Isolation of algicidal compounds from the red alga <Emphasis Type="Italic">Corallina pilulifera</Emphasis> against red tide microalgae

We determined the structure of two compounds, namely, 5,8,11,14,17-eicosapentaenoic acid (EPA) and di-n-octylphthalate (DnOP), which have algicidal activity against the toxic dinoflagellate, Cochlodinium polykrikoides. The polyunsaturated fatty acid EPA and the anthropogenic DnOP were isolated from the MeOH extract of the red alga Corallina pilulifera. We also found that a commercial EPA has algicidal activity identical to that of the EPA purified from C. pilulifera. At low inoculum (5.0 × 10² cells mL⁻¹), the highest algicidal activity of a commercial EPA exhibited approximately 92.6% algicidal activity after 1 h and 96.8% after 6 h treatment at 6 μg mL⁻¹, respectively. At high inoculum (1.0 × 10⁴ cells mL⁻¹), the strongest algicidal activity of EPA showed 69.5% after 1 h and 75.5% algicidal activity after 6 h treatment at 6 μg mL⁻¹, respectively. However, EPA did not show algicidal activity against several microalgae used in aquaculture such as Pavlova lutheri, Tetraselmis suecica, Isochrysis galbana, and Nannochloris oculata for 6 h treatment at 6 μg mL⁻¹. The algicidal activity of the five red tide strains to EPA (3 μg mL⁻¹) showed about 86.6%, 86.6%, and 67.3% algicidal activity against Skeletonema costatum, Chaetoceros curvisetus, and C. polykrikoides after 1 h treatment at low inoculum (5.0 × 10² cells mL⁻¹), respectively, but not against Prorocentrum minimum and Scrippsiella trochoidea. We concluded that EPA might be useful as a controlling agent of harmful algal blooms.     

Arsenic accumulation and thiol status in lichens exposed to As(V) in controlled conditions

Thalli of epiphytic lichen Hypogymnia physodes (L.) Nyl. and terricolous Cladonia furcata (Huds.) Schrad., collected from an area with background arsenic concentrations, were exposed to 0, 0.1, 1 and 10 μg mL⁻¹ arsenate (As(V)) solutions for 24 h. After exposure they were kept in the metabolically active state for 0, 24 and 48 h in a growth chamber. In the freeze dried samples glutathione (GSH), glutathione disulphide (GSSG), cysteine (Cys) and cystine were analysed and induction of phytochelatin (PC) synthesis measured by reversed-phase high-performance liquid chromatography in combination with fluorescence detection or UV spectrometry. Total arsenic content in thalli was measured by instrumental neutron activation analysis (INAA). In H. physodes, which contained higher amounts of arsenic compared to C. furcata, total glutathione content significantly decreased in samples exposed to 10 μg mL⁻¹ As(V), whereas in C. furcata a significant increase was observed. In both species PC synthesis was induced in thalli exposed to 10 μg mL⁻¹.     

Anticoagulant activity of sulfated polysaccharide isolated from fermented brown seaweed Sargassum fulvellum

A sulfated polysaccharide with anticoagulant properties was isolated from the fermented brown seaweed Sargassum fulvellum. Freeze-dried S. fulvellum was fermented in an incubator for 10^th week at 25°C to convert seaweed macromolecules into anticoagulant sulfated polysaccharides (ASP). Anticoagulant activity was determined by an activated partial thromboplastin time (APTT) test using citrated human blood plasma. The 8^th week S. fulvellum crude seaweed extract (SWE) exhibited the highest blood anticoagulant activity. Therefore, 8^th week crude SWE was used for purification of ASP by two steps; DEAE cellulose anion-exchange followed by Sepharose 4B chromatography. The isolated ASP showed a single spot on agarose gel electrophoresis, which confirmed the purification status of our ASP. Polyacrylamide gel electrophoresis (PAGE) analysis showed that the molecular mass of the purified ASP was between 8 and 20 kDa. Polysaccharide and sulfate concentrations of the purified ASP were 180 and 29.70 μg mL⁻¹ respectively. ASP recovery was 1.32% (w/w) from the crude polysaccharide applied to the DEAE column. Purified ASP had a pH of 3.86 and was considered an acidic polysaccharide. Moreover, both ASP and heparin showed a relative clotting factor of 27.47 at the concentrations of 180 and 60 μg mL⁻¹ respectively. Therefore, S. fulvellum ASP can be considered a weaker anticoagulant than heparin. Results of the APTT, PT, and TT clotting assays showed that ASP was able to inhibit both intrinsic and extrinsic blood coagulation pathways. Finally, this study established a feasible and simple experimental protocol to isolate anticoagulant from fermented seaweeds leading to potential further development of anticoagulant agent for the pharmaceutical industry.     

