Conformationally constrained opioid peptide analogs with novel activity profiles

Novel conformationally constrained opioid peptide analogs with antagonist, mixed agonist/ antagonist or agonist properties were developed. TIP(P)-related antagonists showed unprecedented antagonist potency and receptor selectivity, and may have potential for use in analgesia in combination with agonists. A definitive model of their receptor-bound conformation was developed. Three prototype mixed agonist/ antagonists were discovered. They represent the only known compounds with this pharmacological profile and, as expected, one of them was shown to be a potent analgesic and to produce no dependence and less tolerance than morphine. Novel dipeptide derivatives turned out to be potent and selective agonists. Because of their low molecular weight and lipophilic character, these compounds may cross the blood-brain barrier and, thus, may have potential as centrally acting analgesics.     

Photosynthetic pathways and the ecological distribution of the chenopodiaceae in Isreal

Summary Fifty-four species of the Chenopodiaceae in Israel were examined for their anatomical features, ¹³C values, habitat and phytogeographical distribution. 17 species have ¹³C values between -20 and -30and non-Kranz anatomy (NK) and are therefore considered as C₃ plants. 37 species have ¹³C values between -10 and -18 and Kranz or C₄-Suaeda type anatomy and are therefore considered as C₄ plants. Some C₄ plants have leaf structure which seems to be intermediate between the Kranz and the C₄-Suaeda type of leaf anatomy.The segregation of the species into photosynthetic groups shows tribal and phytogeographical grouping. Most of the C₃ Chenopods are either mesoruderal plants or coastal halophytes, with a distribution area which covers the Euro-Siberian as well as the Mediterranean phytogeographical regions. The C₄ Chenopods are mainly desert or steppe xerohalophytes with a distribution area which includes the Saharo-Arabian and/or Irano-Turanian phytogeographical regions.     

Spatial and temporal variation in carbon isotope discrimination in prairie graminoids

We present the results of a 5-year examination of variation in the stable carbon isotope composition () of three C₃ graminoid species from a Sandhills prairie: Agropyron smithii, Carex heliophila and Stipa comata. Although consistent species-specific patterns for mean were seen, there was also significant and substantial among-year and within-season variation in . A smaller contribution to variation in came from topographic variation among sampling sites. Effects of species, year, season and topography contribute to variation in in an additive manner. An association between climate and exists that is consistent with previous work suggesting that reflects the interplay between photosynthetic gas exchange and plant water relations. Within the growing season, the time over which integrates plant response to the environment ranges from days to months.     

Stimulation of crystallin synthesis in embryonic brain cells in culture

Summary Expression of -crystallin, a lens-specific protein, in 6-day-old chick embryonic brain cells was examined in situ and in vitro. The presence of minute amounts of -crystallin and its mRNA (-mRNA) in brain cells in situ was demonstrated by immunoblot and Northern blot analysis. In spreading cultures of the brain cells, -crystallin and -mRNA showed a significant increase from their in situ level. Immunohistological staining (peroxidase antiperoxidase) with monospecific anti-serum against -crystallin revealed that -producers were both epithelial cells and dendritic cells. Neither lentoidogenesis nor -crystallin expression was observed. Stimulation of -crystallin synthesis in cultured brain cells differed when compared with transdifferentiating cultures of neural retina cells. In the latter, -crystallin synthesis occurred concomitantly with differentiation of morphologically distinct lens cells containing -crystallin.     

Modifications by chronic intermittent hypoxia and drug treatment on skeletal muscle metabolism

The energy metabolism was evaluated in gastrocnemius muscle from 3-month-old rats subjected to either mild or severe 4-week intermittent normobaric hypoxia. Furthermore, 4-week treatment with CNS-acting drugs, namely, -adrenergic (-yohimbine), vasodilator (papaverine, pinacidil), or oxygen-increasing (almitrine) agents was performed. The muscular concentration of the following metabolites was evaluated: glycogen, glucose, glucose 6-phosphate, pyruvate, lactate, lactateto-pyruvate ratio; citrate, -ketoglutarate, succinate, malate; aspartate, glutamate, alanine; ammonia; ATP, ADP, AMP, creatine phosphate. Furthermore the Vmax of the following muscular enzymes was evaluated: hexokinase, phosphofructokinase, pyruvate kinase, lactate dehydrogenase; citrate synthase, malate dehydrogenase; total NADH cytochrome c reductase; cytochrome oxidase. The adaptation to chronic intermittent normobaric mild or severe hypoxia induced alterations of the components in the anaerobic glycolytic pathway [as supported by the increased activity of lactate dehydrogenase and/or hexokinase, resulting in the decreased glycolytic substrate concentration consistent with the increased lactate production and lactate-to-pyruvate ratio] and in the mitochondrial mechanism [as supported by the decreased activity of malate dehydrogenase and/or citrate synthase resulting in the decreased concentration of some key components in the tricarboxylic acid cycle]. The effect of the concomitant pharmacological treatment suggests that the action of CNS-acting drugs could be also related to their direct influence on the muscular biochemical mechanisms linked to energy transduction.     

