A gain-of-function mutation in a cytokinin receptor triggers spontaneous root nodule organogenesis

Legume root nodules originate from differentiated cortical cells that reenter the cell cycle and form organ primordia. We show that perception of the phytohormone cytokinin is a key element in this switch. Mutation of a Lotus japonicus cytokinin receptor gene leads to spontaneous development of root nodules in the absence of rhizobia or rhizobial signal molecules. The mutant histidine kinase receptor has cytokinin-independent activity and activates an Escherichia coli two-component phosphorelay system in vivo. Mutant analysis shows that cytokinin signaling is required for cell divisions that initiate nodule development and defines an autoregulated process where cytokinin induction of nodule stem cells is controlled by shoots.     

A role for interaction of the RNA polymerase flap domain with the sigma subunit in promoter recognition

In bacteria, promoter recognition depends on the RNA polymerase sigma subunit, which combines with the catalytically proficient RNA polymerase core to form the holoenzyme. The major class of bacterial promoters is defined by two conserved elements (the -10 and -35 elements, which are 10 and 35 nucleotides upstream of the initiation point, respectively) that are contacted by sigma in the holoenzyme. We show that recognition of promoters of this class depends on the "flexible flap" domain of the RNA polymerase beta subunit. The flap interacts with conserved region 4 of sigma and triggers a conformational change that moves region 4 into the correct position for interaction with the -35 element. Because the flexible flap is evolutionarily conserved, this domain may facilitate promoter recognition by specificity factors in eukaryotes as well.     

Comparison of digital and optical hand-held refractometers for the measurement of feline urine specific gravity

Measuring urine specific gravity (USG) is an important component of urine analysis as it evaluates renal concentrating capability. The objective of this study was to quantify the difference in USG values between a hand-held optical analogue refractometer and a cat-specific digital instrument. Urine samples from 55 cats were assessed. There was a statistically significant difference between these two refractometers (P<0.001), with the optical refractometer (mean USG=1.031) consistently reading higher than the digital refractometer (mean USG=1.027). Results for a random subset of the samples (n=10) were compared with urine osmolality and both the optical and digital instruments demonstrated excellent correlation. While an accurate USG reading is important, it is unlikely that the statistical significance between the two instruments is clinically significant and, therefore, unlikely to result in a change in patient evaluation or treatment plans. While both the digital and optimal refractometers are highly correlated to the urine osmolality, making both devices valid for assessment of USG in clinical practice, this digital device is easier to read and eliminates the variability of subjective interpretation.     

Determination of the odor threshold concentrations of iodinated trihalomethanes in drinking water

Iodinated trihalomethanes (ITHMs) have been usually considered the disinfection byproducts suspected of causing medicinal odor episodes in treated water around the world. The odor threshold concentration (OTC) of mixed ITHMs (bromochloroiodo-, bromodiiodo-, chlorodiiodo-, dibromoiodo-, and dichloroiodomethane) which were previously synthesized -- because commercial standards are not available-- were determined by using two sensory techniques: flavor profile analysis (FPA), performed by an experienced panel trained in identifying odors and tastes in water; and gas chromatography coupled with olfactometry (GCO). FPA results gave a theoretical OTCs range from 0.1 to 8.9 microg/L and ITHMs were described as sweet, solvent, and medicinal products. The lowest experimental value (OTC(exp)) obtained from the six ITHMs, 0.03 microg/L, corresponded to iodoform.     

One- and two-dimensional electrophoretic identification of IgE-binding polypeptides of Lupinus albus and other legume seeds

The prevalence of food allergies in the world population requires integrated approaches to identify new potential allergens, especially those of plant origin. The aim of this work was the allergen in vitro analysis of Lupinus albus seed proteome, a promising food protein source, and the assessment of IgE cross-reactivities with other more diffused legume species. A combination of one- and two-dimensional gel electrophoresis and immunoblotting analyses with specific IgGs for band identification and lupin-sensitized patients' circulating IgEs for allergenicity studies has been used. Two lupin proteins, namely, conglutin gamma and 11S globulin basic subunits, strongly reacted with all patients' sera. Also, cross-reactivities with the homologous polypeptides of other legume species were observed. Otherwise, no reaction at all was detected with a 2S-type lupin protein. This global electrophoretic approach has allowed the identification of a new potential lupin allergen and confirmed the cross-reactivity among the legume 11S globulin basic subunits.     