Identification of carotenoids with high antioxidant capacity produced by extremophile microorganisms

In this study, the carotenoids produced by the extremophile microorganisms Halococcus morrhuae, Halobacterium salinarium and Thermus filiformis were separated and identified by high-performance liquid chromatography connected to a diode array detector and a tandem mass spectrometer. The in vitro scavenging capacity of the carotenoid extracts against radical and non-radical species was evaluated. In halophilic microorganisms, the following carotenoids were identified: bacterioruberin, bisanhydrobacterioruberin, trisanhydrobacterioruberin and their derivatives. In the thermophilic bacterium, the carotenoids all-trans-zeaxanthin, zeaxanthin monoglucoside, thermozeaxanthins and thermobiszeaxanthins were identified. The antioxidant capacities of the carotenoid extracts of H. morrhuae (trolox equivalent antioxidant capacity = 5.07 and IC₅₀ = 0.85 μg mL⁻¹) and H. salinarium (trolox equivalent antioxidant capacity = 5.28 and IC₅₀ = 0.84 μg mL⁻¹) were similar and higher than those of the bacterium T. filiformis (trolox equivalent antioxidant capacity = 2.87 and IC₅₀ = 2.41 μg mL⁻¹). This difference is related to the presence of acyclic carotenoids with both large numbers of conjugated double bounds and of hydroxyl groups in the major carotenoid of the halophilic microorganisms.     

外来红树植物拉关木对乡土种桐花树和正红树的化感作用研究

为了探究外来红树植物拉关木对乡土红树植物的化感作用,该研究观察了不同浓度(0.1、0.3、0.5g·mL~(-1))的拉关木根、叶水浸提液对乡土红树植物桐花树和正红树的胚轴(种子)萌发、幼苗生长及叶片抗氧化酶活性的影响。结果表明:(1)拉关木水浸提液对桐花树种子的成苗率、萌发指数和根长均存在抑制作用,其中对根长的抑制作用随水浸提液浓度的提高而增强。(2)根水浸提液对桐花树幼苗的根长、苗高、生物量等生长指标的影响总体上均表现为低浓度促进,高浓度抑制。(3)拉关木水浸提液对正红树胚轴的萌发率、萌发指数、生长指标均表现为促进作用,且根水浸提液0.1、0.3 g·mL~(-1)处理组的芽长以及根、叶水浸提液0.1、0.3 g·mL~(-1)处理组的生物量显著大于对照组;拉关木水浸提液对正红树幼苗的生物量也表现为促进作用。(4)抗性生理方面,随着拉关木水浸提液浓度的升高,桐花树和正红树幼苗SOD活性降低,正红树幼苗POD活性在根水浸提液0.3 g·mL~(-1)和叶水浸提液0.1 g·mL~(-1)处理组显著高于对照组。以上结果表明,不同乡土植物对拉关木化感作用的敏感性不同,拉关木水浸提液抑制了桐花树的生长,而对正红树的生长则表现出一定程度的促进作用。     

Investigation of Chondrus crispus as a potential source of new antifouling agents

The search for environment-friendly and non-toxic antifouling (AF) paint components has led to the investigation of natural products from seaweeds. The defence metabolites used by algae to deter unwanted epibiosis have potential for harnessing and use in AF applications. Crude algal extracts may provide a suitable mixture of compounds with AF potency. Crude ethanol extracts of the macroalgae Chondrus crispus (Rhodophyceae), from both dried and fresh sources were tested and compared using bioassays based on five marine bacterial strains, five phytoplankton strains and two macroalgae to assess the AF efficacy. Dried extract from the algae had a lower minimum inhibitory concentration at 25 μg mL⁻¹ against the growth of bacteria and phytoplankton species than that from the fresh source. Macroalgae tests indicated that the extracts had an anti-germination activity 25–50 μg mL⁻¹ against both Undaria pinnatifida and Ulva intestinalis spores. A field trial of AF paint incorporating crude extract indicated an initial AF potency lasting six weeks.     

