In Vitro Cytotoxicity of Parasporins from Native Algerian Bacillus thuringiensis Strains Against Laryngeal and Alveolar Cancers.

Parasporins (PS), a class of non-insecticidal and non-hemolytic crystal proteins of Bacillus thuringiensis (Bt), are being explored as promising anti-cancer agents due to their specific toxicity to cancer cells. This work is considered as a first initiative aiming at investigating Algerian soil Bt isolates' activity and cytotoxic potential against cancer cells. A total of 48 Bacillus spp. were isolated from different sites in Algeria. Phenotypic and biochemical tests, 16S rDNA molecular identification, and microscopic observation of crystal have confirmed the identification of Bt for ten strains. A screening for non-hemolytic crystalline proteins was performed. Extraction, purification, and activation of non-hemolytic proteins by chromatographic analysis yielded several polypeptides of different molecular weights. A purified PS1, with pro-protein of 81 kDa and several peptides with different molecular weights (18-58 kDa) after activation by trypsin, has been identified from the strain BDzG. The NH2-terminal sequence deciphered in BLAST analysis showed homology to a Bt PS1 protein. Moreover, the screening of parasporin-1 (PS1) gene has also been performed. Cytocidal activity against human epithelial type 2 (HEp2) cells, considered to originate from a human laryngeal carcinoma, was observed with an IC50 equal to 2.33 µg/ml, while moderate cytotoxicity against adenocarcinomic human alveolar basal epithelial (A549) cells has been shown with IC50 equal to 18.54 µg/ml. No cytotoxicity against normal cells was noted. Fluorescence microscopy revealed a condensed or fragmented chromatin indicating the apoptotic death of HEp2 cells. Thus, Bt PS-producer isolated from Algerian soil might have a potential to join the arsenal of natural anti-cancer drugs with high therapeutic potential.

Kinetic study on biodegradation of glyphosate with unacclimated activated sludge.

This article is concerned with the study of biodegradation of an organophosphorus herbicide (glyphosate) using unacclimated activated sludge. Glyphosate at different concentrations (0.1, 0.5, 1, 2 and 5 g/L) was tested for cellular growth. On the other hand, the effect of glyphosate on its own biodegradation was studied by evaluating the fittings of different kinetic models (Andrews, Aiba et al., Han and Levenspiel, Luong, Tessier, Webb, Tseng and Wayman, Yano and Koga). According to the obtained results, the activated sludge was able to use glyphosate as the sole carbon source; however, 2 and 5 g/L glyphosate seemed to inhibit cellular growth. Moreover, glyphosate at initial concentrations of 0.1, 0.5 and 1 g/L was completely degraded within 4, 13 and 18 h of incubation, respectively. Yano and Koga model was the best-fit model (R2 = 0.999, F = 173,106 and P = 0.000006).

Purification and characterization of a highly thermostable chitinase from the stomach of the red scorpionfish Scorpaena scrofa with bioinsecticidal activity toward cowpea weevil Callosobruchus maculatus (Coleoptera: Bruchidae).

This present study is the first attempt to report on the purification and characterization of a chitinase from the stomach of the red scorpionfish Scorpaena scrofa. A 50-kDa chitinase (SsChi50) was purified to homogeneity, and matrix assisted laser desorption ionization-time of flight/mass spectrometry (MALDI-TOF/MS) analysis showed that SsChi50 was a monomer with a molecular mass of 50,103 Da. The 25 N-terminal residues of SsChi50 displayed high homology with family-18 chitinases. Optimal activity was obtained at pH 5.0 at 80 °C. SsChi50 was stable at pH and temperature ranges of 3.0 to 7.0 and 70 to 90 °C for 48 and 4 h respectively. Among the inhibitors and metals tested, p-chloromercuribenzoic acid, N-ethylmaleimide, Hg(2+), and Hg(+) completely inhibited enzyme activity. Chitinase activity was high on colloidal chitin, glycol chitin, glycol chitosane, chitotriose, and chitooligosaccharide. Chitinase activity towards synthetic substrates in the order of p-NP-(GlcNAc)(n) (n = 2-4) was p-NP-(GlcNAc)(2) > p-NP-(GlcNAc)(4) > p-NP-(GlcNAc)(3). Our results suggest that the SsChi50 enzyme preferentially hydrolyzed the second glycosidic link from the non-reducing end of (GlcNAc)(n). This enzyme obeyed Michaelis-Menten kinetics, the K(m) and k(cat) values being 0.412 mg, colloidal chitin mL(-1) and 5.33 s(-1) respectively. An in vivo bioinsecticidal assay was developed for SsChi50 against Callosobruchus maculatus adults. The enzyme showed bioinsecticidal activity toward Callosobruchus maculatus, indicating the possibility of using it in biotechnological strategies for insect management for stored cowpea seeds.

