Purification and characterization of a novel lipase from the digestive glands of a primitive animal: the scorpion

Higher animal's lipases are well characterized, however, much less is known about lipases from primitive ones. We choose the scorpion, one of the most ancient invertebrates, as a model of a primitive animal. A lipolytic activity was located in the scorpion digestive glands, from which a scorpion digestive lipase (SDL) was purified. Pure SDL, a glycosylated protein, has a molecular mass of 50 kDa, it presents the interfacial activation phenomenon. It was found to be more active on short-chain triacylglycerols than on long-chain triacylglycerols. SDL is a serine enzyme and possesses one accessible sulfhydryl group which is not essential for the catalysis. Among the NH2-terminal 33 residues, a 17 amino acids sequence shows similarities with sequence of Drosophila melanogaster putative lipase. Interestingly, neither colipase, nor bile salts were detected in the scorpion hepatopancreas. This indicates that colipase evolved in vertebrates simultaneously with the appearance of an exocrine pancreas and a true liver which produces bile salts. Furthermore, polyclonal antibodies directed against SDL failed to recognise the classical digestive lipases. Altogether, these results suggest that SDL is a member of a new group of digestive lipases belonging to invertebrates.     

Recruitment of primitive peripheral blood cells: synergism of interleukin 12 with interleukin 6 and stromal cell-derived FACTOR-1

In bone marrow, haematopoietic stem cells (HSC) rely on close contact with stromal cells for proliferation and differentiation. Stromal cell-derived factor (SDF-1) is a chemokine produced by bone marrow stromal cells and has been reported to be a chemoattractant for CD34(+)cells. SDF-1 was evaluated for effects on proliferation of both mature and immature human progenitor cells in vitro. Neither proliferation nor maturation of peripheral blood cells was stimulated by SDF-1 alone. Moreover, we have previously demonstrated that 5-fluorouracile (5-FU) resistant HSC require a combination of interleukin 12 (IL-12), IL-6 and SCF for the production of morphologically recognizable clonogenic elements at day 14 in semisolid medium. Our data reported a strong enhancement of the IL-6, IL-12, SCF-induced synergism (172%) by SDF-1 (296.5%). Furthermore, our data suggest that this chemokine alone had no effect on triggering quiescent cells and may preserve these cells from 5-FU cell damage or upregulate early-acting cytokine receptors. Thus, SDF-1 might play a key role in early human haematopoiesis through its potent synergistic effects in combination with early-acting cytokines. These results suggest that a programmed response to sequential cytokine stimulation may be part of a control mechanism required for maintenance of proliferation of primitive HSC.     

Galactolipase activity of Talaromyces thermophilus lipase on galactolipid micelles,monomolecular films and UV-absorbing surface-coated substrate

Talaromyces thermophilus lipase (TTL) was found to hydrolyze monogalactosyl diacylglycerol (MGDG) and digalactosyl diacylglycerol (DGDG) substrates presented in various forms to the enzyme. Different assay techniques were used for each substrate: pHstat with dioctanoyl galactolipid-bile salt mixed micelles, barostat with dilauroyl galactolipid monomolecular films spread at the air-water interface, and UV absorption using a novel MGDG substrate containing α-eleostearic acid as chromophore and coated on microtiter plates. The kinetic properties of TTL were compared to those of the homologous lipase from Thermomyces lanuginosus (TLL), guinea pig pancreatic lipase-related protein 2 and Fusarium solani cutinase. TTL was found to be the most active galactolipase, with a higher activity on micelles than on monomolecular films or surface-coated MGDG. Nevertheless, the UV absorption assay with coated MGDG was highly sensitive and allowed measuring significant activities with about 10?ng of enzymes, against 100?ng to 10?μg with the pHstat. TTL showed longer lag times than TLL for reaching steady state kinetics of hydrolysis with monomolecular films or surface-coated MGDG. These findings and 3D-modelling of TTL based on the known structure of TLL pointed out to two phenylalanine to leucine substitutions in TTL, that could be responsible for its slower adsorption at lipid-water interface. TTL was found to be more active on MGDG than on DGDG using both galactolipid-bile salt mixed micelles and galactolipid monomolecular films. These later experiments suggest that the second galactose on galactolipid polar head impairs the enzyme adsorption on its aggregated substrate.     

