Nitrogenase activity and respiration of cultures of Rhizobium spp. with special reference to concentrations of dissolved oxygen

Studies of nitrogenase in cultures of the cowpea rhizobia (Rhizobium spp.) strains 32H1 and CB756 are reported. Preliminary experiments established that, even when agar cultures were grown in air, suspensions of bacteria prepared anaerobically from them were most active at low concentrations of free dissolved O2. Consequently, assays for activity used low concentrations of O2, stabilized by adding the nodule pigment leghaemoglobin. In continuous, glutamine-limited cultures of 32H1, nitrogenase activity appeared only when the concentration of dissolved O2 in the cultures approached 1 muM. Lowering the glutamine concentration in the medium supplied to the culture from 2 to 1 mM halved the cell yield and nitrogenase activity was also diminished. Omitting succinate from the medium caused the concentration of dissolved O2 to rise and nitrogenase activity was lost. Upon restoration of the succinate supply, the O2 concentration immediately fell and nitrogenase was restored. The activity doubled in about 8 h, whereas the doubling time of this culture was 14 h. Sonic extracts of 32H1 cells from continuous cultures with active nitrogenase contained components reacting with antiserum against nitrogenase Mo-Fe protein from soybean bacteroids. Continuous cultures grown at higher O2 concentration, with only a trace of active nitrogenase, contained less of these antigens and they were not detected in highly aerobic cultures. Nitrogenase activity of a continuous culture was repressed by NH+4; the apparent half-life was about 90 min. Cells of 32H1 from a continuous culture growing at between 30 and 100 muM dissolved O2 possessed a protective mechanism which permitted respiration to increase following exposure to a rapid increase in O2 concentration from low levels (O2 shock). This effect disappeared as the O2 concentration for growth was reduced towards 1 muM.     

Gated blood pool imaging following 99mTc stannous pyrophosphate imaging

Cardiac patients who have undergone 99mTc-stannous pyrophosphate (99mTc-PYP) myocardial imaging can be injected 24 hours later with 99mTc-pertechnetate (99mTc04) to assess left ventricular function. Reduction of 99mTc04 by tin remaining in the blood following the stannous pyrophosphate injection causes labeling of the red cells by 99mTc04 and the creation of a vascular tracer suitable for electrocardiographically gated imaging.     

Ruminal degradation of tannin‐treated legume meals

The inefficiency of protein utilisation by ruminants fed protein concentrates (based on legume meals) causes serious economic loss and environmental damage owing to their rapid hydrolysis and deamination in the rumen. Thus efforts aimed at slowing the ruminal fermentation of such feeds are needed, and recent studies have observed potentially positive effects of tannins on ruminant nutrition under certain circumstances. Tannins are a complex group of naturally occurring plant polyphenols characterised by their ability to bind with proteins. This property of tannins is considered responsible for the decreased ruminal digestibility of forages both in vivo and in vitro. Under that perspective, commercial tannic acid was added at three proportions (10, 25 and 50 g kg^?1 on a dry matter basis) to four different legume meals (horse bean, kidney bean, soybean and pea), and the effect on in situ dry matter and crude protein ruminal disappearance was assessed. The results confirmed the dose‐dependent (although not persistent after 48 h) slowing of in situ digestibility, this effect being significant at the highest tannin treatment when compared with untreated samples. Scanning electron microscopy revealed that soybean seed endosperm cell walls were protected from digestion by the ruminal microbiota, while the digestion of starch granules was relatively unaffected by tannic acid. Electrophoresis of the protein fractions confirmed the lower digestibility of tannin‐treated seeds as well as the relative lack of alteration of the electrophoretic profile of individual proteins. Implications for the digestion of concentrates in ruminants are discussed. Copyright © 2004 Society of Chemical Industry     

Bioassay for determining 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEs) in human hepatoma Hepg2 cells

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), and benz[a]anthracene (BA) highly induce cytochrome P4501A1, determined by aryl hydrocarbon hydroxylase (AHH) activity, in human hepatoma HepG2 cells within 24 h. AHH activity induced by TCDD and TCDF persists for at least 48 h. In contrast, AHH activity induced by BA rapidly declines, although the amounts applied are 4-5 orders of magnitude higher than those of TCDD or TCDF. AHH induction in HepG2 cells differs from that in rat hepatoma cells H4IIEC3/T in two aspects: (1) HepG2 cells are 20 times less sensitive to the test compounds than H4IIEC3/T cells. (2) TCDF-induced AHH activity does not persist in the rat cells. The results suggest that human HepG2 cells, because of their low sensitivity, are inferior to rat H4IIEC3/T cells for determining TCDD equivalents in environmental samples. They may be useful for investigating species dependent differences in the toxicokinetics of individual polyhalogenated aromatic hydrocarbon congeners.     

