurement of enzymatic activities involved in xenobiotic biotransformation was carried out in adults of Solea solea and Solea senegalensis. The hepatic enzymes analysed were cytochrome P450 (CYP) related activities using eight fluorometric substrates and carboxylesterases (CbE). The conjugating activities of glutathione S-transferase (GST) and UPD-glucuronosyltransferase (UDPGT) were also assessed. Specific mammalian inhibitors were used as diagnostic tools for related activities of CYP1A (α-naphthoflavone; αNF), CYP2B6 and CYP2C19 (ticlopidine) and CYP3A4 (ketoconazole). The in vitro sensitivity to organophosphorous pesticides (OP) was tested in the S10 homogenate of brain (acetylcholinesterase-AChE) and liver (CbE). Furthermore, the pesticide chlorpyrifos oxon (CLPO) was used to explore the OP sensitivity of CbE of both species in two subcellular fractions (microsomes and cytosol), using two substrates. Overall, only two parameters confirmed species differences: EROD and cytosolic CbE being significantly elevated (p < 0.05) in the common sole, S. solea. A high inhibition of CYP1A related activities using several fluorometric substrates (ER, MR and CEC) after in vitro incubation with αNF confirmed all measure CYP1A1-related activities whereas ketoconazole was more specific for BFCOD (CYP3A4). Pesticide sensitivity was similar for brain AChE but hepatic CbE had a protective role that was species and pesticide dependent.     

Exogenous jasmonic acid modulates the physiology,antioxidant defense and glyoxalase systems in imparting drought stress tolerance in different Brassica species

This study examined the ability of jasmonic acid (JA) to enhance drought tolerance in different Brassica species in terms of physiological parameters, antioxidants defense, and glyoxalase system. Ten-day-old seedlings were exposed to drought (15 % polyethylene glycol, PEG-6000) either alone or in combination with 0.5 mM JA. Drought significantly increased lipoxygenase activity and oxidative stress, levels of malondialdehyde and H₂O₂. Drought reduced seedling biomass, chlorophyll (chl) content, and leaf relative water content (RWC). Drought increased proline, oxidized ascorbate (DHA) and glutathione disulfide (GSSG) levels. Drought affected different species differently: in B. napus, catalase (CAT) and glyoxalase II (Gly II) activities were decreased, while glutathione-S-transferase (GST) and glutathione peroxidase (GPX) activities were increased in drought-stressed compared to unstressed plants; in B. campestris, activities of glutathione reductase (GR), glyoxalase I (Gly I), GST, and GPX were increased, monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), CAT and other enzymes were decreased; in B. juncea, activities of ascorbate peroxidase, GR, GPX, Gly I were increased; Gly II activity was decreased and other enzymes did not change. Spraying drought-stressed seedlings with JA increased GR and Gly I activities in B. napus; increased MDHAR activity in B. campestris; and increased DHAR, GR, GPX, Gly I and Gly II activities in B. juncea. JA improved fresh weight, chl, RWC in all species, dry weight increased only in B. juncea. Brassica juncea had the lowest oxidative stress under drought, indicating its natural drought tolerance capacity. The JA improved drought tolerance of B. juncea to the highest level among studied species.     

Comparative study of the antioxidant defence systems in the erythrocytes of australian marsupials and monotremes

A comparison of the erythrocyte (RBC) antioxidant metabolites and enzymes in nine marsupial and two monotreme species was carried out. Reduced glutathione (GSH) concentrations were comparable with those reported for other marsupial and eutherian species. An important finding was that the erythrocytes of the southern hairy nosed wombat regenerated GSH faster than the erythrocytes from its close relative, the common wombat. The activities of glutathione-S-transferase, NADH-methaemoglobin reductase, Superoxide dismutase, and glutathione peroxidase (GSH-Px), showed similar levels and extents of variation as those observed in other marsupial and eutherian species. Catalase activities in the marsupials were lower than those measured in the two monotreme species and much lower than those reported in eutherian species. A negative correlation, significant at P < 0.05, was observed between GSH-Px and catalase activities in the RBC of the marsupials. Since both these enzymes “detoxify” H₂O₂, there appears to be a reciprocal relationship between the activities of these enzymes in marsupial RBC     

