Radical changes in the Wadden Sea fauna and flora over the last 2,000 years

Humans have interacted with the Wadden Sea since its origin 7,500 years ago. However, exploitation, habitat alteration and pollution have strongly increased since the Middle Ages, affecting abundance and distribution of many marine mammals, birds, fish, invertebrates and plants. Large whales and some large birds disappeared more than 500 years ago. Most small whales, seals, birds, large fish and oysters were severely reduced by the late 19th and early 20th centuries, leading to the collapse of several traditional fisheries. In the 20th century, conservation efforts have enabled some breeding birds and seals to recover. But other species declined further due to continuing exploitation, habitat destruction, pollution and eutrophication. Moreover, complex three-dimensional habitats such as oyster banks, Sabellaria reefs and subtidal eelgrass beds have been lost completely. In contrast, several opportunistic species such as gulls, polychaetes, green algae and exotic invaders increased during the 20th century. Taken together, multiple human impacts have caused dramatic losses of large predators and habitat-building species in the Wadden Sea over the last 500 years. Although still of high natural value and global importance, the Wadden Sea is a fundamentally changed ecosystem. On the other hand, reduced hunting pressure, increased habitat protection and reduced river pollution have enabled the recent recovery of several species and an increase in environmental quality. These successes, together with a historical vision of what was once possible, should guide current and future conservation, restoration and management efforts towards a more sustainable interaction between man and the sea.     

Insulin induces changes in the subcellular distribution of actin and 5′-nucleotidase

The present study utilized a cultured adult myocardial cell model to examine the arachidonic acid metabolism under different cell-damaging and normoxic conditions. Cell injury was caused by short-time hypoxia, calcium ionophore A 23187-triggered cell-damage under hypoxia and cell disruption by freezing and thawing. The current study demonstrates that under the cell-damaging conditions cultured adult heart myocytes resemble myocardial cells under normoxic conditions in metabolizing arachidonic acid into triacylglycerols and phospholipids as the major route (a), in formation of ETYA-inhibitable indomethacin-resistant lipid metabolites in minor amounts (b) and in being independent of calcium overload in the metabolic pathways of arachidonic acid metabolism (c). The ETYA-inhibitable components were resolved by HPLC. There was no evidence in formation of lipoxygenase products. The results were supported by negative hybridisation experiments of the total mRNA isolated from adult myocardial cells with a cDNA probe of a red-cell-specific lipoxygenase mRNA. We conclude from these observations that cell injury does not result in expression of lipoxygenase activities in heart myocytes.Abbreviations HETE Hydroxyeicosatetraenoic acid - DiHETE Dihydroxyeicosatetraenoic acid - ETYA 5.8.11.14-Eicosatetraynoic acid - TLC Thin-Layer Chromatography - NP-HPLC Normal Phase-High Performance Liquid Chromatography - RBC Red Blood Cell - LOX Lipoxygenase     

Complexity and developmental changes in the expression pattern of claudins at the blood–CSF barrier

The choroid plexus epithelium controls the movement of solutes between the blood and the cerebrospinal fluid. It has been considered as a functionally more immature interface during brain development than in adult. The anatomical basis of this barrier is the interepithelial choroidal junction whose tightness has been attributed to the presence of claudins. We used quantitative real-time polymerase chain reaction, Western blot and immunohistochemistry to identify different claudins in the choroid plexuses of developing and adult rats. Claudin-1, -2, and -3 were highly and selectively expressed in the choroid plexus as compared to brain or parenchyma microvessels and were localized at epithelial junctions. Claudin-6, -9, -19, and -22 also displayed a previously undescribed choroidal selectivity, while claudin-4, -5, and -16 were enriched in the cerebral microvessels. The choroidal pattern of tight junction protein expression in prenatal brains was already complex and included occludin and zonula occludens proteins. It differed from the adult pattern in that the pore-forming claudin-2, claudin-9, and claudin-22 increased during development, while claudin-3 and claudin-6 decreased. Claudin-2 and claudin-11 presented a mirror image of abundance between lateral ventricle and fourth ventricle choroid plexuses. Imunohistochemical analysis of human fetal and postnatal brains for claudin-1, -2, and -3 demonstrated their early presence and localization at the apico-lateral border of the choroid plexus epithelial cells. Overall, choroidal epithelial tight junctions are already complex in developing brain. The observed differences in claudin expression between developing and adult choroid plexuses may indicate developmental differences in selective blood–cerebrospinal fluid transport functions.     