Evaluation of the cytogenetic damage induced by the organophosphorous insecticide acephate

The organophosphorous insecticide acephate was tested for its ability to induce in vitro cytogenetic effect in human peripheral lymphocytes by using the chromosomal aberrations (CAs), sister chromatid exchange (SCE) and micronuclei (MN) assay. The level of nuclear DNA damage of acephate was evaluated by using the comet assay. Concentrations of 12.5, 25, 50, 100 and 200 μg mL⁻¹ of acephate were used. All concentrations of acephate induced significant increase in the frequency of CAs and in the formation of MN dose dependently (r = 0.92 at 24 h, r = 0.95 at 48 h for CAs, r = 0.87 for MN). A significant increase was observed in induction of SCE at 50, 100 and 200 μg mL⁻¹ concentrations during 24 h treatment and at all concentrations (except 12.5 μg mL⁻¹) during 48 h treatment period in a dose-dependent manner (r = 0.84 at 24 h, r = 0.88 at 48 h). Acephate did not affect the replicative index and cytokinesis-block proliferation index (CBPI). However, it significantly decreased the mitotic index at all three highest concentrations (50, 100, 200 μg mL⁻¹) for 24 h treatment and at all concentrations (except 12.5 μg mL⁻¹) for 48 h treatment, dose-dependently (r = 0.94 at 24 h, r = 0.92 at 48 h). A significant increase in mean comet tail length was observed at 100 and 200 μg mL⁻¹ concentrations compared with negative control in a concentration-dependent manner (r = 0.94). The mean comet tail intensity was significantly increased at only 200 μg mL⁻¹ concentration. The present results indicate that acephate is a clastogenic, cytotoxic agent and it causes DNA damage at high concentrations in human lymphocytes in culture.     

Antimicrobial activity of crude extracts from mangrove fungal endophytes

The aim of this work was to select endophytic fungi from mangrove plants that produced antimicrobial substances. Minimal inhibitory concentrations (MIC) and minimal bactericidal concentrations (MBC) or minimal fungicidal concentrations (MFC) of crude extracts from 150 isolates were determined against potential human pathogens by a colorimetric microdilution method. Ninety-two isolates (61.3%) produced inhibitory compounds. Most of the extracts (28–32%) inhibited Staphylococcus aureus (MIC/MBC 4–200/64–200 μg ml⁻¹). Only two extracts inhibited Pseudomonas aeruginosa (MIC/MBC 200/>200 μg ml⁻¹). 25.5 and 11.7% inhibited Microsporum gypseum and Cryptococcus neoformans (MIC/MFC 4–200/8–200 μg ml⁻¹ and 8–200/8–200 μg ml⁻¹, respectively), while 7.5% were active against Candida albicans (MIC/MFC 32–200/32–200 μg ml⁻¹). None of the extracts inhibited Escherichia coli. The most active fungal extracts were from six genera, Acremonium, Diaporthe, Hypoxylon, Pestalotiopsis, Phomopsis, and Xylaria as identified using morphological and molecular methods. Phomopsis sp. MA194 (GU592007, GU592018) isolated from Rhizophora apiculata showed the broadest antimicrobial spectrum with low MIC values of 8–32 μg ml⁻¹against Gram-positive bacteria, yeasts and M. gypseum. It was concluded that endophytic fungi from mangrove plants are diverse, many produce compounds with antimicrobial activity and could be suitable sources of new antimicrobial natural products.     