The effect of pCO2 on carbon acquisition and intracellular assimilation in four marine diatoms

The effect of pCO₂ on carbon acquisition and intracellular assimilation was investigated in the three bloom-forming diatom species, Eucampia zodiacus (Ehrenberg), Skeletonema costatum (Greville) Cleve, Thalassionema nitzschioides (Grunow) Mereschkowsky and the non-bloom-forming Thalassiosira pseudonana (Hust.) Hasle and Heimdal. In vivo activities of carbonic anhydrase (CA), photosynthetic O₂ evolution, CO₂ and HCO₃⁻ uptake rates were measured by membrane-inlet mass spectrometry (MIMS) in cells acclimated to pCO₂ levels of 370 and 800 μatm. To investigate whether the cells operate a C₄-like pathway, activities of ribulose-1,5-bisphosphate carboxylase (RubisCO) and phosphoenolpyruvate carboxylase (PEPC) were measured at the mentioned pCO₂ levels and a lower pCO₂ level of 50 μatm. In the bloom-forming species, extracellular CA activities strongly increased with decreasing CO₂ supply while constantly low activities were obtained for T. pseudonana. Half-saturation concentrations (K_1/2) for photosynthetic O₂ evolution decreased with decreasing CO₂ supply in the two bloom-forming species S. costatum and T. nitzschioides, but not in T. pseudonana and E. zodiacus. With the exception of S. costatum, maximum rates (V_max) of photosynthesis remained constant in all investigated diatom species. Independent of the pCO₂ level, PEPC activities were significantly lower than those for RubisCO, averaging generally less than 3%. All examined diatom species operate highly efficient CCMs under ambient and high pCO₂, but differ strongly in the degree of regulation of individual components of the CCM such as C_i uptake kinetics and extracellular CA activities. The present data do not suggest C₄ metabolism in the investigated species.     

Cholesterol ester hydrolysis in isolated rat liver cells. Effect of estradiol, dibutyryl cyclic AMP and free fatty acids

The basal activity of neutral cholesterol esterase (CEase) in isolated hepatocytes from fed rats showed a characteristic variation during incubation with an initial rise at 30 min and subsequent significant decrease at 2 h. Addition of estradiol (10(-4)M) not only prevented this decrease but also caused an early increase in the activity of the enzyme (125% above basal value) after 60 min of incubation. The simultaneous addition of dibutyryl cyclic AMP (db-cAMP) (10(-5)M) blocked the induction, whereas the cyclic nucleotide alone inhibited significantly the esterase activity. When hepatocyte suspensions were incubated in the presence of different fatty acids (oleate, myristate and laurate, initial concentration 1 mM) for a period of 1 h, a reduction in the CEase activity was observed. The present results suggest that the activity of rat liver neutral CEase may be under both direct and indirect hormonal control.     

Polyphenols, tannins and antioxidant activities of eight true mangrove plant species in South China

Background and aims

(i) compare the concentrations of total polyphenols (TP) and condensed tannins (CT), and CT profiles in different organs of mature trees and seedlings of eight true mangrove species in Hong Kong; (ii) examine the antioxidant activities of CT and (iii) relate the non-enzymatic antioxidative defence system with the vertical zonation pattern of mangrove species.

Methods

Mature trees and seedlings of eight species were collected from a Hong Kong mangrove swamp to determine TP and CT concentrations and the antioxidant activities of CT.

Results

According to TP concentrations, the true mangrove species could be broadly classified into three groups, (i) Lumnitzera racemosa and Aegiceras corniculatum > (ii) Heritiera littoralis, Excoecaria agallocha, Bruguiera gymnorrhiza and Kandelia obovata > (iii) Acanthus ilicifolius and Avicennia marina. The last two are pioneer species in the most foreshore location. They also had significantly lower antioxidant activities, CT concentrations and different CT profiles than the other six species in mid- and low-tides.

