High resolution proton NMR spectroscopy of human amniotic fluid

Human amniotic fluid (HAF) is a dynamic system whose characteristics depend on continuous interchanges between fetal and maternal circulations. HAF reflects not only the environment of the fetus but may also provide information about fetal development or pathology. The concentration of HAF constituents varies with gestational age and pathological states. The number of the compounds currently implicated in fetal developmental pathology are relatively few. Currently used assay methods are not adequate to totally explain or predict the complex biochemistry of the fetus. The purpose of this work was to investigate HAF with NMR spectroscopy. In the present study HAF was obtained from 47 women undergoing routine amniocentesis. Cells were separated for karyological analysis and the supernatant was acid-extracted, lyophilized and re-suspended in D₂0 resulting in a concentration increase over native fluid. ¹H NMR spectra were obtained at 360 MHz and 60 MHz. Eighteen compounds including several amino acids, were identified using parallel reference and standard addition protocols. NMR spectroscopy detected compounds of known clinical importance including glucose, leucine, isoleucine, lactate and creatinine. In conclusion, we have demonstrated that a number of physiologically relevant compounds are readily observable in HAF using ¹H NMR spectroscopy. This technique can currently provide valuable information regarding HAF composition and has the potential of being used in vivo in the future.     

Assessing the utility of dissolved organic matter photoreactivity as a predictor of in situ methylmercury concentration

Methylmercury(MeHg) bioaccumulation is a growing concern in ecosystems worldwide. The absorption of solar radiation by dissolved organic matter(DOM) and other photoreactive ligands can convert MeHg into less toxic forms of mercury through photodemethylation. In this study, spectral changes and photoreactivity of DOM were measured to assess the potential to control photoreactions and predict in situ MeHg concentration. Water samples collected from a series of lakes in southwestern Nova Scotia in June, August, and September were exposed to controlled ultraviolet-A(UV-A) radiation for up to 24 hr. Dissolved organic matter photoreactivity, measured as the loss of absorbance at 350 nm at constant UV-A irradiation, was positively dependent on the initial DOM concentration in lake waters(r~2=0.94). This relationship was consistent over time with both DOM concentration and photoreactivity increasing from summer into fall across lakes. Lake in situ MeHg concentration was positively correlated with DOM concentration and likely catchment transport in June(r = 0.77) but not the other sampling months. Despite a consistent seasonal variation in both DOM and Fe, and their respective correlations with MeHg, no discernable seasonal trend in MeHg was observed. However, a 3-year dataset from the 6 study lakes revealed a positive correlation between DOM concentration and both Fe(r = 0.91) and MeHg concentrations(r = 0.51) suggesting a more dominant landscape mobility control on MeHg.The DOM-MeHg relationships observed in these lakes highlights the need to examine DOM photoreactivity controls on MeHg transport and availability in natural waters particularly given future climate perturbations.     

Spatial variation in dimethylsulfoniopropionate (DMSP) production in <Emphasis Type="Italic">Ulva lactuca</Emphasis> (Chlorophyta) from the Northeast Pacific

Although dimethylsulfoniopropionate (DMSP) has a variety of functions in marine macroalgae including that of a cryoprotectant, an osmolyte, a way to remove excess sulfur and energy, an antioxidant, and an allelopathic precursor, the latter two functions are believed to be the most important in Ulva lactuca L. (=U. fenestrata) in intertidal populations on the coast of Washington state, USA. The present study found significant variation in DMSP concentrations among U. lactuca collected in May 2005 from six sites ranging from 47°54.45′N (Possession Point, Whidbey Island, WA, USA) to 48°30.55′N (Shannon Point Beach, Anacortes, WA, USA), and also among individuals within sites, and among tissues (basal tissues near the holdfast, middle of the blades, and tips). Concentrations ranged from 37 to 224 μmol g⁻¹ fresh mass (FM). In several 10-day experiments between July 2001 and August 2004 with U. lactuca collected from several places on the coast of Washington, the effects of nutrient level (DIN), light intensity and wavelength, and grazing by the herbivorous gastropod Lacuna vincta, were examined. None of these manipulations resulted in DMSP concentrations that differed significantly from controls, and variance in DMSP concentrations within each experiment was very low. Although DMSP concentrations in U. lactuca may be affected by factors not tested in these experiments, it is also possible that the observed spatial differences reflect constitutive genotypic or phenotypic differences among geographically separated U. lactuca populations or among cryptic Ulva species. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Determinants of male spacing behavior in microtines: an experimental manipulation of female spatial distribution and density

