Hyper‐production of α‐amylase from agro‐residual medium with high‐glucose in SSF using catabolite derepressed Bacillus subtilis KCC103

In Bacillus subtilis KCC103, α‐amylase is hyper‐produced and α‐amylase synthesis is not subject to catabolite repression. The α‐amylase was produced from KCC103 by solid‐state fermentation (SSF) using agro‐residues and oil cakes as growth substrates. The KCC103 was also tested for its resistance to repression by hyper level (>10% w/w) of glucose and xylose on α‐amylase production in SSF. Among growth media containing various combinations of agro‐residues, the medium with wheat bran and sunflower oil cake supported highest enzyme production (20700 IU (g dry wt)^–1). The α‐amylase production was enhanced (4.2 folds) by optimizing the growth substrate and the process parameters: the optimal conditions were wheat bran:sun flower oil cake ratio‐1:1 (w/w), substrate particle size‐500 μm, substrate to flask volume‐1:100 (w/v), initial substrate moisture content‐90% (v/w), inoculum size‐35%, initial medium pH‐7.0, growth temperature‐37 °C and cultivation time‐48 h. α‐Amylase production was further enhanced up to 1.7 folds when SSF was carried out using optimized medium supplemented with sugars or yeast extract (1% w/v) under optimized conditions. Supplementation of biomass sugars, glucose or xylose at 20% (w/w), did not repress the synthesis of α‐amylase showing the hyper‐tolerance of KCC103 to repression by simple sugars on α‐amylase production in SSF. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Antibacterial and Anticancer activity of ε ‐poly‐L‐lysine (ε ‐PL) produced by a marine Bacillus subtilis sp.

A marine Bacillus subtilis SDNS was isolated from sea water in Alexandria and identified using 16S rDNA sequence analysis. The bacterium produced a compound active against a number of gram negativeve bacteria. Moreover, the anticancer activity of this bacterium was tested against three different human cell lines (Hela S3, HepG2 and CaCo). The highest inhibition activity was recorded against Hela S3 cell line (77.2%), while almost no activity was recorded towards CaCo cell line. HPLC and TLC analyses supported evidence that Bacillus subtilis SDNS product is ?;‐poly‐L‐lysine. To achieve maximum production, Plackett‐Burman experimental design was applied. A 1.5 fold increase was observed when Bacillus subtilis SDNS was grown in optimized medium composed of g/l: (NH₄)₂SO₄, 15; K₂HPO₄, 0.3; KH₂PO₄, 2; MgSO₄ · 7 H₂O, 1; ZnSO₄ · 7 H₂O, 0; FeSO₄ · 7 H₂O, 0.03; glucose, 25; yeast extract, 1, pH 6.8. Under optimized culture condition, a product value of 76.3 mg/l could be obtained. According to available literature, this is the first announcement for the production of ?;‐poly‐L‐lysine (?;‐PL) by a member of genus Bacillus. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Salivary α‐amylase stress reactivity across different age groups

Salivary α‐amylase (sAA) increases rapidly in response to psychosocial stress in young adults, but no direct comparisons between different age groups across the life span have been made. Secretion of sAA and cortisol was assessed in children, young adults, and older adults after exposure to the Trier Social Stress Test. Additionally, cardiovascular activity was measured in both adult groups. Older adults showed attenuated sAA, heart rate (HR), and heart rate variability (HRV) responses. Furthermore, we found higher sAA but lower cortisol at baseline as well as lower sAA and cortisol responses in children. Age × sex interactions were observed only for cortisol with higher responses in older male participants. No associations between the parameters were found. These results implicate sAA as an alternative or additional sympathetic stress marker throughout the life span, with marked and rapid stress responsiveness in three relevant age groups.     