Conclusions

Classification of the eight true mangrove species into three groups based on polyphenols was similar to their vertical zonation from land to sea. The relationships between these antioxidants and zonation should be further verified by transplantation studies.     

The Impact of Industrial Anthropization on Mosquito (Diptera, Culicidae) Communities in Mangrove Areas of Guanabara Bay (Rio de Janeiro, Brazil)

The effects of industrial anthropization on species composition and community diversity of Culicidae (Diptera) were studied in a mangrove area impacted by industrial activities as compared to a preserved area, both around Guanabara Bay in the state of Rio de Janeiro, Brazil. Diversity, equitability, and species richness in Culicidae community differed between the studied areas. Indicator species analysis and correspondence analysis were carried out and indicated that the Sabethini, especially Wyeomyia (Phoniomyia) theobaldi Lane, Wyeomyia (Phoniomyia) fuscipes (Edwards), and a non-identified species of Wyeomyia sp. were associated to the preserved area, whereas Aedes taeniorhynchus Wiedemann and Aedes scapularis (Rondani) to the impacted area.     

High diversity of culturable Burkholderia species associated with sugarcane

Sugarcane is an important crop around the world. Burkholderia genus has emerged as an important plant associated bacteria in the last years. In this study, the occurrence of Burkholderia species associated with two sugarcane varieties cultivated in Mexico was assessed. Burkholderia species were isolated with and without diazotrophs enrichment from sugarcane. Burkholderia strains were identified using a semi-selective set of primers and clustered by restriction analysis of 16S rRNA. The isolates were characterized by 16S rRNA, recA and nifH sequence analysis, whole-cell protein patterns, and plant-growth promotion (PGP) characteristics. Diazotrophic B. unamae and B. tropica were predominant using diazotroph enrichment method. Non-diazotrophic B. cepacia complex (Bcc) species were predominant without enrichment. Among non-diazotrophs, B. tropica was identified. The diazotrophic Burkholderia species exhibit in vitro PGP activities: biosynthesis of indolic compounds, phosphate solubilization, siderophores production and acdS gene presence, which encodes the enzyme ACC (1-aminocyclopropane-1-carboxylate) deaminase. The present study confirms the broad environmental and geographic distribution of diazotrophic B. unamae and B. tropica, and reveals the riches of Bcc and other Burkholderia species associated with sugarcane field-grown in Mexico. This work also shows the potential activities in PGP.     

Comparative study of oxidative stress parameters and acetylcholinesterase activity in the liver of Pelophylax esculentus complex frogs

Comparative activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR), the phase II biotransformation enzyme glutathione-S-transferase (GST), the concentrations of total glutathione (GSH), sulfhydryl groups (–SH) and the activity of the neurotoxicity biomarker acetylcholinesterase (AChE) were investigated in the livers of species belonging to the Pelophylax esculentus “complex” (parental species Pelophylax ridibundus, Pelophylax lessonae, and their hybrid Pelophylax kl. esculentus) from the wetland, Obedska bara in Serbia. The condition factor (CF) and hepato somatic index (HSI) were also calculated. All three species were caught at same locality and were exposed to the same environmental conditions. Liver SOD activity was lower in P. ridibundus than in P. kl. esculentus and P. lessonae; higher activities of CAT, GR and GST were observed in P. kl. esculentus frogs as compared to their parental species. The activity of GSH-Px was significantly lower in P. kl. esculentus. The activity of AChE was increased in P. lessonae as compared to P. kl. esculentus and P. ridibundus. Similar concentrations of GSH and SH groups were observed in all investigated species. P. kl. esculentus had a higher CF, while the HSI was lower when compared to the parental species. Our findings suggest that the parental species (P. ridibundus and P. lessonae) possess more similar antioxidative responses to environmental conditions than the hybrid species P. kl. esculentus. The obtained results improve our understanding of the biology and physiology of these three closely related species.     

