Step-wise change of Asian interior climate preceding the Eocene–Oligocene Transition (EOT)

Understanding the global climate change from greenhouse to icehouse conditions at the Eocene–Oligocene Transition (EOT) 34 million years ago requires climatic records from oceanic as well as continental realms of the key Late Eocene “doubthouse” period preceding this switch. Here, we report integrated stratigraphic results from well-dated Late Eocene continental mudflat to saline lake paleoenvironments of the Xining Basin (northeastern Tibetan Plateau, western China) recording regional and global change. Cyclostratigraphic analysis strongly suggests continuous dominance of the 41-kyr obliquity cycle in the whole late Eocene interval down to the base of polarity chron C18n.2n at 39 Ma with additional input of the ~ 100-kyr eccentricity cycle up to the base of chron C13r at ~ 34.7 Ma. This might imply that high-latitude climates dominated the area long before the EOT, probably related to incipient ice-volume fluctuations. Furthermore, our results reveal two paleoenvironmental deterioration steps preceding the Eocene–Oligocene Transition. The first step occurs in the top of chron C17n.1n at ~ 36.6 Ma. This age closely corresponds to (1) the high-altitude pollen appearance in chron C16.2r at ~ 36.4 Ma in the same section, (2) the recently dated final retreat of the Tarim Sea in western China, and (3) a shift from precession to obliquity dominance in the Atlantic Ocean. This near co-occurrence suggests global change at this time. We hypothesize this change is related to an increase in incipient ice sheet volume leading to passing threshold conditions for the high-altitude pollen appearance and Tarim Sea retreat, finally leading to decreased moisture availability in the Xining Basin. At the second step, in the base of chron C13r at ~ 34.7 Ma, a substantial increase in clastic sedimentation rates is observed. This might relate to increased climate variability preceding the greenhouse to icehouse transition at the EOT that prevented landscapes to attain equilibrium configurations.     

Late Miocene paleoenvironmental changes in North Africa and the Mediterranean recorded by geochemical proxies (Monte Gibliscemi section,Sicily)

The astronomically tuned marls of the Monte Gibliscemi section, Sicily, constitute an archive to trace the late Miocene palaeoenvironmental conditions (~ 9.7–7.0 Ma) in North Africa. Here we have utilised carbonate content and Al-normalised geochemical proxies to trace changes in terrigenous source area and bottom-water ventilation. The terrigenous input into the section is dominated by North African river systems draining into the Eastern Mediterranean. The proxy parameters indicate that the palaeoenvironmental conditions in North Africa were humid from 9.5 Ma onward with high fluvial input to the Mediterranean. Increases in the Si/Al and Mg/Al ratios occurred from 8.4 to 8.2 Ma and from 8.05 to 7.75 Ma, with maximum values similar to those of Messinian diatomites. These peaks indicate conditions of enhanced biosiliceous productivity and the presence of authigenic clay formation. Sluggish water circulation in the Mediterranean during those times is inferred from the Mn/Al and V/Al behaviour. Late Miocene changes of the Betic (southern Spain) and Rifian (Morocco) Mediterranean–Atlantic gateways are interpreted as the driving force for the changes in water circulation and the transgression associated with the opening of the Rifian corridor is interpreted to occur at 7.8 Ma.     

Xenon reduces activation of transient receptor potential vanilloid type 1 (TRPV1) in rat dorsal root ganglion cells and in human TRPV1-expressing HEK293 cells

AimsXenon provides effective analgesia in several pain states at sub-anaesthetic doses. Our aim was to examine whether xenon may mediate its analgesic effect, in part, through reducing the activity of transient receptor potential vanilloid type 1 (TRPV1), a receptor known to be involved in certain inflammatory pain conditions.Main methodsWe studied the effect of xenon on capsaicin-evoked cobalt uptake in rat cultured primary sensory neurons and in human TRPV1 (hTRPV1)-expressing human embryonic kidney 293 (HEK293) cells. We also examined xenon's effect on the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) in the rat spinal dorsal horn evoked by hind-paw injection of capsaicin.Key findingsXenon (75%) reduced the number of primary sensory neurons responding to the TRPV1 agonist, capsaicin (100 nM–1 μM) by ~ 25% to ~ 50%. Xenon reduced the number of heterologously-expressed hTRPV1 activated by 300 nM capsaicin by ~ 50%. Xenon (80%) reduced by ~ 40% the number of phosporylated ERK1/2-expressing neurons in rat spinal dorsal horn resulting from hind-paw capsaicin injection.SignificanceXenon substantially reduces the activity of TRPV1 in response to noxious stimulation by the specific TRPV1 agonist, capsaicin, suggesting a possible role for xenon as an adjunct analgesic where hTRPV1 is an active contributor to the excitation of primary afferents which initiates the pain sensation.     