Intracellular sites of the synthesis of sweet potato mitochondrial F1ATPase subunits

Summary Five subunits (-, -, -, - and -subunits) of the six -and -subunits) in the F₁ portion (F₁ATPase) of sweet potato (Ipomoea batatas) mitochondrial adenosine triphosphatase were isolated by an electrophoretic method. The - and -subunits were not distinguishable immunologically but showed completely different tryptic peptide maps, indicating that they were different molecular species. In vitro protein synthesis with isolated sweet potato root mitochondria produced only the -subunit when analyzed with anti-sweet potato F₁ATPase antibody reacting with all the subunits except the -subunit. Sweet potato root poly(A)⁺RNA directed the synthesis of six polypeptides which were immunoprecipitated by the antibody: two of them immunologically related to the -subunit and the others to the - and -subunits. We conclude that the -subunit of the F₁ATPase is synthesized only in the mitochondria and the -, - and -subunits are in the cytoplasm.     

Hydrogen isotope discrimination in higher plants: Correlations with photosynthetic pathway and environment

Summary The ratio of deuterium to hydrogen (expressed as D) in hydrogen released as water during the combustion of dried plant material was examined. The D value (metabolic hydrogen) determined on plant materials grown under controlled conditions is correlated with pathways of photosynthetic carbon metabolism. C₃ plants show mean D values of-132 for shoots and -117 for roots; C₄ plants show mean D values of -91 for shoots and-77 for roots and CAM plants a D value of-75 for roots and shoots. The difference between the D value of shoot material from C₃ and C₄ plants was confirmed in species growing under a range of glasshouse conditions. This difference in D value between C₃ and C₄ species does not appear to be due to differences in the D value (tissue water) in the plants as a result of physical fractionation of hydrogen isotopes during transpiration. In C₃ and C₄ plants the hydrogen isotope discrimination is in the same direction as the carbon isotope discrimination and factors contributing to the difference in D values are discussed. In CAM plants grown in the laboratory or collected from the field D values range from-75 to +50 and are correlated with ¹³C values. When deprived of water, the D value (metabolic hydrogen) in both soluble and insoluble material in leaves of Kalanchoe daigremontiana Hamet et Perr., becomes less negative. These changes may reflect the deuterium enrichment of tissue water during transpiration, or in field conditions, may reflect the different D value of available water in areas of increasing aridity. Whatever the origin of the variable D value in CAM plants, this parameter may be a useful index of the water relations of these plants under natural conditions.     

Effect of organic and inorganic fertigation on yields, δ15N values, and δ13C values of tomato (Lycopersicon esculentum Mill. cv. Saturn)

We examined the effects of fertilizer application, especially the effects of fertigation and types of fertilizer (inorganic and organic) on yields and ¹⁵N and ¹³C values of tomato (Lycopersicon esculentum Mill. cv. Saturn). Fertigation is a method in which an appropriate diluted liquid fertilizer is applied to the plants each time they are drip-irrigated. We developed a method of organic fertigation using corn steep liquor (CSL) as the liquid fertilizer, because it is an industrial byproduct of cornstarch manufacture and can be used very effectively. We compared fruit yield, mineral content, ¹⁵N value, and ¹³C value of tomatoes grown under three different fertilizer treatments, basal dressing: basal dressing with granular chemical fertilizer; inorganic fertigation: fertigation with liquid chemical fertilizer; and organic fertigation: fertigaion with CSL. Mineral contents of tomatoes grown with basal dressing were generally lower than those grown under either fertigation treatment. These results indicated that yields and mineral contents were influenced more by the method of fertilizer application than by whether the fertilizers were inorganic or organic. There were, however, significant differences in the ¹⁵N values of tomato fruits grown under different types of fertilizer applications, especially between inorganic and organic fertilizers. The ¹⁵N value of the chemical fertilizer used for basal dressing was 0.81 ± 0.45{}, that of the chemical fertilizer for fertigation was 0.00 ± 0.04{}, and that of CSL was 8.50 ± 0.71{}. The ¹⁵N values of the soils reflected the ¹⁵N values of the fertilizers. Moreover, the ¹⁵N values of the fruits corresponded to the ¹⁵N values of the applied fertilizers. The ¹⁵N values were 3.18 ± 1.34{} in the fruits grown with a basal dressing of chemical fertilizer, 0.30 ± 0.61 in those grown under inorganic fertigation, and 7.09 ± 0.68 in those grown under organic fertigation. On the other hand, although the ¹³C values in the soil also reflected the ¹³C values of the applied fertilizers, there was no significant difference in the ¹³C values of fruits among the different treatments. In conclusion, because the ¹⁵N values of fertilizers correlated well with those of the fruits, it may be possible to use ¹⁵N values as an indicator of organic products.     