Antagonistic activity of Bacillus sp. obtained from an Algerian oilfield and chemical biocide THPS against sulfate-reducing bacteria consortium inducing corrosion in the oil industry.

The present study enlightens the role of the antagonistic potential of nonpathogenic strain B21 against sulfate-reducing bacteria (SRB) consortium. The inhibitor effects of strain B21 were compared with those of the chemical biocide tetrakishydroxymethylphosphonium sulfate (THPS), generally used in the petroleum industry. The biological inhibitor exhibited much better and effective performance. Growth of SRB in coculture with bacteria strain B21 antagonist exhibited decline in SRB growth, reduction in production of sulfides, with consumption of sulfate. The observed effect seems more important in comparison with the effect caused by the tested biocide (THPS). Strain B21, a dominant facultative aerobic species, has salt growth requirement always above 5% (w/v) salts with optimal concentration of 10-15%. Phylogenetic analysis based on partial 16S rRNA gene sequences showed that strain B21 is a member of the genus Bacillus, being most closely related to Bacillus qingdaonensis DQ115802 (94.0% sequence similarity), Bacillus aidingensis DQ504377 (94.0%), and Bacillus salarius AY667494 (92.2%). Comparative analysis of partial 16S rRNA gene sequence data plus physiological, biochemical, and phenotypic features of the novel isolate and related species of Bacillus indicated that strain B21 may represent a novel species within the genus Bacillus, named Bacillus sp. (EMBL, FR671419). The results of this study indicate the application potential of Bacillus strain B21 as a biocontrol agent to fight corrosion in the oil industry.

Enhancement of the denitrification performance of an activated sludge using an electromagnetic field in batch mode.

The influence of electromagnetic fields on bacterial denitrification has been tested on synthetic media with sludges from wastewater treatment stations, in batch mode. The effects of the intensity of the magnetic induction ratio B (mT), reaction volume and initial biomass concentration on the kinetics of the denitrification process were studied. Magnetic field had both an optimal stimulating effect on the activity of the denitrifying flora for B (mT)/mgx values of the order of 0.212, and an inhibitory effect for the values beyond the latter.Sludges underwent multiple exposure cycles to magnetic fields. It was shown that, after three exposure cycles, denitrification kinetics went from 6.5 to 12.7 mg N-NO-3.L-1.h-1 which corresponds to a 2.7 fold improvement. The improved performance persists even after the cessation of the magnetic field. Observation of the sludge by the environmentalelectron microscope shows that the microbial population forming the starting sludge; changed following exposure to the magnetic field. The action of the; electromagnetic field on the microbial populations in denitrification resulted in the modification of the diversity of the flora that is initially present, favoring the development of Proteo bacteria, particularly the Betaproteo bacteria subclass, which results in improved denitrification.

Membrane bioreactor performance in treating Algiers' landfill leachate from using indigenous bacteria and inoculating with activated sludge.

This study focuses on the treatment of both organic and metallic pollution in the Staoueli landfill leachate. This leachate contains a large amount of organic and inorganic matter and it must imperatively be treated before being released into the environment. Our work presents a comparative study between two membrane sequenced batch bioreactors (B2 contains indigenous leachate bacteria and B1 contains activated sludge). The purpose is to assess the best treatment to use, one that allows the reduction of the polluting load of the leachate and a reduction of membrane fouling. Performances were evaluated by measuring the chemical oxygen demand (COD) and the metal content of the leachate (zinc, iron). The results showed a similar COD removal efficiency in B2 (95%) and B1 (93%). Coupling the bioreactors with an ultrafiltration process allowed a notable reduction in zinc and iron concentrations: Fe of 35% and Zn of 78% for B1UF, and Fe of 71% and Zn of 74% for B2UF.

Anticancer Activity Study of Chromone and Coumarin Hybrids using Electrical Impedance Spectroscopy.

