Experimental confirmation of calculated phases and electron density profile for wet native collagen.

An experimental procedure is developed to phase the reflections obtained in x-ray diffraction investigations of collagen in native wet tendons. Phosphotungstic acid was used for isomorphous addition in phase determination and was located by electron microscopy. Structure factors (with phases) were obtained from the electron microscopy data for the heavy metal. Structure-factor magnitudes for collagen with and without the heavy metal were obtained from the x-ray diffraction data. The first 10 orders were investigated. Standard Argand diagrams provided two solutions for each of these, except the weak sixth order. In each case, one of the two possible solutions agrees well with the phases proposed on theoretical grounds by Hulmes et al. The present results suggest that their other proposed phases are probably correct. An electron density profile along the unit cell of the fibril is presented that shows a distinct step, as expected on the basis of the hole-overlap model. The overlap region is 48% of the length of the unit cell.     

Isolation of bacteriophage 14-lysogenized Salmonella from the freshwater aquarium snail Apullaria.

A naturally occurring Salmonella mikawasima serologically converted by phage 14 (6,7,14:y:e,n,z15) has been isolated for the first time. An S. tennessee variant seroconverted by phage 14 (6,7,14:z29:-) was also isolated. The source of these salmonellae was the common freshwater aquarium snail Ampullaria. Phage 14 prepared from these serovariants was lytic for S. bovis-morbificans (6,8:r:1,5) and for S. hadar (6,8:z10:e,n,x).     

Hysteretic nature of NADH substrate inhibition of bovine liver glutamate dehydrogenase.

NADH substrate inhibition of bovine liver glutamate dehydrogenase appears to be eliminated at enzyme concentrations above 0.5mg/ml. Since the inhibition cannot be restored by preincubation of the enzyme with any substrate or product combination, the release of inhibition had previously been considered the result of enzyme polymerization. Benzene-saturated solutions, however, increase the extent of enzyme polymerization without affecting the NADH inhibition. These and related control measurements demonstrate that the release of substrate inhibition is the result of a hysteretic transition of an enzyme central and transitory complex.     

Isolation of salmonellae and other potential pathogens from the freshwater aquarium snail Ampullaria.

The freshwater aquarium snail (Ampullaria spp.) was demonstrated to carry as many as 10(8) viable mesophilic bacteria per g of meat plus shell. Some 16 genera of bacteria were identified, with gram negatives predominating. Enrichment culture techniques enabled the isolation of salmonellae from 24 to 42 lots of 200 g each. The salmonellae comprised eight different serotypes, including Salmonella newport, Salmonella saint-paul, and Salmonella infantis. This association of salmonellae with snails may contribute to cases of human salmonellosis, since other aquarium species have already been shown to contribute to many such cases. The snails were also found to commonly harbor Pseudomonas aeruginosa and, occasionally, Edwardsiella tarda.     

Phosphate compounds in rat erythrocytes and reticulocytes.

The concentrations of glycolytic intermediates, including 2,3-diphosphoglycerate, were similar in rat reticulocytes and erythrocytes. There were striking differences, however, in the content and kind of water-soluble nucleotides. Reticulocytes contained much higher concentrations of ATP, GTP, UTP and CTP and had nucleotides not detected in the mature cell including UDP-acetylhexosamine, guanosine diphosphomannose and an unidentified cytidine compound. A large fraction of the total GTP found in the reticulocyte was in the form of a 1:1 complex of ferric iron with GTP.     

Electron addition to the (FeO2) unit of oxyhaemoglobin Glycera

Exposure of glassy solutions containing the monomeric fraction of the oxyhaemoglobin of the polychaete annelid Glycera dibranchiata to 60Co gamma-rays at 77 K resulted in electron addition to the (FeO2) moiety. The form of the g tensor components obtained from the ESR spectrum indicates that the spin-density on oxygen is much greater than that observed for similar paramagnetic centres formed in haemoglobin A or myoglobin. A major difference between these monomer haem units and normal haem units is that the distal histidine (E7 58) is replaced by leucine. We therefore postulate that the oxygen in the (FeO2)- units formed in haemoglobin A and myoglobin is hydrogen-bonded to the NH group of the distal histidine, whilst that of the (FeO2)- units in haemoglobin Glycera are not hydrogen-bonded. However, on annealing to approx. 160 K the spectrum changed irreversibly into one resembling those for (FeO2)- units in haemoglobin A and myoglobin. We postulate that this is caused by hydrogen-bonding to a water molecule in the haem pocket. Exposure of the polymeric fractions of haemoglobin Glycera to gamma-rays gave an (FeO2)- unit with an ESR spectrum remarkably similar to that obtained from oxymyoglobin. The X-ray structure of this protein is unknown but we suggest that our results could indicate the presence of a distal histidine in this material.     

Microbial communities from Arctic marine sediments respond slowly to methane addition during ex situ enrichments

Anaerobic methanotrophic archaea (ANME) consume methane in marine sediments, limiting its release to the water column, but their responses to changes in methane and sulfate supplies remain poorly constrained. To address how methane exposure may affect microbial communities and methane- and sulfur-cycling gene abundances in Arctic marine sediments, we collected sediments from offshore Svalbard that represent geochemical horizons where anaerobic methanotrophy is expected to be active, previously active, and long-inactive based on reaction-transport biogeochemical modelling of porewater sulfate profiles. Sediment slurries were incubated at in situ temperature and pressure with different added methane concentrations. Sediments from an active area of seepage began to reduce sulfate in a methane-dependent manner within months, preceding increased relative abundances of anaerobic methanotrophs ANME-1 within communities. In previously active and long-inactive sediments, sulfur-cycling Deltaproteobacteria became more dominant after 30 days, though these communities showed no evidence of methanotrophy after nearly 8 months of enrichment. Overall, enrichment conditions, but not methane, broadly altered microbial community structure across different enrichment times and sediment types. These results suggest that active ANME populations may require years to develop, and consequently microbial community composition may affect methanotrophic responses to potential large-scale seafloor methane releases in ways that provide insight for future modelling studies.