Transglycosylation Activity of Engineered Bifidobacterium Lacto-N-Biosidase Mutants at Donor Subsites for Lacto-N-Tetraose Synthesis

The health benefits of human milk oligosaccharides (HMOs) make them attractive targets as supplements for infant formula milks. However, HMO synthesis is still challenging and only two HMOs have been marketed. Engineering glycoside hydrolases into transglycosylases may provide biocatalytic routes to the synthesis of complex oligosaccharides. Lacto-N-biosidase from Bifidobacterium bifidum (LnbB) is a GH20 enzyme present in the gut microbiota of breast-fed infants that hydrolyzes lacto-N-tetraose (LNT), the core structure of the most abundant type I HMOs. Here we report a mutational study in the donor subsites of the substrate binding cleft with the aim of reducing hydrolytic activity and conferring transglycosylation activity for the synthesis of LNT from p-nitrophenyl β-lacto-N-bioside and lactose. As compared with the wt enzyme with negligible transglycosylation activity, mutants with residual hydrolase activity within 0.05% to 1.6% of the wild-type enzyme result in transglycosylating enzymes with LNT yields in the range of 10–30%. Mutations of Trp394, located in subsite -1 next to the catalytic residues, have a large impact on the transglycosylation/hydrolysis ratio, with W394F being the best mutant as a biocatalyst producing LNT at 32% yield. It is the first reported transglycosylating LnbB enzyme variant, amenable to further engineering for practical enzymatic synthesis of LNT.     

Perceived stress management skill mediates the relationship between optimism and positive mood following radical prostatectomy.

This study evaluated relations among optimism, perceived stress management skills (PSMS), and positive mood in 46 men who had surgical treatment for localized prostate cancer. The authors found that optimism, PSMS, and positive mood scores were positively correlated. Positive mood was unrelated to demographic and disease-related control variables. In a hierarchical regression model controlling for PSMS, the relationship between optimism and positive mood became nonsignificant, whereas PSMS remained a correlate of positive mood. Results suggest that the relationship between optimism and positive mood may be mediated by belief in being able to use stress management techniques effectively. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Deorthogonalization of ODS-CDMA QPSK by AM/PM Nonlinearity

When using a conventional demodulator, a sufficient condition to maintain ODS-CDMA codeword orthogonality is constant relative channel amplitude over the codeword duration. When transmitted over a memoryless, AM/PM nonlinearity channel, the chip-to-chip fluctuating amplitude of a composite ODS-CDMA QPSK waveform produces a chip-to-chip fluctuating phase that deorthogonalizes the ODS-CDMA codewords, resulting in an additional multiple access self-interference similar to that found in asynchronous DS-CDMA. For ODS-CDMA QPSK in an AM/PM nonlinearity channel, we utilize the Central Limit Theorem, derive, and evaluate: (i) an expression for the signal-to-distortion ratio at the demodulator output, (ii) an expression for the uncoded bit error probability, and (iii) an upper bound on the convolutionally-coded bit error probability. We find that the degradation to BER depends on both the AM/PM nonlinearity slope and the ODS-CDMA channel loading.     

Sustainability: ecological, social, economic, technological, and systems perspectives

Sustainability is generally associated with a definition by the World Commission on Environment and Development, 1987: "... development that meets the needs of the present without compromising the ability of future generations to meet their own needs ..." However, there is no mathematical theory embodying these concepts, although one would be immensely valuable in humanity's efforts to manage the environment. The concept of sustainability applies to integrated systems comprising humans and the rest of nature; the structures and operation of the human component (society, economy, law, etc.) must be such that they reinforce the persistence of the structures and operation of the natural component (ecosystem trophic linkages, biodiversity, biogeochemical cycles, etc.). One of the challenges of sustainability research lies in linking measures of ecosystem functioning to the structure and operation of the associated social system. We review the nature of this complex system including its ecological, social, economic, and technological aspects, and propose an approach to assessing sustainability based on Information Theory that bridges the natural and human systems. These principles are then illustrated using a model system with an ecological food web linked to a rudimentary social system. This work is part of the efforts of a larger multidisciplinary group at the U.S. Environmental Protection Agency's National Risk Management Research Laboratory.     

Defensiveness, trait anxiety, and Epstein-Barr viral capsid antigen antibody titers in healthy college students.

Investigated the relationship of individual differences in repressive coping styles with differences in antibody titer to Epstein-Barr (EB) viral capsid antigen in a normal, healthy college population made up of people previously exposed to EB. Each of 54 1st-yr undergraduates completed a battery of physical-status questions and items pertaining to potential behavioral immunomodulatory confounds, along with the Taylor Manifest Anxiety Scale and the Marlowe-Crowne Social Desirability Scale. Ss reporting high and middle levels of anxiety had higher antibody titers to EB, suggesting poorer immune control over the latent virus, as compared with the low-anxious group. Similarly, high-defensive Ss had higher antibody titers than their low-defensive counterparts, and neither group differed from the middle group. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Subcellular Localization of Acyl-CoA: Lysophosphatidylethanolamine Acyltransferases (LPEATs) and the Effects of Knocking-Out and Overexpression of Their Genes on Autophagy Markers Level and Life Span of A. thaliana

Arabidopsis thaliana possesses two acyl-CoA:lysophosphatidylethanolamine acyltransferases, LPEAT1 and LPEAT2, which are encoded by At1g80950 and At2g45670 genes, respectively. Both single lpeat2 mutant and double lpeat1 lpeat2 mutant plants exhibit a variety of conspicuous phenotypes, including dwarfed growth. Confocal microscopic analysis of tobacco suspension-cultured cells transiently transformed with green fluorescent protein-tagged versions of LPEAT1 or LPEAT2 revealed that LPEAT1 is localized to the endoplasmic reticulum (ER), whereas LPEAT2 is localized to both Golgi and late endosomes. Considering that the primary product of the reaction catalyzed by LPEATs is phosphatidylethanolamine, which is known to be covalently conjugated with autophagy-related protein ATG8 during a key step of the formation of autophagosomes, we investigated the requirements for LPEATs to engage in autophagic activity in Arabidopsis. Knocking out of either or both LPEAT genes led to enhanced accumulation of the autophagic adaptor protein NBR1 and decreased levels of both ATG8a mRNA and total ATG8 protein. Moreover, we detected significantly fewer membrane objects in the vacuoles of lpeat1 lpeat2 double mutant mesophyll cells than in vacuoles of control plants. However, contrary to what has been reported on autophagy deficient plants, the lpeat mutants displayed a prolonged life span compared to wild type, including delayed senescence.