Surface potential changes in lipid monolayers and the ‘cut-off’ in anaesthetic effects of N-alkanols

The effects of 0.09 saturated solutions of the n-alkanols n-hexanol to n-tridecanol on the surface (compensation) potential of lipid monolayers have been examined. Actions on monolayers spread from pure egg phosphatidylcholine have been compared with effects on a system containing a 2:1 mole ratio of egg phosphatidylcholine and cholesterol. The mean compensation potential for the pure phospholipid system was 475 ± 9 mV; addition of cholesterol increased the potential to 503 ± 10 mV. All n-alkanols tested reduced the surface potential in both systems. The reduction was larger in the pure phospholipid system but the difference in effect between lipid systems declined as the n-alkanol chainlength increased, becoming negligible by n-tridecanol. These results are considered in relation to the ‘cut-off’ in biological activity of n-alkanols around n-tridecanol.     

Are the short-photoperiod-induced decreases in serum prolactin responsible for the seasonal changes in energy balance in Syrian and Siberian hamsters?

Serum prolactin (PRL) decreases in Syrian (Mesocricetus auratus) and Siberian (Phodopus sungorus sungorus) hamsters following short-photoperiod exposure. Both species also exhibit short-photoperiod-induced changes in body and lipid mass, but in opposite directions; Syrian hamsters increase and Siberian hamsters decrease their body weight, changes reflected nearly exclusively in their carcass lipid content. The purpose of these experiments was to determine whether the photoperiod-induced changes in PRL were responsible for the seasonal changes in energy balance in Syrian and Siberian hamsters by using the strategy of experimentally producing serum PRL levels opposite to those normally associated with the photoperiod in which the animals were housed. In long photoperiods serum PRL was reduced to short-day levels by subcutaneous (s.c.) CB-154 (bromoergocryptine, a dopamine agonist) injections. In short photoperiods, serum PRL was elevated to long-day levels in Syrian hamsters by ectopic pituitary explants, and in Siberian hamsters, serum PRL was elevated by chronic s.c. infusions of ovine PRL (oPRL). In both species, manipulations of serum PRL did not affect food intake, carcass composition, or the wet weight of various white and brown adipose tissue pads (WAT and BAT, respectively). Body weight increased in CB-154-treated Syrian hamsters and decreased in Siberian hamsters, an effect partially reversed by coadministration of oPRL in Syrian, but not Siberian, hamsters. Thus, lowering serum PRL to short-day levels in long-day-housed hamsters of both species mimicked the directional change in body weight appropriate for each species when they are exposed to short days. This effect of CB-154 on body weight may be a result of some as yet unidentified effect of dopaminergic stimulation on overall growth since 1) these changes in body weight were not reflected as changes in lipid mass, as occurs naturally following short-day exposure for each species, and 2) neither species exhibited a reciprocal change in body weight when serum PRL was experimentally elevated in short days. Alternatively, it may be that once the energetic responses to short-day exposure have been fully expressed, the ability of PRL to stimulate the target sites of action for PRL for these responses may be decreased. BAT protein content, cytochrome oxidase activity (measures of metabolic growth of this tissue), and retroperitoneal total and specific lipoprotein lipase (LPL) activities were increased by CB-154 treatment in Syrian hamsters.(ABSTRACT TRUNCATED AT 400 WORDS)     

Do changes in gene expression contribute to sexual isolation and reinforcement in the house mouse?

Expression divergence, rather than sequence divergence, has been shown to be important in speciation, particularly in the early stages of divergence of traits involved in reproductive isolation. In the two European subspecies of house mice, Mus musculus musculus and Mus musculus domesticus, earlier studies have demonstrated olfactory‐based assortative mate preference in populations close to their hybrid zone. It has been suggested that this behaviour evolved following the recent secondary contact between the two taxa (~3,000 years ago) in response to selection against hybridization. To test for a role of changes in gene expression in the observed behavioural shift, we conducted a RNA sequencing experiment on mouse vomeronasal organs. Key candidate genes for pheromone‐based subspecies recognition, the vomeronasal receptors, are expressed in these organs. Overall patterns of gene expression varied significantly between samples from the two subspecies, with a large number of differentially expressed genes between the two taxa. In contrast, only ~200 genes were found repeatedly differentially expressed between populations within M. m. musculus that did or did not display assortative mate preferences (close to or more distant from the hybrid zone, respectively), with an overrepresentation of genes belonging to vomeronasal receptor family 2. These receptors are known to play a key role in recognition of chemical cues that handle information about genetic identity. Interestingly, four of five of these differentially expressed receptors belong to the same phylogenetic cluster, suggesting specialization of a group of closely related receptors in the recognition of odorant signals that may allow subspecies recognition and assortative mating.     

