Kinetics of O2 uptake, leg blood flow, and muscle deoxygenation are slowed in the upper compared with lower region of the moderate-intensity exercise domain.

Six male subjects [23 yr (SD 4)] performed repetitions (6-8) of two-legged, moderate-intensity, knee-extension exercise during two separate protocols that included step transitions from 3 W to 90% estimated lactate threshold (thetaL) performed as a single step (S3) and in two equal steps (S1, 3 W to approximately 45% thetaL; S2, approximately 45% thetaL to approximately 90% thetaL). The time constants (tau) of pulmonary oxygen uptake (Vo2), leg blood flow (LBF), heart rate (HR), and muscle deoxygenation (HHb) were greater (P < 0.05) in S2 (tauVo2, approximately 52 s; tauLBF, approximately 39 s; tauHR, approximately 42 s; tauHHb, approximately 33 s) compared with S1 (tauVo2, approximately 24 s; tauLBF, approximately 21 s; tauHR, approximately 21 s; tauHHb, approximately 16 s), while the delay before an increase in HHb was reduced (P < 0.05) in S2 (approximately 14 s) compared with S1 (approximately 20 s). The Vo2 and HHb amplitudes were greater (P < 0.05) in S2 compared with S1, whereas the LBF amplitude was similar in S2 and S1. Thus the slowed Vo2 response in S2 compared with S1 is consistent with a mechanism whereby Vo2 kinetics is limited, in part, by a slowed adaptation of blood flow and/or O2 transport when exercise was initiated from a baseline of moderate-intensity exercise.     

Rhizodeposition shapes rhizosphere microbial community structure in organic soil

The aims of the study were to determine group specificity in microbial utilization of root-exudate compounds and whole rhizodeposition; quantify the proportions of carbon acquired by microbial groups from soil organic matter and rhizodeposition, respectively; and assess the importance of root-derived C as a driver of soil microbial community structure. Additions of 13C-labelled root-exudate compounds to organic soil and steady-state labelling of Lolium perenne, coupled to compound-specific isotope ratio mass spectrometry, were used to quantify group-specific microbial utilization of rhizodeposition. Microbial utilization of glucose and fumaric acid was widespread through the microbial community, but glycine was utilized by a narrower range of populations, as indicated by the enrichment of phospholipid fatty acid (PLFA) analysis fractions. In L. perenne rhizospheres, high rates of rhizodeposit utilization by microbial groups showed good correspondence with increased abundance of these groups in the rhizosphere. Although rhizodeposition was not the quantitatively dominant C source for microbes in L. perenne rhizospheres, relative utilization of this C source was an important driver of microbial group abundance in organic soil.     

A novel missense mutation in DAX-1 with an unusual presentation of X-linked adrenal hypoplasia congenita

A male presented at age 2.2 years with a 6-week history of intermittent vomiting and hyperpigmentation. Investigations showed salt wasting with hyperkalaemia, a grossly impaired cortisol response to ACTH stimulation, elevated renin and ACTH. Family history revealed that two maternal uncles had died soon after birth. A third uncle failed to thrive during infancy but improved with a course of cortisone, then being untreated until further investigation revealed adrenal insufficiency. A fourth uncle died aged 10 days, with urinary salt loss and hypoplastic adrenal glands at postmortem. Molecular studies on the proband, his mother, maternal grandmother, and surviving uncle showed a novel C to G substitution at nucleotide position 794 (missense mutation T265R) in the DAX1 (NR0B1) gene. The proband has responded well to steroid replacement but has proved sensitive to 9alpha-fludrocortisone treatment, developing hypertension on a dose of 133 microg/m(2)/day. At 8.8 years he was noted to have testicular volumes of 4 ml, despite no other evidence of secondary sexual development and prepubertal gonadotrophin levels. Novel features of this family include a novel DAX1 mutation, marked variability in age of presentation, hypertension on 'standard' doses of 9alpha-fludrocortisone and mild testicular enlargement.     