Taxus associated fungal endophytes: anticancerous to other biological activities

Endophytes are the plant microorganisms that flourish inside plants (endosphere) and support the growth under normal and challenging environmental circumstances. Besides biological activities, microbial endophytes are important for their contribution in adaptation and survival of the host plants under stressed environments. While various species of Taxus have been globally recognized mainly due to their anticancer property, emphasis has also been on isolation of taxol producing endophytes in view of the taxonomic (endangered) status of the species. Furthermore, Taxus associated endophytes are also being recognized for the biological activities other than taxol production. The present study reviews the diversity of Taxus associated endophytes along with their biological activities. Available literature on Taxus associated endophytes revealed that T. chinensis has received maximum attention (19.5 %), followed by T. wallichiana (18.5 %), T. baccata (16.4 %), T. cuspidata (11.3 %), and T. mairei (10.3 %); with a minimum number of reports accounted for T. globose (1.0 %), and T. celebica (2.0 %). Most fungal endophytes have been identified as ascomycetes. Basidiomycetous endophytes have been reported in a limited number of studies. No endophytes have yet been identified from three species, namely T. controta (West Himalayan yew), T. floridana (Florida yew), and T. canadensis (Canadian yew). A total of 70 diverse endophytic species have been reported from different Taxus species. Fusarium sp. (25.2 %), Aspergillus sp. (17.6 %), Alternaria sp. (11.3 %), Penicillium sp. and Cladosporium sp. (10.0 %) are the five dominant genera. Furthermore, 59.6 % of studies from the reported literature are related to taxol production by fungal endophytes, 9.6 % related to antimicrobial activity, and a combined 15.3 % related to other biological activities, i.e., antifeeding, antimitogenic, antioxidant and cytotoxic activities. Additionally, 3.8 % of fungal endophytes were not studied for any of their biological activities. Only three bacterial endophytes namely Burkholderia sp., Enterobacter sp., and Pseudomonas sp., have been recorded from Taxus along with their contribution in plant growth, antagonistic, and antioxidant activities. Isolation and characterization of microbial endophytes from Taxus plant parts is important for the associated ecology and medicinally important biological activities. It will support the much-required conservation strategy of Taxus species. Further, isolation of plant growth promoting microorganisms from Taxus will help in propagation of the species.     

Catalytic and immunochemical properties of hepatic cytochrome P450 1A in three avian species treated with β-naphthoflavone or isosafrole

Induction of cytochrome P450 1A (CYP1A) can be used as a biomarker of exposure to planar halogenated aromatic hydrocarbons (PHAHs). Our objective was to characterize the induction of CYP1A activity and protein in three avian species following in vivo treatment with β-naphthoflavone (BNF) and/or isosafrole. Alkoxyresorufin-O-dealkylase (alk-ROD) activities of hepatic microsomes from Herring Gulls (Larus argentatus) (HGs), Double-crested Cormorants (Phalacrocorax auritus) (DCCs) and chickens (Gallus domesticus) were measured using ethoxy-, methoxy-, pentoxy- and benzyloxy-resorufin, in the presence and absence of the inhibitors ellipticine or furafylline. Immunoreactivity of microsomal proteins with antibodies to several CYP1A proteins was investigated. CYP1A protein and alk-ROD activities of HGs and DCCs, but not chickens, were induced by isosafrole. Ellipticine was a potent and non-selective inhibitor of alk-ROD activity in all three species, while furafylline inhibition of alk-ROD activities varied among species and treatments. In all three species, BNF induced a protein immunoreactive with monoclonal antibody to CYP1A1 from the marine fish Stenotomus chrysops (scup), but a CYP1A2-like protein was not detected in avian microsomes probed with polyclonal antibodies to mouse CYP1A2. Variations in responses among avian species indicate that CYP1A proteins and substrate specificities should be characterized for each species used in PHAH biomonitoring programs.