Using stable hydrogen and oxygen isotope measurements of feathers to infer geographical origins of migrating European birds

Successful application of stable-hydrogen isotope measurements (D_f) of feathers to track origins of migratory birds and other wildlife requires a fundamental understanding of the correlation between D_f and deuterium patterns in rainfall (D_p) over continental scales. A strong correlation between D_p and D_f has been confirmed for birds and insects in North America, but not yet for other continents. Here, we compare D_f data from resident European birds to new D_p basemaps for Europe. Three maps, representing growing-season and mean annual D_p estimates from an elevation-explicit, detrended interpolation model and growing-season D_p estimates from simple Kriging, all indicate that strong isotope gradients occur across Europe with a general depletion occurring in a northeast direction. The feather data, representing 141 individuals of 25 avian species from 38 sites, ranged from –131 to –38. Regression analysis showed that strong correlations existed between both mean annual and growing-season D_p estimated by detrended interpolation and D_f of non-aquatic and non-corvid birds (r²=0.66 and 0.65, respectively). We also examined mean annual and growing-season ¹⁸O_p vs. ¹⁸O_f for our samples. Both oxygen regressions were similar (r²=0.56 and 0.57, respectively) but poorer than for deuterium. Our study reveals that D measurements of feathers from migratory birds in Europe may be used to track their origin and movements, and so provide a powerful investigative tool for avian migration research in Europe.     

Multiple Phosphorylation Sites and Quaternary Organization of Guanine-Nucleotide Exchange Complex of Elongation Factor-1 (EF-1βγδ/ValRS) Control the Various Functions of EF-1α

The eukaryotic guanine-nucleotide exchange factor commonly called elongation factor-1 (EF-1), comprises four different subunits including valyl-tRNA synthetase (EF-1/ValRS). The factor is multiply-phosphorylated by three different protein kinases, protein kinase C, casein kinase II and cyclin dependent kinase 1 (CDK1). EF-1/ValRS is organized as a macromolecular complex for which we propose a new structural model. Evidence that EF-1/ValRS is a sophisticated supramolecular complex containing many phosphorylation sites, makes it a potential regulator of any of the functions of its partner EF-1, not only involved in protein synthesis elongation, but also in many other cellular functions.     

The role of vanadium in green plants

In a series of experiments, it is demonstrated that the trace element vanadium (4·10^-7 g-at/l as NH₄VO₃) has a considerable positive influence on the synthesis of -aminolevulinic acid (-ALA) in the autotrophically growing green algaChlorella pyrenoidosa, the effect being visible by an enhanced output of the amino acid into the culture medium in presence of levulinic acid (LA). The level of intracellularly accumulated -ALA, however, is not changed in presence of the metal. The V-effect on exogenous found -ALA is suppressed, when LA is added to the nutrient medium at low pH (pH 5), although V-uptake into the algal cells is not disturbed by LA. As demonstrated in culture media with various nitrogen sources (urea, partially hydrolized urea, ammonium salts), the development of the pH during the cultivation time is important for the presentation of the V-effect on -ALA. It is suggested that vanadium acts as a catalyst in the conversion of 4,5-dioxovaleric acid to -ALA by transamination.Abbreviations -ALA -aminolevulinic acid - LA levulinic acid - DOVA 4,5-dioxovaleric acid     

Feeding level and individual metabolic rate affect δ13C and δ15N values in carp: implications for food web studies

