Catalysis by hen egg-white lysozyme proceeds via a covalent intermediate

Hen egg-white lysozyme (HEWL) was the first enzyme to have its three-dimensional structure determined by X-ray diffraction techniques(1). A catalytic mechanism, featuring a long-lived oxo-carbenium-ion intermediate, was proposed on the basis of model-building studies(2). The `Phillips' mechanism is widely held as the paradigm for the catalytic mechanism of beta -glycosidases that cleave glycosidic linkages with net retention of configuration of the anomeric centre. Studies with other retaining beta -glycosidases, however, provide strong evidence pointing to a common mechanism for these enzymes that involves a covalent glycosyl-enzyme intermediate, as previously postulated(3). Here we show, in three different cases using electrospray ionization mass spectrometry, a catalytically competent covalent glycosyl-enzyme intermediate during the catalytic cycle of HEWL. We also show the three-dimensional structure of this intermediate as determined by Xray diffraction. We formulate a general catalytic mechanism for all retaining beta -glycosidases that includes substrate distortion, formation of a covalent intermediate, and the electrophilic migration of C1 along the reaction coordinate

Polyunsaturated fatty acid intake and prevalence of eczema and rhinoconjunctivitis in Japanese children: The Ryukyus Child Health Study

<p>Abstract</p> <p>Background</p> <p>The recent increase in the prevalence of allergic disorders might be a consequence of increased intake of n-6 polyunsaturated fatty acids (PUFAs) and reduced intake of n-3 PUFAs. The current cross-sectional study examined the association between intake levels and the prevalence of eczema and rhinoconjunctivitis in Japanese children.</p> <p>Methods</p> <p>Subjects were 23,388 schoolchildren aged 6-15 years residing in Okinawa. The presence of eczema and/or rhinoconjunctivitis was determined according to the criteria of the International Study of Asthma and Allergies in Childhood. A brief diet history questionnaire for children and adolescents was administered to acquire information on dietary factors. Adjustment was made for age, sex, residential municipality, number of siblings, smoking in the household, body mass index, paternal and maternal history of allergic diseases, and paternal and maternal educational level.</p> <p>Results</p> <p>The prevalences of eczema and rhinoconjunctivitis in the previous 12 months were 7.0% and 8.0%, respectively. Consumption of PUFAs, n-3 PUFAs, α-linolenic acid, n-6 PUFAs, and linoleic acid was positively associated with the prevalence of eczema: the adjusted odds ratios (ORs) between extreme quintiles (95% confidence intervals [CIs], <it>P </it>for trend) were 1.26 (1.07-1.48, 0.04), 1.31 (1.11-1.54, 0.009), 1.31 (1.12-1.55, 0.003), 1.26 (1.07-1.48, 0.01), and 1.27 (1.08-1.49, 0.01), respectively. Arachidonic acid intake was independently inversely related to eczema: the adjusted OR between extreme quintiles was 0.81 (0.69-0.95, 0.0008). Eczema was not associated with eicosapentaenoic or docosahexaenoic acid intake, or with the ratio of n-3 to n-6 PUFA intake. Only arachidonic acid intake was statistically significantly related to the prevalence of rhinoconjunctivitis, showing a clear inverse linear trend: the adjusted OR between extreme quintiles was 0.86 (0.74-0.997, 0.03).</p> <p>Conclusions</p> <p>Consumption of n-3 and n-6 PUFAs, especially α-linolenic acid and linoleic acid, may be positively associated with eczema. Arachidonic acid intake may be inversely related to eczema and rhinoconjunctivitis.</p

Aquifers and hyporheic zones: Towards an ecological understanding of groundwater

Ecological constraints in subsurface environments relate directly to groundwater flow, hydraulic conductivity, interstitial biogeochemistry, pore size, and hydrological linkages to adjacent aquifers and surface ecosystems. Groundwater ecology has evolved from a science describing the unique subterranean biota to its current form emphasising multidisciplinary studies that integrate hydrogeology and ecology. This multidisciplinary approach seeks to elucidate the function of groundwater ecosystems and their roles in maintaining subterranean and surface water quality. In aquifer-surface water ecotones, geochemical gradients and microbial biofilms mediate transformations of water chemistry. Subsurface fauna (stygofauna) graze biofilms, alter interstitial pore size through their movement, and physically transport material through the groundwater environment. Further, changes in their populations provide signals of declining water quality. Better integrating groundwater ecology, biogeochemistry, and hydrogeology will significantly advance our understanding of subterranean ecosystems, especially in terms of bioremediation of contaminated groundwaters, maintenance or improvement of surface water quality in groundwater-dependent ecosystems, and improved protection of groundwater habitats during the extraction of natural resources. Overall, this will lead to a better understanding of the implications of groundwater hydrology and aquifer geology to distributions of subsurface fauna and microbiota, ecological processes such as carbon cycling, and sustainable groundwater management