Palynofacies analysis of the late Holocene San Nicolás terrace of the Cauca paleolake and paleohydrology of northern South America

The detailed palynofacies analysis of the late Holocene laminated succession of the San Nicolás terrace in the middle Cauca Valley, northern Colombia, is interpreted as the product of sedimentation in a lacustrine to fluvial dominated setting. Radiocarbon analyses reveal that the succession was deposited between ~ 3500 and a few hundreds of years with sedimentary rates in excess of ~ 600 cm/ka. The milimetric laminae and the high frequency fluctuations in the content of the palynodebris of terrestrial origin, i.e., the high altered ligno-cellulosic debris/amorphous organic matter ratio, together with diatoms typical of fluvial conditions, such as Aulacoseira granulata do suggest an intermittent and continuous hydrological and biological connectivity with the Cauca River, akin to a varzea lacustrine environment. Drier to wetter conditions appear to occur, moving upward in the succession. This might reflect regional precipitation conditions all along the Cauca Valley and the southern migration of the intertropical convergence zone.     

Dynamic assessment of the value of vegetation carbon fixation and oxygen release services in Qinghai Lake basin

Studies on ecosystem service function have an important significance for analyzing and understanding global warming. With the introduction of geographic information system (GIS) and remote sensing (RS) technologies for the evaluation of ecosystem service function, the scope for analysis has been widening. Increasing number of researchers use these technologies to quantify the value of ecosystem service functions and reveal their spatial-temporal variability. By using the data for the interpretation of five RS images and net primary productivity (NPP) in Qinghai Lake basin, we assessed the value of vegetation carbon fixation and oxygen release services and revealed their dynamic variation in this basin. The result suggested that the average values of vegetation carbon fixation and oxygen release services in Qinghai Lake basin between 1987 and 2010 were spatially distributed in a ring shape around the Qinghai Lake and decreased from southeastern to the north and northwestern regions; the northwestern areas had the lowest value. The vegetation carbon fixation value between 1987 and 2010 was on an average 28.87 × 10⁸ yuan/a in Qinghai Lake basin, whereas the oxygen release value was 64.41 × 10⁸ yuan/a. Alpine meadow ecosystem showed the highest value of vegetation carbon fixation and oxygen release services function in Qinghai Lake basin, with average values of 18.28 × 10⁸ yuan/a and 40.79 × 10⁸ yuan/a, respectively, followed by those of temperate steppe and sparse vegetation. The vegetation carbon fixation and oxygen release values in Qinghai Lake basin gradually increased from 1987 to 2010, with the maximum value in 2010. By the end of 2010, the values increased by 7.19 × 10⁸ yuan and 16.04 × 10⁸ yuan, respectively. The values slightly decreased in barren land, lakeside marsh, river valley swamp, and sandy areas, but increased to different degrees in other ecosystems. Among them, the largest increase was noted in alpine meadow (4.38 × 10⁸ yuan and 9.78 × 10⁸ yuan, respectively), followed by those in temperate steppe with increased values of 1.12 × 10⁸ yuan and 2.49 × 10⁸ yuan, respectively.     

Periosteal microcirculatory action of chronic estrogen supplementation in osteoporotic rats challenged with tourniquet ischemia

AimsTransient ischemia of osteoporotic bones during elective orthopedic surgery or fracture repair carries risks for serious complications, and estrogen loss or replacement has a potential to influence ischemia–reperfusion-induced inflammatory activation. To clarify this, we investigated the periosteal inflammatory changes in a clinically relevant time frame in ovariectomized rats, an experimental model of postmenopausal bone loss. Furthermore, the effects of chronic estrogen supplementation on the postischemic local and systemic inflammatory reactions were assessed.Main methodsBilateral ovariectomy or sham operation was performed in 3-month-old female Sprague–Dawley rats. Five months later, estrogen replacement therapy with 17β-estradiol (20 μg^? 1 kg^? 1 day^? 1) or vehicle treatment was initiated. The microcirculatory inflammatory consequences of 60-min total hindlimb ischemia followed by 180-min reperfusion were examined 11 months after ovariectomy and were compared with those in 3-month-old animals.Key findingsThe osteoporosis that developed 5 months after ovariectomy was significantly ameliorated by estrogen replacement therapy. Both in ovariectomized and in non-ovariectomized animals, ischemia–reperfusion elevated the neutrophil adherence ~ 3-fold in the postcapillary venules of the periosteum (intravital microscopy), with an ~ 50–60% increase in intravascular neutrophil activation (CD11b; FACS analysis), an enhanced TNF-α release (ELISA) and periosteal expression of ICAM-1 (the endothelial ligand of CD11b; immunohistochemistry). Exogenous 17β-estradiol considerably reduced TNF-α release and the number of neutrophil–endothelial interactions in the periosteum, without affecting the CD11b and ICAM-1 expression changes.SignificanceOsteoporosis itself does not increase the magnitude of the limb ischemia–reperfusion-associated periosteal inflammatory reaction. Chronic estrogen supplementation, however, reverses osteoporosis and significantly ameliorates the microcirculatory consequences of transient ischemia.     