Stable isotope analyses are often used to calculate relative contributions of multiple food sources in an animals diet. One prerequisite for a precise calculation is the determination of the diet-tissue fractionation factor. Isotopic ratios in animals are not only affected by the composition of the diet, but also by the amount of food consumed. Previous findings regarding the latter point are controversial. As stable isotope analyses have often been used to investigate aquatic food webs, an experiment with carp (Cyprinus carpio L.) was carried out to test the influence of the feeding level and individual metabolic rate on ¹³C and ¹⁵N values of the whole body. After an initial phase, 49 carp were assigned randomly to four groups and fed the same diet at different levels for 8 weeks. For 15 fish, the energy budget was determined by indirect calorimetry. Feed and individual fish were analysed for their proximate composition, gross energy content and ¹³C and ¹⁵N values. ¹³C and ¹⁵N values differed significantly at different feeding levels. While ¹³C values of the lipids and ¹⁵N values decreased with increasing feeding rate, ¹³C values of the lipid-free matter showed a non-linear pattern. Data obtained from fish held in the respirometric system revealed a relationship between ¹³C values and the percentage retention of metabolizable energy. Our results show that reconstructing the diets of fish from the isotopic ratios when the feeding level and individual metabolic rates are unknown would introduce an error into the data used for back-calculation of up to 1 for both ¹³C and ¹⁵N values and may have substantial effects on the results of calculated diets. As other workers have pointed out, the development and application of stable isotopes to nutritional ecology studies is a field in its infancy and gives rise to erroneous, misleading results without nutritional, physiological and ecological knowledge.     

δ and κ Opiate receptors in primary astroglial cultures from rat cerebral cortex

The effects of , , and receptor-agonists on forskolin stimulated cyclic adenosine-3, 5-monophosphate (cAMP) formation were examined in astroglial enriched primary cultures from the cerebral cortex of newborn rats. Intracellular cAMP accumulation was quantified by radioimmunoassay. Morphine was used as a -receptor agonist, D-Ala-D-Leu-Enkephalin (DADLE) as a -receptor agonist and dynorphine 1–13 (Dyn) as a -receptor agonist. Basal cAMP levels were unaffected by either the opiate agonists or the antagonists used. In the presence of the cAMP stimulator forskolin, morphine had no significant effect on the cytoplasmic cAMP levels. DADLE caused a dose related inhibition of the forskolin stimulated cAMP accumulation. The effects of this receptor stimulation was blocked with the selective antagonist ICI 174.864. In the presence of Dyn, the forskolin stimulated cAMP accumulation was inhibited in a dose related manner. This receptor stimulation was blocked with the selective antagonist MR 2266. Co-administration of DADLE and Dyn resulted in a non additive inhibition of the forskolin stimulated accumulation of cAMP. These findings indicate that astroglial enriched cultures from the cerebral cortex of rats express and -receptors co-localized ont he same population of cells, and that these receptors are inhibitory coupled to adenylate cyclase.     

Stable isotope and radiocarbon compositions of methane emitted from tropical rice paddies and swamps in Southern Thailand

Stable isotopes (¹³C, D) and radiocarbon weremeasured in methane bubbles emitted from rice paddies and swamps in southernThailand. Methane emitted from the Thai rice paddies was enriched in¹³C (mean ¹³C; –51.5 ±7.1 and–56.5 ± 4.6 for mineral soil and peat soil paddies,respectively)relative to the reported mean value of methane from temperate rice paddies(– 63 ± 5). Large seasonal variation was observed in¹³C(32) in the rice paddies, whereas variationinD was much more smaller (20), indicating that variation in¹³C is due mainly to changes in methane production pathways.Values of ¹³C were lower in swamps (–66.1 ±5.1)than in rice paddies. The calculated contribution of acetate fermentation from¹³C value was greater in rice paddies (mineral soils:62–81%, peat soils: 57–73%) than in swamps (27–42%). Din methane from Thai rice paddies (–324± 7 (n=46)) isrelativelyhigher than those from 14 stations in Japanese rice paddies ranging from–362 ± 5 (Mito: n=2) to –322 ± 8(Okinawa: n=3), due tohigher D in floodwaters. ¹⁴C content in methane produced fromThai rice paddies (127±1 pMC) show higher ¹⁴Cactivity compared with previous work in paddy fields and those from Thai swamps(110±2 pMC).     