In the FVB/N HER-2/<Emphasis Type="Italic">neu</Emphasis> transgenic mouse both peripheral and central tolerance limit the immune response targeting HER-2/neu induced by <Emphasis Type="Italic">Listeria monocytogenes</Emphasis>-based vaccines

Listeria monocytogenes-based vaccines for HER-2/neu are capable of breaking tolerance in FVB/N rat HER-2/neu transgenic mice. The growth of implanted NT-2 tumors, derived from a spontaneously occurring tumor in the FVB/N HER-2/neu transgenic mouse, was significantly slower in these mice following vaccination with a series of L. monocytogenes-based vaccines for HER-2/neu. Mechanisms of T cell tolerance that exist in these transgenic mice include the absence of functional high avidity anti-HER-2/neu CD8⁺ T cells and the presence of CD4⁺CD25⁺ regulatory T cells. The in vivo depletion of these regulatory T cells resulted in the slowing in growth of tumors even without the treatment of mice with an anti-HER-2/neu vaccine. The average avidities of responsive CD8⁺ T cells to six of the nine epitopes in HER-2/neu we examined, four of which were identified in this study, are shown here to be of a lower average avidity in the transgenic mice versus wild type FVB/N mice. In contrast, the average avidity of CD8⁺ T cells to three epitopes that showed the lowest avidity in the wild-type mice did not differ between wild type and transgenic mice. This study demonstrates the ability of L. monocytogenes-based vaccines to impact upon tolerance to HER-2/neu in FVB/N HER-2/neu transgenic mice and further defines some of the aspects of tolerance in these mice.     

The genetic basis of inbreeding avoidance in house mice

Animals might be able to use highly polymorphic genetic markers to recognize very close relatives and avoid inbreeding. The major histocompatibility complex (MHC) is thought to provide such a marker because it influences individual scent in a broad range of vertebrates. However, direct evidence is very limited. In house mice (Mus musculus domesticus), the major urinary protein (MUP) gene cluster provides another highly polymorphic scent signal of genetic identity that could underlie kin recognition. We demonstrate that wild mice breeding freely in seminatural enclosures show no avoidance of mates with the same MHC genotype when genome-wide similarity is controlled. Instead, inbreeding avoidance is fully explained by a strong deficit in successful matings between mice sharing both MUP haplotypes. Single haplotype sharing is not a good guide to the identification of full sibs, and there was no evidence of behavioral imprinting on maternal MHC or MUP haplotypes. This study, the first to examine wild animals with normal variation in MHC, MUP, and genetic background, demonstrates that mice use self-referent matching of a species-specific polymorphic signal to avoid inbreeding. Recognition of close kin as unsuitable mates might be more variable across species than a generic vertebrate-wide ability to avoid inbreeding based on MHC.     

The isoepoxydon dehydrogenase gene PCR profile is useful in fungal taxonomy

This study evaluates the specificity of PCR isoepoxydon dehydrogenase (idh) primers on fungi associated with patulin production. The DNAs of 93 strains were extracted and analysed by PCR using primers of the idh gene of patulin biosynthesis. A single band at 620 bp was obtained on 17% of the analysed strains. Different molecular weight amplicons were observed in other strains. These were employed as binary characters for numerical analysis to obtain a dendrogram. Clusters were observed, which corresponded to morphological identifications in some cases. Amplicons at 400 and/or 500 bp were related to patulin non-detection for strains, whereas a 450 bp amplicon was associated with some Aspergillus and both of the Byssochlamys nivea strains tested. Hence, the idh primers are not specific for the gene and provide other amplicon products in other species. These results were useful providing (a) profiles of DNA to identify and classify fungi and (b) insights into patulin production. The DNA profiles in this study may be useful for the potential of patulin producing fungi. Obtaining multiple bands in culture-independent PCR of environmental samples by using the primers could indicate that more than one species is present.     

New methods for new questions about rhizosphere/plant root interactions

Plant and Soil - In this opinion paper we review recent methodological developments underpinning the study of roots, the rhizosphere and interactions affecting soil functions, and explore new...     