Agricultural wastes as substrates for β-glucosidase production by Talaromyces thermophilus: Role of these enzymes in enhancing waste paper saccharification

In the present study, we investigated a potent extracellular β-glucosidases secreted by the thermophilic fungal strain AX4 of Talaromyces thermophilus, isolated from Tunisian soil samples. This strain was selected referring to the highest thermostability of its β-glucosidases compared to the other fungal isolates. The β-glucosidase production was investigated by submerged fermentation. The optimal temperature and initial pH for maximum β-glucosidase production were 50°C and 7.0, respectively. Several carbon sources were assayed for their effects on β-glucosidase production, significant yields were obtained in media containing lactose 1% (3.0?±?0.36?U/ml) and wheat bran 2% (4.0?±?0.4?U/ml). The combination of wheat bran at 2% and lactose at 0.8% as carbon source enhanced β-glucosidase production, which reached 8.5?±?0.28?U/ml. Furthermore, the β-glucosidase-rich enzymatic juice of T. thermophilus exhibited significant synergism with Trichoderma reesei (Rut C30) cellulases for pretreated waste paper (PWP) hydrolysis. Interestingly, the use of this optimal enzymatic cocktail increased 4.23 fold the glucose yield after saccharification of waste paper. A maximum sugar yield (94%) was reached when using low substrate (2%) and enzyme loading (EC1).     

Endocrine-related reproductive effects in molluscs

Research on endocrine disruption has been a major topic of the past decade. Although most studies concentrated on vertebrate species, invertebrates are now gaining more attention. In particular, data on molluscs is increasing. One of the best-documented and more relevant examples of endocrine disruption is the imposex phenomenon affecting some gastropod species. But the increasing interest is also due to the fact that molluscs, especially bivalves, are good bioindicators used for decades in environmental studies and that progress have been made in the understanding of the physiology and endocrinology of some mollusc species. Recent results suggest that molluscs can be adversely affected by compounds that alter their reproduction and that vertebrate-type sex-steroids metabolism or mechanism of action could be involved in these effects. Nevertheless, the endocrine system of molluscs appears to be dissimilar in many aspects to those of vertebrates and sex-steroids might not have the same importance in all mollusc species. This diversity constitutes an important opportunity to examine and understand new and alternative mechanisms for endocrine disruption.     

Les kystes intraprostatiques: à propos de trois observations

Introduction

Intraprostatic cysts are considered to be rare. Some forms may be responsible for male infertility. The authors study the various clinical aspects, with the aim of defining guidelines for the exploration and treatment of different types of intraprostatic cysts.

Materials and methods

Between 2002 and 2007, we studied 3 cases of intraprostatic cyst in 3 men aged between 32 and 39 years.

Results

The clinical presentation was primary infertility (2 cases) and pelvic pain (2 cases). Digital rectal exam revealed prostate mass in 2 cases. Spermogram results were abnormal in 2 cases. The diagnosis was confirmed by prostate ultrasound. Trans-rectal ultrasound guided needle aspiration was performed in one case with secondary recurrence. Two patients had urethroscopy showing an absence of communication between the cyst and urethral channel. Transurethral incision of the cyst was made with resection of the protruding dome. Progress was favourable in all cases with improvement of semen and dissipation of symptoms without recurrence.

Conclusion

Intraprostatic cysts are benign lesions, often of congenital origin. Imaging is an essential component of diagnosis and can sometimes guide therapeutic procedures. The treatment is not yet codified, but symptomatic and/or complicated cysts can be treated by effective and minimally invasive endoscopic methods.     

