Structural Insights into the Epimerization of β-1,4-Linked Oligosaccharides Catalyzed by Cellobiose 2-Epimerase, the Sole Enzyme Epimerizing Non-anomeric Hydroxyl Groups of Unmodified Sugars

Cellobiose 2-epimerase (CE) reversibly converts d-glucose residues into d-mannose residues at the reducing end of unmodified β1,4-linked oligosaccharides, including β-1,4-mannobiose, cellobiose, and lactose. CE is responsible for conversion of β1,4-mannobiose to 4-O-β-d-mannosyl-d-glucose in mannan metabolism. However, the detailed catalytic mechanism of CE is unclear due to the lack of structural data in complex with ligands. We determined the crystal structures of halothermophile Rhodothermus marinus CE (RmCE) in complex with substrates/products or intermediate analogs, and its apo form. The structures in complex with the substrates/products indicated that the residues in the β5-β6 loop as well as those in the inner six helices form the catalytic site. Trp-322 and Trp-385 interact with reducing and non-reducing end parts of these ligands, respectively, by stacking interactions. The architecture of the catalytic site also provided insights into the mechanism of reversible epimerization. His-259 abstracts the H2 proton of the d-mannose residue at the reducing end, and consistently forms the cis-enediol intermediate by facilitated depolarization of the 2-OH group mediated by hydrogen bonding interaction with His-200. His-390 subsequently donates the proton to the C2 atom of the intermediate to form a d-glucose residue. The reverse reaction is mediated by these three histidines with the inverse roles of acid/base catalysts. The conformation of cellobiitol demonstrated that the deprotonation/reprotonation step is coupled with rotation of the C2-C3 bond of the open form of the ligand. Moreover, it is postulated that His-390 is closely related to ring opening/closure by transferring a proton between the O5 and O1 atoms of the ligand

Suzaku Observations of PSR B1259-63: A New Manifestation of Relativistic Pulsar Wind

We observed PSR B1259-63, a young non-accreting pulsar orbiting around a Be star SS 2883, eight times with the Suzaku satellite in 2007, to characterize the X-ray emission arising from the interaction between a pulsar relativistic wind and Be star outflows. The X-ray spectra showed a featureless continuum in 0.6-10 keV, modeled by a power law with a wide range of photon index 1.3-1.8. When combined with the Suzaku PIN detector which allowed spectral analysis in the hard 15-50 keV band, X-ray spectra show a break at 5 keV in a certain epoch. Regarding the system as a compactified pulsar wind nebula, in which e+e- pairs are assumed to be accelerated at the inner shock front of the pulsar wind, we attribute the X-ray spectral break to the low-energy cutoff of the synchrotron radiation associated with the Lorentz factor of the relativistic pulsar wind gamma_1 = 4x10^5. Our result indicates that Comptonization of stellar photons by the unshocked pulsar wind will be accessible (or tightly constrained) by observations with the Fermi Gamma-ray Space Telescope during the next periastron passage. The PSR B1259-63 system allows us to probe the fundamental properties of the pulsar wind by a direct means, being complementary to the study of large-scale pulsar wind nebulae.Comment: 11 pages, 9 figures, accepted for publication in The Astrophysical Journa

Pediatric food impaction detected through point-of-care ultrasonography

A previously healthy 1-year-old boy suddenly began coughing while eating a dried sweet potato. Because he continued gagging after vomiting a piece of the dried sweet potato, his mother brought him to our emergency room. Upon arrival, despite normal pulmonary findings, he was drooling and gagging. Ultrasonography was performed because food impaction at the upper esophagus was suspected based on his medical history and physical examination, and results showed that a linear hyperechoic lesion was obstructing the posterior esophageal wall. Computed tomography was also considered for a detailed examination of the foreign body. However, the patient vomited the dried sweet potato before the test was performed. After vomiting the foreign body, the patient stopped gagging, and his ultrasonographic findings were normal. Therefore, the patient was discharged without any complications

Effect of phenol and halogenated phenols on energy transfer reactions of rat liver mitochondria.

The in vitro effects of phenol and p-halogenated phenols on mitochondrial energy transfer reactions were examined using isolated rat liver mitochondria. The relationship between physiochemical properties of phenolic compounds and their effects on mitochondria were studied. Phenol and p-halogenated phenols induced the release of K+ ions from mitochondria, suggesting a change in permeability to K+ ions. A decrease in the respiratory control index, an increase in K+ release and stimulation of latent ATPase activity were observed with these compounds in the descending order of p-iodophenol, p-bromophenol, p-chlorophenol, p-fluorophenol and phenol. The concentrations of the phenolic compounds resulting in fifty percent inhibition of the respiratory control index and those resulting in fifty percent release of K+ ions significantly correlated with Hammett's substituent constant (sigma) and the hydrophobic binding constant (pi) of the compounds.</p