Theory on microtine mating systems predicts that male spacing behavior will be related to female spatial and temporal distribution. However, data from a natural population of field voles, Microtus agrestis, indicated a potential influence of female density on the spacing behavior of males. Therefore, I experimentally investigated the relative importance of female density and female spatial distribution for the spacing behavior of males in M. agrestis. Males were radio-tracked in enclosed natural habitats in which females at different densities were placed in two different spatial arrangements: clumped versus even distribution. Female density was the main factor determining male spacing behavior. At the high female density males had smaller home ranges and moved shorter distances between radio-tracking recordings. Also, home ranges were more exclusive at high female density. However, since there was a significant positive correlation between home range size and range overlap, range exclusiveness seemed to be influenced by female density indirectly through the effect of range size. Female spatial distribution, on the other hand, had no influence on male home range exclusiveness.     

Song overproduction and selective attrition lead to song sharing in the field sparrow (Spizella pusilla)

Summary Male field sparrows settling on particular territories for the first time often sing two or more song types. Vocal experience with neighboring males then leads in most cases to the deletion from the male's repertoires of all but one song type. In 2 years, settling males retained the song type that most closely resembled that of a neighbor. Focal observations in 1 year revealed that the most actively singing neighbor was the one the new birds resembled. Songs were not selected on the basis of their absolute acoustic characteristics, but rather by their resemblance to neighbors' songs. After the attrition process is complete, males retain their song for the rest of their lives. Evidence supports the hypothesis that the actual acquisition of a song need not take place at the time and place as males occupy territories. Rather, males may acquire an abundance of song models earlier in life and then select songs from this repertoire that most closely resemble those of their neighbors. The development of mature repertoires and of local dialects is the outcome of one process in this species.     

Morphogenesis of opal teeth in calanoid copepods

Opal teeth of calanoid copepods develop early in the premolt phase of the molt cycle. They form in the apolysis space beneath the old tooth row on the mandibular gnathobase. We examined stages of tooth formation inNeocalanus spp. andCalanus pacificus. Apolysis occurs early in the distal gnathobase, then several epidermal cell types participate sequentially in formation of new teeth. Fibrous molds in the shapes of the new teeth are extruded onto the epidermal surface, then additional materials, probably proteinaceous, are secreted into them. Tooth molds next are linked to a gland in the proximal part of the gnathobase by ducts of an unusual type, lamellar-walled ducts. Silicification follows, apparently using highly osmiophilic material supplied by the proximal gland. Opal is laid down at the outer periphery of the mold then thickens toward the attachment of the mold to newly formed chitin at its base. During apolysis the epidermal cells move proximally without breaking ducts that connect small dermal glands with pores in the sides of the opal teeth. The molds for the new teeth form surrounding these ducts, and new pores result. The glands are like the ciliary exocrine glands seen in other Crustacea, with the duct deriving from fusion of the axonemes of a cilium. Presumably the glands secrete a substance into food newly broken by the teeth. This could be a toxin or a digestive enzyme.     

The toxicity of heavy metals to embryos of the American oyster Crassostrea virginica

The acute toxicity of 11 heavy metals to embryos of the American oyster Crassostrea virginica was studied and the concentrations at which 50% of the embryos did not develop were determined. The most toxic metals and their LC₅₀ values were mercury (0.0056 ppm), silver (0.0058 ppm), copper (0.103 ppm) and zinc (0.31 ppm). Those metals that were not as toxic and their LC₅₀ values were nickel (1.18 ppm), lead (2.45 ppm) and cadmium (3.80 ppm). Those metals that were relatively non-toxic and their LC₅₀ values were arsenic (7.5 ppm), chromium (10.3 ppm) and manganese (16.0 ppm). Aluminum was non-toxic at 7.5 ppm, the highest concentration tested.