Engineering a twist in 9,10-diethynylanthracenes by steric interactions.

A series of 9,10-bis(phenylethynyl)anthracenes decorated with sterically demanding tert-butyl substituents have been prepared and spectroscopically characterised. We demonstrate that the introduction of two bulky substituents in the ortho position of the phenyl rings effectively locks the ground state into a conformation in which the three rings are orthogonal. Fluorescence spectroscopy reveals evidence for partial planarisation of this compound in the excited state at ambient temperature, but this is prevented in low temperature solvent glasses.     

The effects of depowered airbags on skin injuries in frontal automobile crashes

The purpose of this study was to determine the effects of depowered frontal airbags on the incidence of skin injuries. The National Automotive Sampling System database files from 1993 to 2000 were examined in a study including 2,246,524 occupants exposed to airbag deployment in the United States. There was no significant difference between full-powered and depowered airbags, with 60.2 percent of those exposed to a full-powered deployment sustaining a skin injury versus 59.5 percent of occupants exposed to a depowered airbag (p = 0.19). Whether occupants were exposed to a full-powered airbag (1,936,485 occupants) or a depowered airbay (310,039 occupants), the majority of skin injuries were to the upper extremity and the face. Regardless of airbag power, the overwhelming majority of the skin injuries were minor (99.8 percent). There was not a significantly greater risk of injury from any source for occupants exposed to a depowered airbag or a full-powered airbag (p = 0.87). The data suggest that the implementation of depowered airbags did not affect the number, seriousness, location, or source of skin injuries.     

Improved productivity of <Emphasis Type="Italic">β</Emphasis>-fructofuranosidase by a derepressed mutant of <Emphasis Type="Italic">Aspergillus niger</Emphasis> from conventional and non-conventional substrates

Summary Wild-type cultures of Aspergillus niger produced a basal level of β-fructofuranosidase on glucose of 1 IU l⁻¹ h⁻¹. In contrast, a catabolite-derepressed mutant strain of the same organism produced a markedly higher level (25 IU l⁻¹ h⁻¹) of this enzyme when grown on the same carbon source. Wheat bran induced both the wild type (252 IU l⁻¹ h⁻¹) and the mutant strain (516 IU l⁻¹ h⁻¹) to produce 252- to 516-fold higher levels of this enzyme than was observed with the wild-type grown on glucose and was the best carbon source. When corn steep liquor served as a nitrogen source, the wild-type organism showed a higher activity of enzyme on monosaccharides and disaccharides comparable to that produced by corncobs in the basal medium and that mutant was a potentially improved (> 2-fold) organism for the production of β-fructofuranosidase on all carbon sources. Enhanced substrate consumption and product formation kinetic parameters suggest that the mutant organism may be exploited for bulk production of this useful enzyme.     

Nitrogen availability of anaerobic swine lagoon sludge: sludge source effects

Increased numbers of swine producers will be removing sludge from their anaerobic waste treatment lagoons in the next few years, due to sludge exceeding designed storage capacity. Information on availability of nitrogen (N) in the sludge is needed to improve application recommendations for crops. The objective of this study was to investigate possible effects of different companies and types of swine operations on the availability of N in sludge from their associated lagoons. A laboratory incubation study was conducted to quantify the availability of N (i.e. initial inorganic N plus the potentially mineralizable organic N) in the sludge. Nine sludge sources from lagoons of sow, nursery and finishing operations of three different swine companies were mixed with a loamy sand soil (200 mg total Kjeldahl N kg(-1) soil) and incubated at a water content of 0.19 g. water g(-1) dry soil and 25+/-2 degrees C for 12 weeks. Samples were taken at eight times over the 12-week period and analyzed for inorganic N (i.e. NH(4)-N and NO(3)-N) to determine mineralization of organic N in the sludge. Company and type of swine operation had no significant effects (P < 0.05) on the pattern of inorganic N accumulation over time. Thus, inorganic N accumulation from all sludge sources was fit to a first order equation [Nt = Ni + No (1-e(-kt)]. This relationship indicated that of the 200 mg of total sludge N added per kg soil, 23.5% was in the form of potentially mineralizable organic N (No) and 17.5% was in the form of inorganic N (Ni). The sum of these two pools (41%) represents an estimate of the proportion of total N in the applied sludge in plant available form after the 12 week incubation. While plant N availability coefficients were not measured in this study, the lack of significant company or type of swine operation effects on sludge N mineralization suggests that use of the same plant N availability coefficient for sludge from different types of lagoons is justifiable. The validity of this interpretation depends on the assumption that variation in other components of different sludge sources such as Cu and Zn does not differentially alter N uptake by the receiver crops.     