Food web structure in the high Arctic Canada Basin: evidence from δ13C and δ15N analysis

The food-web structure of the Arctic deep Canada Basin was investigated in summer 2002 using carbon and nitrogen stable isotope tracers. Overall food-web length of the range of organisms sampled occupied four trophic levels, based on 3.8 trophic level enrichment (¹⁵N range: 5.3–17.7). It was, thus, 0.5–1 trophic levels longer than food webs in both Arctic shelf and temperate deep-sea systems. The food sources, pelagic particulate organic matter (POM) (¹³C=–25.8, ¹⁵N=5.3) and ice POM (¹³C=–26.9, ¹⁵N=4.1), were not significantly different. Organisms of all habitats, ice-associated, pelagic and benthic, covered a large range of ¹⁵N values. In general, ice-associated crustaceans (¹⁵N range 4.6–12.4, mean 6.9) and pelagic species (¹⁵N range 5.9–16.5, mean 11.5) were depleted relative to benthic invertebrates (¹⁵N range 4.6–17.7, mean 13.2). The predominantly herbivorous and predatory sympagic and pelagic species constitute a shorter food chain that is based on fresh material produced in the water column. Many benthic invertebrates were deposit feeders, relying on largely refractory material. However, sufficient fresh phytodetritus appeared to arrive at the seafloor to support some benthic suspension and surface deposit feeders on a low trophic level (e.g., crinoids, cumaceans). The enriched signatures of benthic deposit feeders and predators may be a consequence of low primary production in the high Arctic and the subsequent high degree of reworking of organic material.     

Shoot δ15N correlates with genotype and salt stress in barley

Given a uniform N source, the ¹⁵N of barley shoots provided a genotypic range within treatments and a separation between control and salt-stress treatments as great as did ¹³C^*. Plant ¹⁵N has been represented in the literature as a bioassay of external source ¹⁵N and used to infer soil N sources, thus precluding consideration of the plant as a major cause in determining its own 8¹⁵N. We believe this to be the first report of plant ¹⁵N as a genetic trait. No mechanistic model is needed for use of ¹⁵N as a trait in controlled studies; however, a qualitative model is suggested for further testing.Symbol ¹⁵N (or ¹³C) the difference between: (1) the ratio of heavy to light isotopes of the element in a sample and (2) that of its reference standard     

The Genes Encoding Black Widow Spider Neurotoxins are Intronless

The overlapping fragments of the chromosomal DNA from black widow spider Latrodectus mactans carrying genes for high-molecular-mass protein neurotoxins, - and -latroinsectotoxins (-LIT and -LIT) and -latrotoxin (-LTX), were PCR-amplified and cloned. Restriction analysis of the PCR products showed that the distribution and sizes of the restriction fragments coincided with those deduced from the earlier sequencing of cDNAs of the corresponding genes. It thus followed that the -LIT and -LIT genes are intronless. Along with our data on the structure of the -latrocrustotoxin (-LCT), this implies that the lack of introns is a common feature of the black widow spider genes encoding high molecular mass neurotoxins.     

Stable oxygen isotopes in<Emphasis Type="Italic"> Porites</Emphasis> corals monitor weekly temperature variations in the northern Gulf of Aqaba,Red Sea

In order to assess the ability of Porites corals to accurately record environmental variations, high-resolution (weekly/biweekly) coral ¹⁸O records were obtained from four coral colonies from the northern Gulf of Aqaba, which grew at depths of 7, 19, 29, and 42 m along one transect. Adjacent to each colony, hourly temperatures, biweekly salinities, and monthly ¹⁸O of seawater were continuously recorded over a period of 14 months (April 1999 to June 2000). Contrary to water temperature, which shows a regular and strong seasonal variation and change with depth, seawater ¹⁸O exhibits a weak seasonality and little change with depth. Positive correlations between seawater ¹⁸O and salinity were observed. The two parameters were related to each other by the equation ¹⁸O _Seawater (, VSMOW) = 0.281 × Salinity – 9.14. The high-resolution coral ¹⁸O records from this study show a regular pattern of seasonality and are able to capture fine details of the weekly average temperature records. They resolve more than 95% of the weekly average temperature range. On the other hand, attenuation and amplification of coral seasonal amplitudes were recorded in deep, slow-growing corals, which were not related to environmental effects (temperature and/or seawater ¹⁸O) or sampling resolution. We propose that these result from a combined effect of subannual variations in extension rate and variable rates of spine thickening of skeletal structures within the tissue layer. However, no smoothing or distortion of the isotopic signals was observed due to calcification within the tissue layer in shallow-water, fast-growing corals. The calculations from coral ¹⁸O calibrations against the in situ measurements show that temperature (T) is related to coral ¹⁸O ( _c ) and seawater ¹⁸O ( _w ) by the equation T (°C) = –5.38 ( _c – _w ) –1.08. Our results demonstrate that coral ¹⁸O from the northern Gulf of Aqaba is a reliable recorder of temperature variations, and that there is a minor contribution of seawater ¹⁸O to this proxy, which could be ignored.     