A novel hepatopancreatic phospholipase A2 from Hexaplex trunculus with digestive and toxic activities

A marine snail digestive phospholipase A₂ (mSDPL) was purified from delipidated hepatopancreas. Unlike known digestive phospholipases A₂, which are 14 kDa proteins, the purified mSDPL has a molecular mass of about 30 kDa. It has a specific activity of about 180 U/mg measured at 50 °C and pH 8.5 using phosphatidylcholine liposomes as a substrate in the presence of 4 mM NaTDC and 6 mM CaCl₂. The N-terminal amino-acid of the purified mSDPL does not share any homology with known phospholipases.Moreover, the mSDPL exhibits hemolytic activity in intact erythrocytes and can penetrate phospholipid monolayers at high surface pressure, comparable to snake venom PLA₂. These observations suggest that mSDPL could be toxic to mammal cells. However, mSDPL can be classified as a member of a new family of enzymes. It should be situated between the class of toxic phospholipase A₂ from venoms and another class of non toxic pancreatic phospholipase A₂ from mammals.     

Single cell oil production from a newly isolated <Emphasis Type="Italic">Candida viswanathii</Emphasis> Y-E4 and agro-industrial by-products valorization

Microbial lipids have drawn increasing attention in recent years as promising raw materials for biodiesel and added-value compounds production. To this end, new oleaginous yeast, Candida viswanathii Y-E4 was isolated, characterized and used for single cell oil (SCO) production. Physiologic and nutritional parameters optimization was carried out for improved biomass and lipid production. Y-E4 strain was able to use a wide range of substrates, especially C5 and C6 sugars as well as glycerol and hydrophobic substrates. The fatty acid profile analysis showed that oleic acid was the main component produced using different substrates. Batch and fed-bath fermentation were conducted using glucose as carbon source. Lipid production rate is twice higher in fed-batch culture providing a lipid content of 50 % (w/w). To minimize the SCO production cost, C. viswanathii Y-E4 was evaluated for its capacity to use different agro-industrial by-products for microbial oil production and changes in the fatty acid profile were monitored.     

Reuse of Textile Wastewater after Treatment with Isolated Bacteria from Oued Hamdoun River

Industrialization is a boon for developing countries such as Tunisia. However, textile effluents that are being discharged are environmental pollutants, extremely toxic to plants and other living organisms, including humans. The current study was conducted to determine the phytotoxic effect of textile effluents, collected near an industrial zone in the center of Tunisia (Ksar Helal), on the germination and various growth indices of durum wheat (Triticumaestivum L.). Results showed that textile effluent treatments reduced significantly the percentage of seed germination and slowed its kinetic as compared with control. Roots and leaves were also significantly reduced. The phytotoxicity was highly reduced from textile effluents after aerobic biotreatment with bacteria. It can be concluded that the biological treatment process of textile wastewater might serve as a fertilizer production that is able to improve the growth of plants. These results are encouraging in the context of developing a low-budget technology for the effective management of these effluents.     

Trans-glycosylation capacity of a highly glycosylated multi-specific β-glucosidase from Fusarium solani

An extracellular β-glucosidase from Fusaruim solani cultivated on wheat bran was purified by only two chromatographic steps. The purified enzyme exhibited optimal temperature and pH at 60 °C and pH 5, respectively. The purified β-glucosidase behaves as a very large protein due to its high degree of glycosylation. More interestingly, the endoglycosidase H (Endo H) treatment led to 97.55% loss of its initial activity after 24 h of treatment. Besides, the addition of Tunicamycin (nucleoside antibiotic blocking the N-glycosylation first step) during the culture of the fungus affected seriously the glycosylation of the enzyme. Both treatments (endo H and Tunicamycin) strengthened the idea that the hyperglycosylation is involved in the β-glucosidase activity and thermostability. This enzyme was also shown to belong to class III of β-glucosidases (multi-specific) since it was able to act on either cellobiose, gentiobiose or sophorose which are disaccharide composed of two units of d-glucose connected by β1–4, β1–6 and β1–2 linkage, respectively. The β-glucosidase activity was strongly enhanced by ferrous ion (Fe²⁺) and high ionic strength (1 M KCl). The purified enzyme exhibited an efficient transglycosylation capacity allowing the synthesis of cellotriose and cellotetraose using cellobiose as donor.