Potent pyrimidinetrione-based inhibitors of MMP-13 with enhanced selectivity over MMP-14

Through the use of computational modeling, a series of pyrimidinetrione-based inhibitors of MMP-13 was designed based on a lead inhibitor identified through file screening. Incorporation of a biaryl ether moiety at the C-5 position of the pyrimidinetrione ring resulted in a dramatic enhancement of MMP-13 potency. Protein crystallography revealed that this moiety binds in the S(1)(') pocket of the enzyme. Optimization of the C-4 substituent of the terminal aromatic ring led to incorporation of selectivity versus MMP-14 (MT-1 MMP). Structure activity relationships of the biaryl ether substituent are presented as is pharmacokinetic data for a compound that meets our in vitro potency and selectivity goals.     

Zn2+-dependent deoxyribozymes that form natural and unnatural RNA linkages

We report Zn(2+)-dependent deoxyribozymes that ligate RNA. The DNA enzymes were identified by in vitro selection and ligate RNA with k(obs) up to 0.5 min(-)(1) at 1 mM Zn(2+) and 23 degrees C, pH 7.9, which is substantially faster than our previously reported Mg(2+)-dependent deoxyribozymes. Each new Zn(2+)-dependent deoxyribozyme mediates the reaction of a specific nucleophile on one RNA substrate with a 2',3'-cyclic phosphate on a second RNA substrate. Some of the Zn(2+)-dependent deoxyribozymes create native 3'-5' RNA linkages (with k(obs) up to 0.02 min(-)(1)), whereas all of our previous Mg(2+)-dependent deoxyribozymes that use a 2',3'-cyclic phosphate create non-native 2'-5' RNA linkages. On this basis, Zn(2+)-dependent deoxyribozymes have promise for synthesis of native 3'-5'-linked RNA using 2',3'-cyclic phosphate RNA substrates, although these particular Zn(2+)-dependent deoxyribozymes are likely not useful for this practical application. Some of the new Zn(2+)-dependent deoxyribozymes instead create non-native 2'-5' linkages, just like their Mg(2+) counterparts. Unexpectedly, other Zn(2+)-dependent deoxyribozymes synthesize one of three unnatural linkages that are formed upon the reaction of an RNA nucleophile other than a 5'-hydroxyl group. Two of these unnatural linkages are the 3'-2' and 2'-2' linear junctions created when the 2'-hydroxyl of the 5'-terminal guanosine of one RNA substrate attacks the 2',3'-cyclic phosphate of the second RNA substrate. The third unnatural linkage is a branched RNA that results from attack of a specific internal 2'-hydroxyl of one RNA substrate at the 2',3'-cyclic phosphate. When compared with the consistent creation of 2'-5' linkages by Mg(2+)-dependent ligation, formation of this variety of RNA ligation products by Zn(2+)-dependent deoxyribozymes highlights the versatility of transition metals such as Zn(2+) for mediating nucleic acid catalysis.     

Pulmonary collectins modulate strain-specific influenza a virus infection and host responses

Collectins are secreted collagen-like lectins that bind, agglutinate, and neutralize influenza A virus (IAV) in vitro. Surfactant proteins A and D (SP-A and SP-D) are collectins expressed in the airway and alveolar epithelium and could have a role in the regulation of IAV infection in vivo. Previous studies have shown that binding of SP-D to IAV is dependent on the glycosylation of specific sites on the HA1 domain of hemagglutinin on the surface of IAV, while the binding of SP-A to the HA1 domain is dependent on the glycosylation of the carbohydrate recognition domain of SP-A. Here, using SP-A and SP-D gene-targeted mice on a common C57BL6 background, we report that viral replication and the host response as measured by weight loss, neutrophil influx into the lung, and local cytokine release are regulated by SP-D but not SP-A when the IAV is glycosylated at a specific site (N165) on the HA1 domain. SP-D does not protect against IAV infection with a strain lacking glycosylation at N165. With the exception of a small difference on day 2 after infection with X-79, we did not find any significant difference in viral load in SP-A(-/-) mice with either IAV strain, although small differences in the cytokine responses to IAV were detected in SP-A(-/-) mice. Mice deficient in both SP-A and SP-D responded to IAV similarly to mice deficient in SP-D alone. Since most strains of IAV currently circulating are glycosylated at N165, SP-D may play a role in protection from IAV infection.