Effect of macrophyte diet and initial size on the survival and somatic growth of sub-adult <Emphasis Type="Italic">Strongylocentrotus purpuratus</Emphasis>: a laboratory experimental approach

Recently recruited urchins from the same brood, but with different initial sizes, may respond differently to similar environmental factors. The aim of this study was to assess and compare the effects of starvation and diet on the survival, growth rates in size and weight, and gonad index among small and large sub-adult purple sea urchins, Strongylocentrotus purpuratus. Small urchins ranged from 7.3 to 7.8 mm and large urchins from 11.8 to 14.1 mm (test diameters). Two independent experiments were performed. In the first experiment, sea urchins were fed during 22 weeks on Egregia menziesii (ad libitum) and for only 1 day month⁻¹ (starved condition). Feeding regime significantly affected survival, somatic growth rate in size and weight, and gonad index, with higher means in the ad libitum treatments than in starving conditions. A recurrent cannibalism event by conspecifics occurred in small sea urchins under starving conditions. In the second experiment, sea urchins were fed during 13 weeks ad libitum with four diets: kelp (E. menziesii), coralline algae (Bossiella orbigniana), eelgrass (Phyllospadix scouleri) and a mixed diet of the three species. Survival was not affected by diet or urchin size, but diet significantly affected somatic growth rate in size and weight and gonad index. Kelp promoted the highest growth rate (2.23 ± 0.21 mm month⁻¹), the mixed diet produced an intermediate growth (1.26 ± 0.21 mm month⁻¹), while the lowest values corresponded to coralline algae and the eelgrass (0.30 ± 0.12 and 0.10 ± 0.03 mm month⁻¹, respectively, means ± SE). The mean growth rate of small urchins (averaging all diets) was higher than in large specimens (1.17 ± 0.37 and 0.77 ± 0.28 mm month⁻¹, respectively).     

Optimal activity and thermostability of xylose reductase from <Emphasis Type="Italic">Debaryomyces hansenii</Emphasis> UFV-170

Xylose reductase (XR) is the enzyme that catalyzes the first step of xylose metabolism. Although XRs from various yeasts have been characterized, little is known about this enzyme in Debaryomyces hansenii. In the present study, response surface analysis was used to determine the optimal conditions for D. hansenii UFV-170 XR activity. The influence of pH and temperature, ranging from 4.0 to 8.0 and from 25 to 55°C, respectively, was evaluated by a 2² central composite design face-centered. The F-test (ANOVA) and the Student’s t test were performed to evaluate the statistical significance of the model and the regression coefficients, respectively. The NADPH-dependent XR activity varied from 0.502 to 2.53 U mL⁻¹, corresponding to 0.07–0.352 U mg⁻¹, whereas the NADH-dependent one was almost negligible. The model predicted with satisfactory correlation (R ² = 0.940) maximum volumetric activity of 2.27 U mL⁻¹ and specific activity of 0.300 U mg⁻¹ at pH 5.3 and 39°C, which were fairly confirmed by additional tests performed under these conditions. The enzyme proved very stable at low temperature (4°C), keeping its activity almost entirely after 360 min, which corresponded to the half-time at 39°C. On the other hand, at temperatures ≥50°C it was lost almost completely after only 20 min.     

An improved nitrilase-mediated bioprocess for synthesis of nicotinic acid from 3-cyanopyridine with hyperinduced <Emphasis Type="Italic">Nocardia globerula</Emphasis> NHB-2

Nitrilase of Nocardia globerula NHB-2 was induced by short-chain aliphatic nitriles (valeronitrile > isobutyronitrile > butyronitrile > propionitrile) and exhibited activity towards aromatic nitriles (benzonitrile > 3-cyanopyridine > 4-cyanopyridine > m-tolunitrile > p-tolunitrile). Hyperinduction of nitrilase (6.67 U mg_DCW⁻¹, 18.7 U mL⁻¹) was achieved in short incubation time (30 h, 30°C) through multiple feeding of isobutyronitrile in the growth medium. The nitrilase of this organism exhibits both substrate and product inhibition effects. In a fed batch reaction at 1 L scale using hyperinduced resting cells corresponding to 10 U mL⁻¹ nitrilase activity (1.5 mg_DCW mL⁻¹), a total of 123.11 g nicotinic acid was produced at a rate of 24 g h⁻¹ g_DCW⁻¹.     