Synchrotron-Based Techniques Shed Light on Mechanisms of Plant Sensitivity and Tolerance to High Manganese in the Root Environment

Plant species differ in response to high available manganese (Mn), but the mechanisms of sensitivity and tolerance are poorly understood. In solution culture, greater than or equal to 30 µm Mn decreased the growth of soybean (Glycine max), but white lupin (Lupinus albus), narrow-leafed lupin (Lupin angustifolius), and sunflower (Helianthus annuus) grew well at 100 µm Mn. Differences in species’ tolerance to high Mn could not be explained simply by differences in root, stem, or leaf Mn status, being 8.6, 17.1, 6.8, and 9.5 mmol kg^–1 leaf fresh mass at 100 µm Mn. Furthermore, x-ray absorption near edge structure analyses identified the predominance of Mn(II), bound mostly to malate or citrate, in roots and stems of all four species. Rather, differences in tolerance were due to variations in Mn distribution and speciation within leaves. In Mn-sensitive soybean, in situ analysis of fresh leaves using x-ray fluorescence microscopy combined with x-ray absorption near edge structure showed high Mn in the veins, and manganite [Mn(III)] accumulated in necrotic lesions apparently through low Mn sequestration in vacuoles or other vesicles. In the two lupin species, most Mn accumulated in vacuoles as either soluble Mn(II) malate or citrate. In sunflower, Mn was sequestered as manganite at the base of nonglandular trichomes. Hence, tolerance to high Mn was ascribed to effective sinks for Mn in leaves, as Mn(II) within vacuoles or through oxidation of Mn(II) to Mn(III) in trichomes. These two mechanisms prevented Mn accumulation in the cytoplasm and apoplast, thereby ensuring tolerance to high Mn in the root environment.Manganese (Mn) is an essential element for plant growth, but its availability differs greatly in space and time, depending largely on the nature and amount of Mn minerals present and on the soil’s pH and redox potential. With an elaborate chemistry, Mn forms complexes with many organic and inorganic ligands. In soils, Mn has three common oxidation states, Mn(II), Mn(III), and Mn(IV), which form hydrated oxides of mixed valency; Mn is present also as numerous carbonates, silicates, sulfates, and phosphates (Lindsay, 1979). Cationic Mn²⁺ is the most common form readily absorbed by plant roots (Clarkson, 1988). The toxicity of Mn occurs in acid or waterlogged soils high in Mn minerals.Many plants have mechanisms to accommodate the large differences in Mn²⁺ in soils. At low available Mn, uptake is increased in some Poaceae by excretion of phytosiderophores of the mugineic acid family (Takahashi et al., 2003), with root phytase exudation also potentially important for acquisition of Mn when Mn availability is limited (George et al., 2014). Mechanisms in other plants include the ability of roots to decrease rhizosphere pH or excrete organic ligands (Neumann and Romheld, 2012; Lambers et al., 2015). However, the relative importance of the many complexes on Mn uptake remains unclear. Toxicity results from high Mn in leaf cell walls (Wissemeier et al., 1992; Wissemeier and Horst, 1992) and through adverse effects on symplastic proteins (Führs et al., 2008). Many plants have mechanisms that limit the adverse effects of high Mn²⁺ in soils, with numerous ligands involved in its translocation and that of other essential cations (Haydon and Cobbett, 2007). Edwards and Asher (1982) classified a range of crop and pasture species based on their ability to deal with high Mn as those that (1) limit Mn from entering the roots, (2) retain Mn in the roots, or (3) tolerate high Mn in the shoots. At the extreme are plants that hyperaccumulate more than 10,000 mg Mn kg^–1 on a dry mass (DM) basis in foliar tissues without metabolic damage (Fernando et al., 2013; van der Ent et al., 2013). Based on 15% DM of leaves, this equates to 12.1 mmol kg^–1 on a fresh mass (FM) basis. Celosia argentia, a species adapted to growth on Mn-contaminated mine tailings, accumulated more than 20,000 mg kg^–1 Mn in leaves (Liu et al., 2014). Tolerance of high Mn in shoots of some Mn hyperaccumulators has been found to occur through binding to ligands (such as malate or citrate) or sequestration in the vacuole (Fernando et al., 2010).Characteristic symptoms of Mn toxicity include chlorotic and distorted leaves with small necrotic lesions. These lesions have been shown in cowpea (Vigna unguiculata) to contain oxidized Mn and callose (Wissemeier et al., 1992), which forms as a reaction to high intracellular Ca (Kartusch, 2003). The necrotic lesions result mainly from oxidized phenolics (Wissemeier and Horst, 1992) and increased peroxidase activity in the apoplast (Horst et al., 1999). With a critical solution concentration for toxicity (10% growth