Geopalaeontological setting,chronology and palaeoenvironmental evolution of the Baccinello-Cinigiano Basin continental successions (Late Miocene,Italy)

The Latest Miocene succession of the Baccinello-Cinigiano Basin in southern Tuscany (Italy) recorded a faunal turnover documenting the extinction of an older, insular, endemic faunal complex characterised by the extinct ape Oreopithecus bambolii and the setting of a new, continental, European faunal complex including the colobine monkey Mesopithecus. A similar turnover pattern (Late Miocene ape/Latest Miocene Cercopithecidae) is generally observed in Late Miocene continental successions of Eurasia, from Spain to central Europe, Southwest Europe, the near East, and Southwest Asia. Abundant literature reports that the Late Miocene Eurasian hominoid primate distribution closely tracks the climatic/environmental changes occurring during the 12–9 Ma interval, until their extinction in western Europe. In the primate record, the dispersion of Cercopithecidae and the contraction of hominids is interpreted as an event depicting a pattern of “continentalisation” in the Old World. The sedimentary succession of the Baccinello-Cinigiano basin, one of the longest continuous vertebrate-bearing continental successions in the Neogene Italian record, contributes to the debate on this hypothesis. This paper provides an overview of the main characteristics of the sedimentary succession, the chronological constraints (biochronology, radiometric datings, magnetostratigraphy), and the palaeoenvironmental evolution as derived from palaeobiological approaches and from the study of stable carbon and oxygen isotope contents along the entire sedimentary succession. The 2 myr geological history of the Baccinello Cinigiano Basin, which documents the evolutionary history of Oreopithecus and associated faunas, does not have a direct relation with the event of the Messinian Salinity Crisis. The evolutionary history of Baccinello-Cinigiano Basin and its palaeontological record have been mainly driven by the regional tectonism and palaeogeographic changes that affected the northern Tyrrhenian regions in Late Miocene (Latest Tortonian–Messinian) times.     

Integrated stratigraphy of the Early Miocene lacustrine deposits of Pag Island (SW Croatia): Palaeovegetation and environmental changes in the Dinaride Lake System

An integrated stratigraphic study of a Neogene lacustrine succession on the Pag Island (Croatia), combining quantitative pollen analysis, magnetostratigraphy, cyclostratigraphy, biostratigraphy and gamma-ray measurements, provides new insights into orbitally controlled variations in palaeo-vegetation and depositional patterns in the Dinaride Lake System. The quantitative palynological record shows a cyclical pattern of vegetation changes that closely corresponds to sedimentological patterns. The intervals with a high abundance of thermophilous and xeric indicators, suggesting a warm and dry climate, generally coincide with intervals of frequent lignite deposition and shallow lake facies. This suggests that both records are dominantly controlled by variations in past climatic conditions and lake level. Our data show two large-scale warming and shallowing-upward cycles, which are interpreted to be forced by the ~ 100 kyr eccentricity cycle of the Earth's orbit. Magnetostratigraphic data of the examined section reveal a long (113 m) reversed polarity interval, followed by a 7 m thick interval of normal polarity at the top. The inferred depositional rate of ~ 0.3 mm/yr, combined with biostratigraphic constraints by mollusks, suggests that the most logical correlation of the reversed interval is to chron C5Cr. This indicates that the Pag succession was deposited between 17.1 and 16.7 Ma and that it corresponds to the Burdigalian Stage of the Early Miocene, and the regional Karpatian Stage of the Central Paratethys. The high relative percentage of thermophilous pollen taxa, Engelhardia and Taxodium-type being the most prominent, generally indicates a subtropical humid climate for the SW Croatian part of the Dinaride Lake System. The observed warming trend is possibly related to the onset of the Miocene Climatic Optimum.     

Using chitosan-modified clays to control black-bloom-induced black suspended matter in Taihu Lake: Deposition and resuspension of black matter/clay flocs

A cyanobacteria-induced black bloom in Taihu Lake, 2007, subjected nearly one million Wuxi City residents to a drinking water crisis. This black bloom attracted wide attention in China and the rest of the world. However, black bloom is a highly weather-dependent event, and its rapid movement in lakes makes it difficult to predict where it will occur. Therefore, jar-tests and simulated flow experiments were performed to investigate the flocculation, deposition, and resuspension of black-bloom-induced black matter. At a dosage of 0.2 g L⁻¹ chitosan + 1 g L⁻¹ diatomite, 90% of the turbidity was removed within 1 h in jar-tests and in low flow speed simulated experiments. However, the black matter/clay flocs did not fully sink, and stratification of turbidity apparently occurred in the lower part of the tank during the simulated flow experiments. The resuspension experiments under simulated flow speeds showed that at a wind speed of 1–6 m s⁻¹ in Taihu Lake produced currents that did not greatly affect floc resuspension, but a wind speed of 3–4 m s⁻¹ produced waves that could induce floc resuspension. Quartz sand was sprayed over the flocs to verify its effect on inhibiting resuspension. A shear stress of 0.37 N m⁻² did not induce floc resuspension, which indicated that a wind speed of 6 m s⁻¹ would not induce floc resuspension in Taihu Lake.     