Mutagenic and Genotoxic Effects of <Emphasis Type="Italic">cis</Emphasis>-(Dichloro)tetraammineruthenium(III) Chloride on Human Peripheral Blood Lymphocytes

Chemotherapeutic agents play an important role in cancer treatment mostly due their systemic action on human organism allowing access to liquid tumors and even metastases. Among these drugs, ruthenium compounds have been showing promising results to treat tumors and represent an important development of new antitumor therapy. This study presents the evaluation of cis-(dichloro)tetraammineruthenium(III) chloride, cis-[RuCl₂(NH₃)₄]Cl, genotoxic effects using human peripheral blood lymphocytes cultured in vitro. Mitotic index (MI), chromosome aberrations (CA), and DNA damage using the comet assay were analyzed. MI in human peripheral blood lymphocyte cultures treated with 1, 10, 100, and 1,000 μg mL⁻¹ cis-[RuCl₂(NH₃)₄]Cl were 5.9%, 4.6%, 3.9%, and 0%, respectively. Doxorubicin chloridate was used as the positive control. CA derived from 1, 10, and 100 μg mL⁻¹ concentrations were defined as spontaneous when compared with the negative control, and at the concentration of 1,000 μg mL⁻¹, the cell cycle was inhibited (IM = 0%). Results obtained for the comet assay using cis-[RuCl₂(NH₃)₄]Cl suggest that this compound has no genotoxic activity against cultured human peripheral blood lymphocytes.     

Growth-inhibiting effects of <Emphasis Type="Italic">Paeonia lactiflora</Emphasis> root steam distillate constituents and structurally related compounds on human intestinal bacteria

The growth-inhibiting activities of Paeonia lactiflora (Paeoniaceae) root steam distillate constituents and structurally related compounds against nine harmful intestinal bacteria and eight lactic acid-producing bacteria were compared with those of two antibiotics, amoxicillin and tetracycline. Thymol, α-terpinolene, (−)-perilla alcohol and (1R)-(−)-myrtenol exhibited high to extremely high levels of growth inhibition of all the harmful bacteria, whereas thymol and α-terpinolene (except for Lactobacillus casei ATCC 393) inhibited the growth of all the beneficial bacteria (MIC, both 0.08–0.62 mg mL⁻¹). Tetracycline and amoxicillin exhibited extremely high level of growth inhibition of all the test bacteria (MIC, <0.00002–0.001 mg mL⁻¹). 1,8-Cineole, geraniol, (−)-borneol, (1S,2S,5S)-(−)-myrtanol, nerol, (S)-(−)-β-citronellol and (±)-lavandulol also exhibited inhibitory activity but with differing specificity and levels of activity. Structure–activity relationship indicates that structural characteristics, such as geometric isomerism, degrees of saturation, types of functional groups and types of carbon skeleton, appear to play a role in determining the growth-inhibiting activity of monoterpenoids. Global efforts to reduce the level of antibiotics justify further studies on naturally occurring P. lactiflora root-derived materials as potential preventive agents against various diseases caused by harmful intestinal bacteria such as clostridia.     

Antioxidant activities in tropical marine macroalgae from the Yucatan Peninsula,Mexico

Extracts from 48 marine macroalgae species (17 Chlorophyta, 8 Phaeophyta and 23 Rhodophyta) from the coasts of Yucatan and Quintana Roo (Mexico) were evaluated for antioxidant activity. The antioxidant activity was measured with the DPPH (2,2-diphenyl-1-picrylhydrasyl) method, and the phenolic content of each extract were also evaluated. All species exhibited a DPPH radical scavenging activity, and three species (Avrainvillea longicaulis, Chondria baileyana and Lobophora variegata) demonstrated great antioxidant potential with very low oxidation index EC₅₀ (1.44 ± 0.01, 2.84 ± 0.07 and 0.32 ± 0.01 mg mL⁻¹, respectively), significantly equivalent to EC₅₀ of some commercial antioxidants such as α-tocopherol, ascorbic acid, BHA and BHT. Moreover, extracts of the most active species exhibited reducing activities, superoxide anion radical scavenging and inhibition of lipid peroxidation. These results suggest that some macroalgae from the Yucatan peninsula have a great antioxidant potential which could be considered for future applications in medicine, food production or cosmetic industry.     