AIMS: Oncology treatments aim at selective toxicity for tumor (compared to normal) cells, and chromone- coumarin hybrids have shown such activity. METHODS: In this study, we test a novel series of synthetic chromone and coumarin derivatives (1-9) for cytotoxic activity against a panel of tumor cell lines (MCF-7, A549, HepG2, HTC-116, B16 and Caco-2) opposed to non-tumor cells (HEK-293t). Electrical impedance spectroscopy was used to monitor cell viability in real time. RESULTS: Compound 8 showed the most potent activity, and it significantly diminished cancer cell proliferation and viability in different cell lines. It induced apoptosis in a dose-dependent manner, as shown by Western blot and flow cytometry. CONCLUSION: Electrical impedance spectroscopy appears to be a convenient tool for in vitro cytotoxicity analysis, which could be useful for identifying drug effects and side effects during early phases of drug discovery and development.

Antimicrobial activity of aqueous extracts from four plants on bacterial isolates from periodontitis patients.

Four aqueous extracts of different plant organs are the following: Artemisia herba-alba, Opuntia ficus-indica, Camellia sinensis and Phlomis crinita were evaluated against two bacterial strains: Porphyromonas gingivalis and Prevotella intermedia, which are implicated in periodontal diseases. By using a disc method, these plant extracts demonstrated powerful bacterial activity against these Gram-negative strains. The minimum inhibitory concentration values of the four plant extracts varied between 0.03 and 590.82 mg/ml for the microbes. Another assay using commercial antibiotics and antibacterials as positive controls was also conducted. Values obtained after statistical analysis of inhibition diameters of all plant extracts demonstrated that for P. gingivalis, the aqueous extracts of A. herba-alba and O. ficus-indica were most effective, followed by those of C. sinensis and P. crinita. For P. intermedia, aqueous extracts of O. ficus-indica and C. sinensis appeared to be more efficient with significantly different (P > 0.05) inhibition diameters, followed by those of O. ficus-indica and P. crinita. In summary, the statistical results reveal that these plant extracts exert stronger antibacterial activity on P. intermedia germ as compared to P. gingivalis.

Anti-proliferative, Cytotoxic and NF-ĸB Inhibitory Properties of Spiro(Lactone-Cyclohexanone) Compounds in Human Leukemia.

BACKGROUND/AIM: NF-ĸB affects most aspects of cellular physiology. Deregulation of NF-ĸB signaling is associated with inflammatory diseases and cancer. In this study, we evaluated the cytotoxic and NF-ĸB inhibition potential of new spiro(lactone-cyclohexanone) compounds in two different human leukemia cell lines (U937 and K562). MATERIALS AND METHODS: The anti-proliferative effects of the spiro(lactone-cyclohexanone) compounds on human K562 and U937 cell lines was evaluated by trypan blue staining, as well as their involvement in NF-kB regulation were analyzed by luciferase reporter gene assay, Caspase-3/7 activities were evaluated to analyze apoptosis induction. RESULTS: Both spiro(coumarin-cyclohexanone) 4 and spiro(6- methyllactone-cyclohexanone) 9 down-regulated cancer cell viability and proliferation. Compound 4 inhibited TNF-α-induced NF-ĸB activation in a dose-dependent manner and induced caspase-dependent apoptosis in both leukemia cell lines. CONCLUSION: Results show that compound 4 and compound 9 have potential as anti-cancer agents. In addition, compound 4 exerted NF-kB inhibition activity in leukemia cancer cells.

Membranes based on polymer miscibility for selective transport and separation of metallic ions.

Polymer inclusion membranes (PIM) used for selective transport and separation of metallic ions have emerged in recent times. Their expansion depends on the method of preparation and their suitable structure and physico-chemical characteristics. In this paper, a novel category of membranes for ions separation is reported. The membranes were synthesized by thermally induced phase separation using a mixture of polyvinylidene fluoride (PVDF) and cellulose triacetate (CTA) plasticized by tris(2-ethylhexyl) phosphate (TEHP) and with di-(2-ethylhexyl) phosphoric acid (D2EHPA) incorporated into the polymer as carrier to increase specific interactions between polymers. PIM membrane exhibited a hydrophobic (∼100°) and thermally stable up to ∼200°C porous homogenous structure. The transport of Ni(II), Zn(II) and Pb(II) from aqueous solutions was studied by competitive transport across polymer inclusion membranes (PIM). Competitive transport of ions in solution across PIM provide the selectivity order: Ni2+ (45%)>Pb2+ (35%)>Zn2+ (5%). A long-term transport experiment was carried out to study the durability of the system.