Volume changes and electrostriction in the primary photoreactions of various photosynthetic systems: estimation of dielectric coefficient in bacterial reaction centers and of the observed volume changes with the Drude–Nernst equation

Photoacoustics (PA) allows the determination of enthalpy and volume changes of photoreactions in photosynthetic reaction centers on the 0.1–10 μs time scale. These include the bacterial centers from Rb. sphaeroides, PS I and PS II centers from Synechocystis and in whole cells. In vitro and in vivo PA data on PS I and PS II revealed that both the volume change (–26 A³) and reaction enthalpy (–0.4 eV) in PS I are the same as those in the bacterial centers. However the volume change in PS II is small and the enthalpy far larger, –1 eV. Assigning the volume changes to electrostriction allows a coherent explanation of these observations. One can explain the large volume decrease in the bacterial centers with an effective dielectric coefficient of ∼4. This is a unique approach to this parameter so important in estimation of protein energetics. The value of the volume contraction for PS I can only be explained if the acceptor is the super- cluster (Fe₄S₄)(Cys₄) with charge change from –1 to –2. The small volume change in PS II is explained by sub-μs electron transfer from Y_Z anion to P₆₈₀ cation, in which charge is only moved from the Y_Z anion to the Q_A with no charge separation or with rapid proton transfer from oxidized Y_Z to a polar region and thus very little change in electrostriction. At more acid pH equally rapid proton transfer from a neighboring histidine to a polar region may be caused by the electric field of the P₆₈₀ cation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Polyphenol gene expression and changes in anthocyanins and polyphenols in the skin of ‘Braeburn’ apples after the autumn application of prohexadione-calcium

Prohexadione-calcium (Pro-Ca) transient inhibits 2-Oxoglutarate-dependent dioxygenases and causes significant changes in the flavonoid spectrum of apple. In the present study the influence of two autumn preharvest applications of Pro-Ca on the polyphenol metabolism in apple peel during the advanced maturation was investigated. Pro-Ca was sprayed in two doses, approximately five and 3 weeks before the technological maturity. Changes in the concentrations of hydroxycinnamic acids, dihydrochalcones, flavonols, flavanols and anthocyanins as well as their related gene expression and enzyme activities in the apple peel were monitored six times during the advanced maturation until the technological maturity of the fruits. To evaluate its influence on red coloration differences in the chromatic values a*, h° and L* between the treated and untreated apples were monitored. The parameters showed a temporary effect of Pro-Ca on the intensity of red coloration, which was not detected anymore at the technological maturity of apples. The application of Pro-Ca decreased the flavanone 3-hydroxylase activity and slightly inhibited activities of all the enzymes analyzed. Concomitantly, the concentrations of anthocyanins in the peel of the treated apples decreased, whereas the concentrations of hydroxycinnamic acids, dihydrochalcones and flavan 3-ols increased. Flavonol concentrations, however, remained unchanged. The expression of ANS, ANR, FGT and MYB10 was downregulated after the Pro-Ca treatment. The results indicate that the autumn application of Pro-Ca modulates the biosynthetic pathway resulting in distinct changes in the flavonoid composition in the apple peel of ‘Braeburn’ apples. However, the changes are temporary and are generally suspended during apple storage.     

Ontogenetic changes in the predator–prey interactions between threadsail filefish and moon jellyfish

The global climate change may lead to more extreme climate events such as severe flooding creating excessive pulse-loading of nutrients, including nitrogen (N), to freshwaters. We conducted a 3-month mesocosm study to investigate the responses of phytoplankton, zooplankton and Vallisneria spinulosa to different N loading patterns using weekly and monthly additions of in total 14 g N m^?2 month^?1 during the first 2 months. The monthly additions led to higher phytoplankton chlorophyll a and total phytoplankton biomass than at ambient conditions as well as lower leaf biomass and a smaller ramet number of V. spinulosa. Moreover, the biomass of cyanobacteria was higher during summer (August) in the monthly treatments than those with weekly or no additions. However, the biomass of plankton and macrophytes did not differ among the N treatments at the end of the experiment, 1 month after the termination of N addition. We conclude that by stimulating the growth of phytoplankton (cyanobacteria) and reducing the growth of submerged macrophytes, short-term extreme N loading may have significant effects on shallow nutrient-rich lakes and that the lakes may show fast recovery if they are not close to the threshold of a regime shift from a clear to a turbid state.     