36种中药材体外抗菌活性筛选研究

该文研究36种常用中药材80%乙醇提取物在体外抗临床常见致病菌的抗菌活性。采用药敏纸片法测耐药菌的耐药谱,中药粗粉用80%乙醇浸泡提取,提取液减压浓缩得浸膏,通过琼脂打孔法测定提取物抑菌圈,再通过微量倍比稀释法测定最低抑菌浓度(MIC)和最低杀菌浓度(MBC)。结果表明:36种中药材醇提物中,有15种具有广谱抗菌活性,对实验中各标准菌表现出不同程度的抑制作用,对MRSA抗菌活性也较强。其中,岩陀、卷柏、首乌藤、苏木、乌药、夏枯草6种药材的抗菌活性比较突出,抑菌圈均大于11 mm,细菌对其表现为中高度敏感;它们对7株标准菌的MIC/MBC值除个别为12.5 mg·m L~(-1)以外,均小于1.563 mg·m L~(-1),对16株MRSA的MIC/MBC值均小于1.563 mg·m L~(-1),它们的萃取层活性均小于1 mg·m L~(-1)。所筛选出的15种抗菌活性较强的中药材,可为后续研究其活性单体化合物和作用机制,研发有效的抗多重耐药菌的中药制剂以及解决细菌耐药性问题提供一定的参考。     

Effects of salts on activity of halophilic cellulase with glucomannanase activity isolated from alkaliphilic and halophilic Bacillus sp. BG-CS10

Alkaliphilic and halophilic Bacillus sp. BG-CS10 was isolated from Zabuye Salt Lake, Tibet. The gene celB, encoding a halophilic cellulase was identified from the genomic library of BG-CS10. CelB belongs to the cellulase superfamily and DUF291 superfamily, with an unknown function domain and less than 58% identity to other cellulases in GenBank. The purified recombinant protein (molecular weight: 62 kDa) can hydrolyze soluble cellulose substrates containing beta-1,4-linkages, such as carboxylmethyl cellulose and konjac glucomannan, but has no exoglucanase and β-glucosidase activities. Thus, CelB is a cellulase with an endo mode of action and glucomannanase activity. Interestingly, the enzyme activity was increased approximately tenfold with 2.5 M NaCl or 3 M KCl. Furthermore, the optimal temperatures were 55°C with 2.5 M NaCl and 35°C without NaCl, respectively. This indicates that NaCl can improve enzyme thermostability. The K _m and k _cat values of CelB for CMC with 2.5 M NaCl were 3.18 mg mL⁻¹ and 26 s⁻¹, while the K _m and k _cat values of CelB without NaCl were 6.6 mg mL⁻¹ and 2.1 s⁻¹. Thus, this thermo-stable, salt and pH-tolerant cellulase is a promising candidate for industrial applications, and provides a new model to study salt effects on the structure of protein.     

Bioremoval of hexavalent chromium from water by a salt tolerant bacterium, Exiguobacterium sp. GS1

Pollution of terrestrial surfaces and aquatic systems by hexavalent chromium, Cr(VI), is a worldwide public health problem. A chromium resistant bacterial isolate identified as Exiguobacterium sp. GS1 by 16S rRNA gene sequencing displayed high rate of removal of Cr(VI) from water. Exiguobacterium sp. GS1 is 99% identical to Exiguobacterium acetylicum. The isolate significantly removed Cr(VI) at both high and low concentrations (1–200 μg mL⁻¹) within 12 h. The Michaelis–Menten K _m and V _max for Cr(VI) bioremoval were calculated to be 141.92 μg mL⁻¹ and 13.22 μg mL⁻¹ h⁻¹, respectively. Growth of Exiguobacterium sp. GS1 was indifferent at 1–75 μg mL⁻¹ Cr(VI) in 12 h. At initial concentration of 8,000 μg L⁻¹, Exiguobacterium sp. GS1 displayed rapid bioremoval of Cr(VI) with over 50% bioremoval in 3 h and 91% bioremoval in 8 h. Kinetic analysis of Cr(VI) bioremoval rate revealed zero-order in 8 h. Exiguobacterium sp. GS1 grew and significantly reduced Cr(VI) in cultures containing 1–9% salt indicating high salt tolerance. Similarly the isolate substantially reduced Cr(VI) over a wide range of temperature (18–45  °C) and initial pH (6.0–9.0). The T _opt and initial pH_opt were 35–40  °C and 7–8, respectively. Exiguobacterium sp. GS1 displayed a great potential for bioremediation of Cr(VI) in diverse complex environments.     