Variations in the natural abundance of 15N in ryegrass/white clover shoot material as influenced by cattle grazing

Biological N2 fixation in clover is an important source of N in low external-N input farming systems. Using the natural 15N-abundance method, variations in N2 fixation were investigated in grazed and mowed plots of a ryegrass/white clover field. Ryegrass 15N varied considerably, from 0.2 to 5.6 under mowed conditions and from –3.3 to 11.6 under grazed conditions. Variations in 15N white clover were lower than in ryegrass, especially in the mowed plots (SE = 0.05, n = 20). The variations in the percentage of nitrogen derived from the atmosphere (%Ndfa) in white clover were highest in the grazed plots where it ranged from 12 to 96% (mean = 64%) compared with the mowed plots where it ranged from 64 to 92% (mean = 79%). Thus, the N2 fixation per unit white clover DM in the grazed ley was lower and more variable than under mowing conditions.Urine from dairy cows equivalent to 0, 200, 400 and 800 kg N ha-1 was applied to a ryegrass/white clover plot 6, 4 or 2 weeks before harvest. Without urine application 15N of ryegrass was positive. By increasing urine application (15N = –1) two weeks before sampling, the 15N of ryegrass decreased strongly to about –7 (P < 0.001). However, this effect was only observed when urine was applied two weeks before sampling. When applying 800 kg N four and six weeks before sampling, 15N in ryegrass was not significantly different from the treatment without urine application. White clover 15N was unaffected by whatever changes occurred in 15N of the plant-available soil N pool (reflected in 15N of ryegrass). This indicates that within the time span of this experiment, N2 fixation per unit DM was not affected by urine. Therefore, newly deposited urine may not be the main contributing factor to the variation in %Ndfa found in the grazed fields. This experiment suggested that the natural abundance method can be applied for estimating %Ndfa without disturbance in natural animal-grazed systems.     

Using stable isotopes to investigate migratory connectivity of the globally threatened aquatic warbler Acrocephalus paludicola

Understanding the links between breeding and wintering areas of migratory species has important ecological and conservation implications. Recently, stable isotope technology has been used to further our understanding. Stable isotope ratios vary geographically with a range of biogeochemical factors and isotope profiles in organisms reflect those in their food and environment. For inert tissues like feathers, isotope profiles reflect the environment in which they were formed. Following large-scale habitat destruction, the globally threatened aquatic warbler Acrocephalus paludicola has a fragmented breeding population across central Europe, largely in Belarus, Poland and Ukraine. The species sub-Saharan African wintering grounds have not yet been discovered, and this significantly hampers conservation efforts. Aquatic warblers grow their flight feathers on their wintering grounds, and we analysed stable isotope ratios (¹⁵N, ¹³C, D) in rectrices of adults from six main breeding sites (subpopulations) across Europe to determine whether different breeding subpopulations formed a single mixed population on the wintering grounds. ¹⁵N varies considerably with dietary trophic level and environmental factors, and D with the D in rainfall; neither varied between aquatic warbler subpopulations. Uniform feather ¹⁵N signatures suggest no major variation in dietary trophic level during feather formation. High variance and inter-annual differences in mean D values hinder interpretation of these data. Significant differences in mean ¹³C ratios existed between subpopulations. We discuss possible interpretations of this result, and consider differences in moulting latitude of different subpopulations to be the most parsimonious. ¹³C in plants and animals decreases with latitude, along a steep gradient in sub-Saharan Africa. Birds from the most north-westerly breeding subpopulation (Karsibor, Poland) had significantly lower variance in ¹³C and ¹⁵N than birds from all other sites, suggesting either that birds from Karsibor are less geographically dispersed during moult, or moult in an area with less isotopic heterogeneity. Mean ¹³C signatures from winter-grown feathers of different subpopulations were positively correlated with the latitude and longitude of breeding sites, suggesting a strong relationship between European breeding and African winter moulting latitudes. The use of stable isotopes provides novel insights into migratory connectivity and migration patterns in this little-known threatened species.