Proteome changes in the myocardium of experimental chronic diabetes and hypertension: role of PPARα in the associated hypertrophy

Diabetes with or without the presence of hypertension damages the heart. However, there is currently a lack of information about these associated pathologies and the alteration of linked proteins. For these reasons, we were interested in the potential synergistic interaction of diabetes and hypertension in the heart, focusing on the proteome characterization of the pathological phenotypes and the associated hypertrophic response. We treated normotensive and spontaneously hypertensive (SHR) rats with either streptozotocin or vehicle. After 22 weeks, type-I diabetic (DM1), SHR, SHR/DM1 and control left-ventricles were studied using proteomic approaches. Proteomics revealed that long-term DM1, SHR and SHR/DM1 rats exhibited 24, 53 and 53 altered proteins in the myocardia, respectively. DM1 myocardium showed over-expression of apoptotic and cytoskeleton proteins, and down-regulation of anti-apoptotic and mitochondrial metabolic enzymes. In both SHR and SHR/DM1 these changes were exacerbated and free fatty-acid (FFA) ß-oxidation enzymes were additionally decreased. Furthermore, SHR/DM1 hearts exhibited a misbalance of specific pro-hypertrophic, anti-apoptotic and mitochondrial ATP-carrier factors, which could cause additional damage. Differential proteins were validated and then clustered into different biological pathways using bioinformatics. These studies suggested the implication of FFA-nuclear receptors and hypertrophic factors in these pathologies. Although key ß-oxidation enzymes were not stimulated in DM1 and hypertensive hearts, peroxisome proliferator-activated receptors-α (PPARα) were potentially activated for other responses. In this regard, PPARα stimulation reduced hypertrophy and pro-hypertrophic factors such as annexin-V in high-glucose and angiotensin-II induced cardiomyocytes. Thus, activation of PPARα could reflect a compensatory response to the metabolic-shifted, apoptotic and hypertrophic status of the hypertensive-diabetic cardiomyopathy.     

Site of action for β-endorphin-induced changes in plasma luteinizing hormone and prolactin in the ovariectomized rat

A number of sites have been hypothesized as loci at which opioid substances act to alter the secretion of luteinizing hormone (LH) and prolactin (PRL) (1–8). The aim of the present study was to determine the site(s) at which the opioid peptide β-endorphin (β-END) acts to influence plasma LH and PRL levels in the ovariectomized (OVX) rat. β-END, administered into the third ventricle of conscious OVX rats fitted with jugular catheters, significantly decreased plasma LH in doses ? 50 ng and increased PRL levels at all doses administered (10, 50, 100 and 250 ng) in a dose dependent fashion. To identify possible central nervous system sites of action, 250 ng β-END was unilaterally infused into various brain sites. Plasma LH was significantly decreased and plasma PRL significantly increased by infusions into the ventromedial hypothalamic area, the anterior hypothalamic area, and the preoptic-septal area. There was no significant effect of β-END infusions into the lateral hypothalamic area, amygdala, midbrain central gray, or caudate nucleus. When hemipituitaries of OVX rats were incubated $\text{in}$$\text{vitro}$ with β-END (10^?7M to 10^?5M), there was no suppression of basal or LHRH-induced LH release, nor was there any alteration of basal PRL release. It is concluded that β-END acts at a medial hypothalamic and/or preoptic-septal site and not the pituitary, to alter secretion of LH and PRL.     

High temperature — induced changes in germination,seedling vigour and the metabolic activities in rice seeds

The effects of temperature treatments on rice seeds in the early imbibitional phase were studied with respect to changes in germination percentage, seedling growth in terms of root and shoot lengths, water uptake, respiration and hydrolytic enzymes,viz. α-amylase, adenosine triphosphatase (ATPase) and phytase. As compared with the control, the treatment for 30 min at 50 °C caused a variable degree of increase in all these characters and a positive correlation existed between seedling vigour and hydrolase activities. The treatment for 30 min at 60 °C, on the other hand, elicited a retarding influence on these characters. As regards the enzyme activities, the damaging effect of 60 °C could, however, be visualized only after 72 h of germination which was preceded by an enhancement during the early hours.     