Testing the MEDALUS hillslope model

In the Mediterranean environment formative erosional and runoff events are sporadic, intense and infrequent. They are difficult to study, their effects difficult to anticipate and the impact of vegetational and climatic change, whether natural or human-induced, difficult to predict. For these reasons, modelling has to augment the traditional approaches. In the Mediterranean Desertification and Land Use research programme, models have been developed to look at these problems at the hillslope (MEDALUS model) and large catchment scale (MEDRUSH model). Such models are only as good as their capacity to replicate, to an acceptable level, the magnitude, pattern in space and time and character of real world processes. These models can be tested by (a) examining their logical structure and performance against synthetic or control data (primary level) (b) evaluating their behaviour against published empirical spatial and temporal data (secondary level) and (3) by direct comparison with field test data (real world validation).The MEDALUS catena model comprises atmospheric, plant growth, overland flow and erosion, and subsurface water redistribution components involving a number of important novel features especially for dryland environments. It produces vegetation biomass and storm event runoff and sediment yield for different positions over a hillslope at different times. It is here tested against plot data for events and vegetation, soil moisture, runoff, erosion and slope armouring for annual series over several years. The model is found to perform moderately well for runoff and very well for sediment yield for event-based simulations. The results for longer simulations, tested at the secondary level, perform well in comparison with responses cited in the literature.     

Role of aldehyde oxidase in the hepatic in vitro metabolism of 3-methylindole in pigs

The metabolism of 3-hydroxy-3-methylindolenine (HMI), a recently discovered metabolite of 3-methylindole (3MI, skatole) produced by porcine liver microsomes, was investigated in vitro using porcine liver cytosol. HMI was rapidly metabolized to a single product, 3-hydroxy-3-methyloxindole (HMOI), by porcine cytosol. By the use of the selective inhibitors menadione and quinacrine, it was shown that the enzyme responsible for the oxidation of HMI into HMOI was aldehyde oxidase (AO; aldehyde:oxygen oxidoreductase, EC 1.2.3.1). The activity of AO in the conversion of HMI to HMOI was measured in a population of pigs (n = 30) with a wide range of 3MI levels in back fat (0.07-0.30 mg/kg). AO activity was found to be negatively correlated (r = -0.70; P < 0.001) with the level of 3MI in fat. The results of the present study suggest that AO plays an important role in the metabolism of 3MI in the pig and that its catalytic activity is related to an adequate 3MI clearance.     

A Novel Particle Contact Assay with the Yeast Saccharomyces cerevisiae (8 pp)

Goal, Scope and Background   Numerous xenobiotics released into surface waters are transferred to suspended particulate matter and finally attached to sediments. Aquatic organisms may be exposed to them by direct particle feeding, by physical contact with contaminated surfaces as an exposure route, and by the uptake of dissolved contaminants after equilibration via the free water phase. In order to assess potential sediment toxicity, each of these exposure routes has to be addressed. This paper presents a newly developed particle contact assay that uses the fermentation performance of a specific Saccharomyces cerevisiae strain for the assessment of toxic effects in sediments. The test procedure is based on the characteristic feature of growing yeast cells to attach to sediment particles, which are also relevant for the accumulation of contaminants. The physical contact with lipophilic contaminants mirrors an exposition pathway for the direct uptake into the cells. In order to quantitatively characterize the toxic effects of particle attached pollutants on the fermentation performance, unpolluted native reference sediment was spiked with representatives for widely distributed anthropogenic contaminants. Methods   Saccharomyces cerevisiae was established as sensitive eukaryotic microorganism for the ecotoxicological assessment of particle attached anthropogenic contaminants in freshwater sediments. For this purpose, yeast cells were cultivated in sediment samples and the resulting fermentation performance was continuously measured. Sediments artifically spiked with HCB, PCB, g-HCH, DDT, and benzo(a)pyrene and solutions of each contaminant were comparatively investigated by means of their adverse effects on yeast fermentation performance. Additionally, four native river sediments characterized by increasing levels of pollution were assessed by the yeast particle contact assay, and simultaneously by standard aquatic tests with algae, daphniae, and luminescent bacteria using pore water and elutriates. Results of the bioassays were related to specific sediment contamination with respect to metals and organic priority pollutants. Results and Discussion   In sediments spiked with PCB and benzo(a)pyrene fermentation, performance was affected extensively below concentrations inhibiting fermentation in contaminant solutions. This suggests a high efficiency of the exposure route by physical contact. The fermentation performance was only slightly affected by single lipophilic pollutants, whereas mixtures of individually spiked sediments caused critically reduced fermentation performance suggesting additive synergistic effects. Native river sediments modestly to critically polluted by hazardous organic compounds lead to a slightly to dangerously reduced fermentation performance in the yeast contact assay. These inhibitory effects were much less pronounced in the standard bioassays conducted with algae, daphniae and luminescent bacteria, applying pore waters and elutriates as sample matrices. Using pore water, inhibition was measured only in the most polluted sediment, elutriates lead to a slight inhibition of the algal growth in the undiluted sample only. These results indicate an improved sensitivity of the yeast particle contact assay compared to the standard assays, due to uptake and physical cell contact as additional routes of exposure. Conclusion   The yeast particle contact assay is a valuable tool for the assessment of ecotoxicological potential in freshwater sediments. Since the assay addresses physical contact as an exposure route, it indicates bioavailability of lipophilic compounds in sediments. Outlook   The sensitive indication of bioavailable contaminants associated to sediment particles by the newly developed yeast particle contact assay recommends it as a complementary microbial bioassay in a test battery for assessing major pathways of contaminants in whole sediments.