Solid state bioreactor production of transglutaminase by Amazonian Bacillus circulans BL32 strain

In this work, we investigated the production of transglutaminase (TGase) by an Amazonian isolated strain of Bacillus circulans by solid-state cultivation (SSC). Several agro-industrial residues, such as untreated corn grits, milled brewers rice, industrial fibrous soy residue, soy hull, and malt bagasse, were used as substrates for microbial growth and enzyme production. Growth on industrial fibrous soy residue, which is rich in protein and hemicellulose, produced the highest TGase activity (0.74 U g⁻¹ of dried substrate after 48 h of incubation). A 2³ central composite design was applied to determine the optimal conditions of aeration, cultivation temperature and inoculum cell concentration to TGase production. The best culture conditions were determined as being 0.6 L air min⁻¹, 33 °C and 10 log ₁₀ CFU g⁻¹ of dried substrate, respectively. Under the proposed optimized conditions, the model predicted an enzyme production of 1.16 U g⁻¹ of dried substrate, closely matching the experimental activity of 1.25 U g⁻¹. Results presented in this work point to the use of this newly isolated B. circulans strain as a potential alternative of microbial source for TGase production by SSC, using inexpensive culture media.     

Anticoagulant activity, paw edema and pleurisy induced carrageenan: Action of major types of commercial carrageenans

Considering the use of commercials carrageenans as a model for inflammation, the aim this work is a comparative study these compounds in the pro-inflammatory action by ear edema and pleurisy and to analyze their anticoagulant activity. Paw edema was induced by injecting kappa, iota and lambda carrageenans in saline solution into the hind paw of male Wistar rats (p < 0.05). The three types of carrageenans showed different volume in pleural exudates, characterized by fluid accumulation, a large number of neutrophils and raised NO production (p < 0.001). The activated partial thromboplastin time (aPTT) for kappa and iota carrageenans (100 μg) was 240 s and 132 s, respectively. Lambda carrageenan was the most potent anticoagulant at 240 s (20 μg). These carragenans demonstrated no anticoagulant action using the PT test in vitro. Histological analysis in paw edema demonstrated that iota and lambda carrageenans showed major cellular infiltration in relation to kappa. Thus, quantitative evaluation of inflammation demonstrated that iota and lambda carrageenans have higher inflammatory potential than do kappa carrageenans.     

Antiparkinsonian Effects of Aqueous Methanolic Extract of Hyoscyamus niger Seeds Result From its Monoamine Oxidase Inhibitory and Hydroxyl Radical Scavenging Potency

Hyoscyamus species is one of the four plants used in Ayurveda for the treatment of Parkinson’s disease (PD). Since Hyoscyamus niger was found to contain negligible levels of L-DOPA, we evaluated neuroprotective potential, if any, of characterized petroleum ether and aqueous methanol extracts of its seeds in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model of PD in mice. Air dried authenticated H. niger seeds were sequentially extracted using petroleum ether and aqueous methanol and were characterized employing HPLC-electrochemistry and LCMS. Parkinsonian mice were treated daily twice with the extracts (125–500 mg/kg, p.o.) for two days and motor functions and striatal dopamine levels were assayed. Administration of the aqueous methanol extract (containing 0.03% w/w of L-DOPA), but not petroleum ether extract, significantly attenuated motor disabilities (akinesia, catalepsy and reduced swim score) and striatal dopamine loss in MPTP treated mice. Since the extract caused significant inhibition of monoamine oxidase activity and attenuated 1-methyl-4-phenyl pyridinium (MPP⁺)-induced hydroxyl radical (·OH) generation in isolated mitochondria, it is possible that the methanolic extract of Hyoscyamus niger seeds protects against parkinsonism in mice by means of its ability to inhibit increased ·OH generated in the mitochondria.