Purification, characterization, and molecular cloning of an extracellular chitinase from Bacillus licheniformis stain LHH100 isolated from wastewater samples in Algeria.

An extracellular chitinase (ChiA-65) was produced and purified from a newly isolated Bacillus licheniformis LHH100. Pure protein was obtained after heat treatment and ammonium sulphate precipitation followed by Sephacryl S-200 chromatography. Based on matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis, the purified enzyme is a monomer with a molecular mass of 65,195.13 Da. The sequence of the 27 N-terminal residues of the mature ChiA-65 showed high homology with family-18 chitinases. Optimal activity was achieved at pH 4 and 75 °C. Among the inhibitors and metals tested, p-chloromercuribenzoic acid, N-ethylmaleimide, Hg(2+), and Hg(+) completely inhibited enzyme activity. Chitinase activity was high on colloidal chitin, glycol chitin, glycol chitosane, chitotriose, and chitooligosaccharide. Chitinase activity towards synthetic substrates in the order of p-NP-(GlcNAc)n (n = 2-4) was p-NP-(GlcNAc)2 > p-NP-(GlcNAc)4 > p-NP-(GlcNAc)3. Our results suggest that ChiA-65 preferentially hydrolyzed the second glycosidic link from the non-reducing end of (GlcNAc)n. ChiA-65 obeyed Michaelis-Menten kinetics, the Km and kcat values being 0.385 mg, colloidal chitin/ml and 5000 s(-1), respectively. The chiA-65 gene encoding ChiA-65 was cloned in Escherichia coli and its sequence was determined. Above all, ChiA-65 exhibited remarkable biochemical properties suggesting that this enzyme is suitable for bioconversion of chitin waste.

Enzymatic degradation and bioactivity evaluation of C-6 oxidized chitosan.

C-6 oxidized chitosan was produced from chitosan by performing selective oxidation with NaOCl and NaBr using 2,2,6,6-tetramethylpiperidine-1-oxy radical (TEMPO) as catalyst. Endocellulase, Celluclast 1.5 L, Glucanex(®), Macerozyme R-10, hyaluronidase, hyaluronate lyase, red scorpionfish chitinase, glucuronan lyase and a protein mix from Trichoderma reesei were used to degrade the C-6 oxidized chitosan. Glucanex(®), the crude extract from T. reesei IHEM 4122 and Macerozyme R-10 validated the enzymatic degradation through final hydrolysis yields of the derivative respectively close to 36.4, 20.3 and 12.9% (w/w). The best initial reaction velocity (2.41 U/mL) was observed for Glucanex(®). The antileishmanial activity of the derivative was evaluated against Leishmania infantum LIPA 137. The antibacterial activities against Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853 were also tested. Results showed an antileishmanial activity (IC50: 125 µg/mL) of the obtained derivatives against L. infantum LIPA 137.

Equilibrium, kinetic and thermodynamic studies on aluminum biosorption by a mycelial biomass (Streptomyces rimosus).

This work focused on kinetic, equilibrium and thermodynamic studies on aluminum biosorption by Streptomyces rimosus biomass. Infrared spectroscopy analysis shows that S. rimosus present some groups: hydroxyl, methyl, carboxyl, amine, thiol and phosphate. The maximum biosorption capacity of S. rimosus biomass was found to be 11.76 mg g(-1) for the following optimum conditions: particle size, [250-560] µm, pH 4-4.25, biomass content of 25 g L(-1), agitation of 250 rpm and temperature of 25 °C. Langmuir, Freundlich and Dubinin-Radushkevich (D-R) models were applied to describe the biosorption isotherms at free pH (pH(i) 4) and fixed pH (pH(f) 4). Langmuir model is the most adequate. With fixed pH, the maximum biosorption capacity is enhanced from 6.62 mg g(-1) to 11.76 mg g(-1). The thermodynamic parameters (ΔG°, ΔH° and ΔS°) showed the feasibility, endothermic and spontaneous nature of the biosorption at 10-80 °C. The activation energy (Ea) was determined as 52.18 kJ mol(-1) using the Arrhenius equation and the rate constant of pseudo-second-order model (the most adequate kinetic model). The mean free energy was calculated as 12.91 kJ mol(-1) using the D-R isotherm model. The mechanism of Al(III) biosorption on S. rimosus could be a chemical ion exchange and carboxyl groups are mainly involved in this mechanism.