reduction) of no more than 9 µm Mn, Edwards and Asher (1982) found that cotton (Gossypium hirsutum), bean (Phaseolus vulgaris), cowpea, and soybean (Glycine max) were the most sensitive species of 13 crop and pasture plants grown for 18 to 31 d at constant Mn in solution culture. By contrast, the critical concentration for sunflower (Helianthus annuus) was 7 times higher at 65 µm Mn. Sunflower was the first species found to tolerate high Mn through its sequestration in the trichomes on stems, petioles, and leaves (Blamey et al., 1986). The suspected accumulation of Mn was confirmed using wavelength dispersive x-ray spectroscopy with darkening inferred as due to insoluble higher oxides of Mn. Similarly, high Mn results in darkened trichomes of cucumber (Cucumis sativus) leaves due to oxidized Mn, as shown by the colorimetric benzidine test (Horiguchi, 1987). Watermelon (Citrullus lanatus; Elamin and Wilcox, 1986b), but not muskmelon (Citrullus melo; Elamin and Wilcox, 1986a), grown at high Mn also develops small dark spots around the leaf trichomes. Other species that sequester Mn in the trichomes include common nettle (Urtica dioica; Hughes and Williams, 1988) and Alyssum murale, a Ni hyperaccumulator (Broadhurst et al., 2009; McNear and Küpper, 2014). Thus, some plants in four families, Asteraceae, Cucurbitaceae, Urticaceae, and Brassicaceae, tolerate high Mn in shoots through Mn sequestration in or around the trichomes. The mechanisms may differ, however, because the high Mn present during development of common nettle stinging hairs decreases as plants mature (Hughes and Williams, 1988).Recently developed techniques, including those based on synchrotron radiation, allow investigations of the distribution and speciation of Mn in planta, with most research to date focused on Mn hyperaccumulators (Fernando et al., 2013). For example, Fernando et al. (2010) used x-ray absorption near-edge spectroscopy (XANES) to confirm the widely accepted view that Mn(II) predominates in seven Mn hyperaccumulators. Synchrotron-based x-ray fluorescence microspectroscopy (µ-XRF) was used by McNear and Küpper (2014) to show that the basal region of trichomes of A. murale plants grown at no more than 10 µm Mn contained Mn(II) complexed with phosphate. At 50 µm Mn in solution, however, the increased amount of Mn that had accumulated around the trichomes was present as Mn(III). Few studies, however, have used synchrotron-based techniques to investigate the mechanisms of Mn toxicity and tolerance in agronomic species despite their importance for food production in regions where soils are acidic or intermittently waterlogged. One study on cowpea, with a critical toxicity concentration of only 2 µm Mn (Edwards and Asher, 1982), has shown an accumulation of Mn-citrate in the root cap and associated mucigel within 5 min of exposure to 150 µm Mn (Kopittke et al., 2013).This study aimed to determine the distribution and speciation of Mn in fresh roots, stems, and leaves of four crop species, soybean, white lupin (Lupinus albus), narrow-leafed lupin (Lupinus angustifolius), and sunflower, which differ in tolerance to high Mn. It was hypothesized that Mn distribution and speciation would differ between Mn-sensitive soybean and the three other species. Furthermore, we considered it likely that the Mn tolerance mechanism of sunflower would differ from those of the two lupin species, which do not have darkened trichomes when grown at high Mn.     

Tight coupling of primary production and marine mammal reproduction in the Southern Ocean

Polynyas are areas of open water surrounded by sea ice and are important sources of primary production in high-latitude marine ecosystems. The magnitude of annual primary production in polynyas is controlled by the amount of exposure to solar radiation and sensitivity to changes in sea-ice extent. The degree of coupling between primary production and production by upper trophic-level consumers in these environments is not well understood, which prevents reliable predictions about population trajectories for species at higher trophic levels under potential future climate scenarios. In this study, we find a strong, positive relationship between annual primary production in an Antarctic polynya and pup production by ice-dependent Weddell seals. The timing of the relationship suggests reproductive effort increases to take advantage of high primary production occurring in the months after the birth pulse. Though the proximate causal mechanism is unknown, our results indicate tight coupling between organisms at disparate trophic levels on a short timescale, deepen our understanding of marine ecosystem processes, and raise interesting questions about why such coupling exists and what implications it has for understanding high-latitude ecosystems.