Synthesis and Micellization Properties of Novel Symmetrical Poly(ε‐caprolactone‐b‐[R,S] β‐malic acid‐b‐ε‐caprolactone) Triblock Copolymers

Summary: A four‐step strategy to synthesize well‐defined amphiphilic poly(ε‐caprolactone‐b‐[R,S] β‐malic acid‐b‐ε‐caprolactone) triblock copolymers [P(CL‐b‐MLA‐b‐CL)], which combines the anionic polymerization of [R,S] benzyl β‐malolactonate (MLABz), and the coordination‐insertion ring‐opening polymerization (ROP) of ε‐caprolactone (CL), followed by the selective removal of benzyloxy protective groups of the central poly(malolactonate) block is described. The first step involves MLABz initiated by potassium 11‐hydroxydodecanoate in the presence of 18‐crown‐6 ether. This step was carried out at 0 °C with an initial monomer concentration of 0.2 mol · L^?1 in order to limit the occurrence of undesirable transfer and termination reactions by proton abstraction. After selective reduction of the carboxylic acid end‐group of the resulting α‐hydroxy, ω‐carboxylic poly([R,S] benzyl β‐malolactonate) leading to an α,ω‐dihydroxy PMLABz, the polymerization of CL was initiated by each hydroxyl end‐groups previously activated by AlEt₃. Finally, after catalytic hydrogenation of the benzyl ester functions, the P(CL‐b‐MLA‐b‐CL) triblock copolymer was recovered and the amphiphilic character evidenced by UV spectroscopy. As demonstrated, the CMC of these new P(CL‐b‐MLA‐b‐CL) triblock copolymer is higher by one order of magnitude than that of a P(MLA‐b‐CL) diblock copolymer of similar composition.

Concentration dependence of pyrene I₃₃₈/I₃₃₅ intensitiy ratio for P(MLA‐b‐CL) diblock and P(CL‐b‐MLA‐b‐CL) triblock copolymers in water.     

Expression analysis of an evolutionary conserved metallophosphodiesterase gene,Mpped1, in the normal and β‐catenin–deficient malformed dorsal telencephalon

We report the expression of the mouse Mpped1 in the telencephalon through embryonic stages to adulthood. Using Northern blotting analysis and RNA in situ hybridization (ISH), our data show that Mpped1 is specifically expressed in the brain and is enriched in the cortical plate of the developing telencephalon. Postnatally, the expression of Mpped1 is reduced in the cerebral cortex relative to its levels in the embryonic dorsal telencephalon. Also, Mpped1 expression is sustained in the hippocampal CA1 region. Examination of the expression of Mpped1 and other cortical layer markers by ISH in a malformed β‐catenin null dorsal telencephalon show that the Mpped1‐, Cux2‐, and Rorβ‐expressing superficial cortical layers are reduced and form patchy patterns, and the Tbr‐1‐expressing deep‐layer neurons are incorrectly located on superficial layers, indicative of a migration defect of cortical neurons in the absence of β‐catenin. Developmental Dynamics 239:1797–1806, 2010. © 2010 Wiley‐Liss, Inc.     

ORIGINAL ARTICLE: Expression of IL‐6, IL‐8, TNF‐α, IL‐10, HSP‐60, Anti‐HSP‐60 Antibodies,and Anti‐sperm Antibodies,in Semen of Men with Leukocytes and/or Bacteria

Citation Martínez‐Prado E, Bermúdez MIC. Expression of IL‐6, IL‐8, TNF‐α, IL‐10, HSP‐60, anti‐HSP‐60 antibodies, and anti‐sperm antibodies, in semen of men with leukocytes and/or bacteria. Am J Reprod Immunol 2010; 63: 233–243 Problem Different cellular and biochemical markers have been proposed as indicators of infection‐inflammation of male genital tract. Method of study Semen samples from 80 men attending an andrologic clinic were evaluated to determine the presence of leukocyte, bacteria, antibodies against Chlamydia trachomatis, levels of IL‐6, IL‐8, IL‐10, and TNF‐α, HSP‐60, anti‐HSP‐60 antibodies, and anti‐sperm antibodies. Results Leukocytes in semen significantly correlated with an increase in IL‐6, IL‐8, and TNF‐α. The simultaneous presence of pathogens and leukocytes was associated with high levels of IL‐8 and TNF‐α, whereas IL‐6 was more associated with the presence of leukocytes. Anti‐HSP‐60 antibodies positively correlated with IL‐6 and IL‐8. The presence of anti‐sperm antibodies highly associated with an increase in anti‐HSP‐60 antibodies. Conclusions The type of cytokines present in the semen will depend on the single or simultaneous presence of leukocytes and/or pathogens. Chronic male genital tract infections could be associated with the development of anti‐HSP‐60 antibodies and anti‐sperm antibodies.