Purification, characterization, and spasmogenic activity of neurotensin from the toad Bufo marinus

Neurotensin (NT) was isolated from an extract of the intestine of the cane toad, Bufo marinus and its primary structure established as: pGlu-Ala-Ile-Val-Ser-Lys-Ala-Arg-Arg-Pro-Tyr-Ile-Leu. This amino acid sequence shows five substitutions (Leu2 --> Ala, Tyr3 --> Ile, Glu4 --> Val, Asn5 --> Ser, and Pro7 --> Ala) compared with bovine NT. Synthetic Bufo NT (pD2 = 8.05 +/- 0.28) was equipotent and equally effective as bovine NT (pD2 = 8.24 +/- 0.38) in producing spasmogenic contraction of isolated segments of toad small intestine. However, the maximum response produced by Bufo NT was only 35 +/- 2% of that produced by substance P. The potencies, but not the maximum responses, to Bufo and bovine NT were significantly (p < 0.05) attenuated by pre-treatment with atropine but neither parameter was significantly diminished by tetrodotoxin and indomethacin. The data suggest that the action of NT involves interaction with receptors on toad intestinal smooth muscle that recognize the C-terminal region of NT (residues 8-13) that has been fully conserved during evolution of tetrapods. Contractile activity is mediated, at least in part, by release of acetylcholine.     

Molecular modeling of immunoglobulin light chains implicates hydrophobic residues in non-amyloid light chain deposition disease

Light chain deposition disease is a severe complication of certain immunoproliferative disorders, due to the secretion of a monoclonal light chain which precipitates close to basement membranes of several tissues. A kappa isotype restriction and an unusual frequency of a variable region subgroup (VkappaIV) suggest that precise structural features govern the propensity of pathogenic light chains to precipitate in extracellular spaces. We studied primary structures of light chains from six patients with light chain deposition disease in comparison with light chains from other pathological conditions. Sequence alignment revealed the presence of certain amino acids only in light chain deposition disease, in particular non-polar replacing hydrophilic residues. To determine the role of these residues, structures of the variable domain from four kappa chains belonging to VkappaI and VkappaIV subgroups responsible for deposition disease were modeled using known immunoglobulins as templates. The most evident structural features shared by all pathogenic light chains were hydrophobic residues exposed to the solvent in complementarity determining regions 1 or 3. In contrast to immunoglobulin light chain- related amyloidosis, where deposition of organized material might be due to electrostatic interactions between light chain dimers, hydrophobic interactions could enhance amorphous precipitation in non- amyloid light chain deposition disease.      

Study on Composition Distribution and Ferromagnetism of Monodisperse FePt Nanoparticles

Monodisperse FePt nanoparticles with size of 4.5 and 6.0 nm were prepared by simultaneous reduction of platinum acetylacetonate and thermal decomposition of iron pentacarbonyl in benzylether. The crystallography structure, size, and composition of the FePt nanoparticles were examined by X-ray diffraction and transmission electron microscopy. Energy dispersive X-ray spectrometry measurements of individual particles indicate a broad compositional distribution in both the 4.5 and 6 nm FePt nanoparticles. The effects of compositional distribution on the phase-transition and magnetic properties of the FePt nanoparticles were investigated.     

Effect of oil composition on both adjuvant-induced arthritis and delayed hypersensitivity to purified protein derivative and peptidoglycans in various rat strains

Various risk factors were evaluated to explain a significantly greater incidence of coronary heart disease in men of Japanese ancestry resident in Hawaii compared with men resident in Japan. The independent predictors of incidence of coronary heart disease in both Japan and Hawaii were systolic blood pressure, serum cholesterol, relative weight and age. These factors appeared to influence incidence similarly in both areas because in each case the correlation coefficients for Japan and Hawaii did not differ significantly. The hypothesis that the greater incidence in Hawaii could be attributed to differences in levels of these risk factors was tested with the Walker-Duncan method. The four variable multiple logistic function describing the probability of coronary heart disease in Japan was applied to the cohort characteristics observed in Hawaii. The estimated incidence thus obtained was not significantly different from that actually observed in the men resident in Hawaii. Therefore the increased coronary risk profile in Hawaii compared with Japan can account for the greater incidence of coronary heart disease in the former. Current cigarette smoking was significantly related to the risk of coronary heart disease in Hawaii but not in Japan. This difference requires further investigation.     

Esophageal perforation. An unusual presentation with a benign clinical course

A case of esophageal perforation occurring in a 75-year-old man is presented. The clinical presentation was benign and the diagnosis of esophageal perforation was made incidental to an upper gastrointestinal barium examination. Conservative management consisting of nasogastric suctioning, antibiotics and antacids was effective in minimizing the clinical course. This represents a rare instance of a successful nonsurgical approach to esophageal perforation.