Developmental changes in functional expression and β-adrenergic regulation of If in the heart of mouse embryo

The hyperpolarization-activated current (If) plays an important role in determining the spontaneousrate of cardiac pacemaker cells. The automatic rhythmicity also exists in working cells of embryonic heart,therefore we studied developmental changes in functional expression and β-adrenergic regulation of If inembryonic mouse heart. The expression of If is high in early developmental stage (EDS) (10.5 d after coitus)ventricular myocytes, low in intermediate developmental stage (IDS) (13.5 d) atrial or ventricular myocytesand even lower in late developmental stage (LDS) (16.5 d) atrial or ventricular myocytes, indicating thatthese cells of the EDS embryonic heart have some properties of pacemaker cells. β-adrenergic agonistisoproterenol (ISO) stimulates If in LDS but not in EDS cardiomyocytes, indicating that theβ-adrenergicregulation of If is not mature in EDS embryonic heart. But forskolin (a direct activator of adenylate cyclase)and 8-Br-cAMP (a membrane-permeable analogue of cAMP) increase the amplitude of If in EDS cells,indicating that adenylate cyclase and cAMP function fairly well at early stage of development. Furthermore,the results demonstrate that If is modulated by phosphorylation via cAMP dependent PKA both in EDSand LDS cells.     

Developmental changes in functional expression and β-adrenergic regula-tion of If in the heart of mouse embryo

The hyperpolarization-activated current (If) plays an important role in determining the spontaneous rate of cardiac pacemaker cells. The automatic rhythmicity also exists in working cells of embryonic heart,therefore we studied developmental changes in functional expression and β-adrenergic regulation of If in embryonic mouse heart. The expression of If is high in early developmental stage (EDS) (10.5 d after coitus) ventricular myocytes, low in intermediate developmental stage (IDS) (13.5 d) atrial or ventricular myocytes and even lower in late developmental stage (LDS) (16.5 d) atrial or ventricular myocytes, indicating that these cells of the EDS embryonic heart have some properties of pacemaker cells, β-adrenergic agonist isoproterenol (ISO) stimulates If in LDS but not in EDS cardiomyocytes, indicating that the β-adrenergic regulation of If is not mature in EDS embryonic heart. But forskolin (a direct activator of adenylate cyclase) and 8-Br-cAMP (a membrane-permeable analogue of cAMP) increase the amplitude of If in EDS cells,indicating that adenylate cyclase and cAMP function fairly well at early stage of development. Furthermore,the results demonstrate that If is modulated by phosphorylation via cAMP dependent PKA both in EDS and LDS cells.     

Developmental changes in functional expression andβ-adrenergic regulation of I_f in the heart of mouse embryo

The hyperpolarization-activated current (If) plays an important role in determining the spontaneous rate of cardiac pacemaker cells. The automatic rhythmicity also exists in working cells of embryonic heart, therefore we studied developmental changes in functional expression and β-adrenergic regulation of Iy in embryonic mouse heart. The expression of If is high in early developmental stage (EDS) (10.5 d after coitus) ventricular myocytes, low in intermediate developmental stage (IDS) (13.5 d) atrial or ventricular myocytes and even lower in late developmental stage (LDS) (16.5 d) atrial or ventricular myocytes, indicating that these cells of the EDS embryonic heart have some properties of pacemaker cells.β-adrenergic agonist isoproterenol (ISO) stimulates If in LDS but not in EDS cardiomyocytes, indicating that the β-adrenergic regulation of If is not mature in EDS embryonic heart. But forskolin (a direct activator of adenylate cyclase) and 8-Br-cAMP (a membrane-permeable analogue of cAMP) increase t     

Hormonal changes in spontaneous and aglépristone-induced parturition in dogs

To increase our understanding of the endocrine changes associated with parturition in dogs, plasma concentrations of progesterone (P4), 15-ketodihydroprostaglandin F(2alpha) (PGFM), estradiol-17-beta (E2beta), cortisol, ACTH, prolactin (PRL), LH, and FSH were measured in six spontaneously whelping bitches and in six bitches in which parturition was induced with the progesterone-receptor blocker aglépristone on day 58 of pregnancy. Expulsion of pups in the induced group took place in the presence of P4 concentrations that were still elevated. PGFM concentrations increased before parturition in both groups, but levels were lower in the induced bitches. PGFM levels reached a maximum in both groups during parturition and quickly decreased in the spontaneously whelping group after parturition, but remained elevated in the induced group. In both groups, cortisol concentrations reached similar maximum levels during the last 30 h before the onset of expulsion. During the 3 days postpartum, cortisol concentrations were higher in the induced group. The highly variable ACTH concentrations did not differ significantly throughout the study within or between groups. In both groups, E2beta concentrations decreased and PRL concentrations increased between the late gestational period and the 30-h period before parturition. Concentrations of both LH (spontaneously whelping group) and FSH (both groups) decreased between late gestation and the postpartum period. The results of this study illustrate the hormonal changes around parturition in the bitch, and reveal that aglépristone-induced parturition is associated with still incomplete luteolysis, an altered PGFM profile, and elevated postpartum cortisol concentrations as compared with spontaneously whelping dogs.