Global analysis of circulating metabolites in hibernating ground squirrels

Hibernation in mammals involves major alterations in nutrition and metabolism that would be expected to affect levels of circulating molecules. To gain insight into these changes we conducted a non-targeted LC–MS based metabolomic analysis of plasma using hibernating ground squirrels in late torpor (LT, T_b ~ 5 °C) or during an interbout arousal period (IBA, T_b ~ 5 °C) and non-hibernating squirrels in spring (T_b ~ 37 °C). Several metabolites varied and allowed differentiation between hibernators and spring squirrels, and between torpid and euthermic squirrels. Methionine and the short-chain carnitine esters of propionate and butyryate/isobutyrate were reduced in LT compared with the euthermic groups. Pantothenic acid and several lysophosphatidylcholines were elevated in LT relative to the euthermic groups, whereas lysophosphatidylethanolamines were elevated during IBA compared to LT and spring animals. Two regulatory lipids varied among the groups: sphingosine 1-phosphate was lower in LT vs. euthermic groups, whereas cholesterol sulfate was elevated in IBA compared to spring squirrels. Levels of long-chain fatty acids (LCFA) and total NEFA tended to be elevated in hibernators relative to spring squirrels. Three long-chain acylcarnitines were reduced in LT relative to IBA; free carnitine was also lower in LT vs. IBA. Our results identified several biochemical changes not previously observed in the seasonal hibernation cycle, including some that may provide insight into the metabolic limitations of mammalian torpor.     

Sap flow and water use in African baobab (Adansonia digitata L.) seedlings in response to drought stress

The African baobab (Adansonia digitata L.) is an important multi-purpose fruit tree with high potential for domestication in drier Africa. Although adult individuals are well-known to be drought resistant, only little has been reported on how young baobab trees can survive drought. Therefore, the aim of this study was to examine short-term soil drought effects on water relations of baobab seedlings. Baobab seedlings used a limited amount of stored water to buffer daily water deficits (~ 8.5 g d^− 1), which contributed up to only ~ 17.5% of daily water use and ~ 6% of total plant water. Under drought, a strong reduction in stomatal conductance (~ 85%) resulted in a midday leaf water potential of − 1 MPa and zero stem sap flow followed by significant leaf loss. Plant anatomy evidenced the presence of water storage tissues and the vulnerability to xylem embolism. The taproot was the most important plant part for water storage (68% of total plant water), suggesting root-succulence rather than stem-succulence. When drought intensified, limitation of leaf transpiration and/or root water uptake led to drought-enforced dormancy. Despite the large amounts of water stored in the taproot (~ 90%) and the stem (~ 75%), only a limited amount of stored water appeared to be used to sustain upper leaves and plant metabolism during the dormant period, and to facilitate recovery following water supply. Drought avoidance, conservative water use and the presence of internal stored water allow baobab seedlings to survive drought.     

Different roles of zinc plus arachidonic acid on insulin sensitivity between high fructose- and high fat-fed rats

AimsThis study was to determine the effects of zinc plus arachidonic acid (ZA) treatment on the insulin action in the specific ZA target organs using hyperinsulinemic euglycemic clamp method.Main methods18 Sprague–Dawley rats weighing ~ 130 g were divided into 3 groups of 6 rats and treated them with 1) normal rat chow, 2) high fructose (60.0%) diet only, or 3) the same fructose diet plus drinking water containing 10 mg zinc plus 50 mg arachidonic acid (AA)/L. In a separate study, male Wistar rats weighing ~ 250 g were fed normal rat chow (n = 4) or high fat (66.5%) diet with drinking water containing zero (n = 9) or 10 mg AA plus 20 mg zinc /L (n = 9). After 4 week treatment, insulin action was assessed using the hyperinsulinemic eguglycemic clamp technique.Key findingsHigh fructose feeding impaired suppression of hepatic glucose output by insulin compared to controls during the clamp procedure (4.39 vs. 2.35 mg/kg/min; p < 0.05). However, ZA treatment in high fructose-fed rats showed a significant improvement of hepatic insulin sensitivity compared to non-treatment controls (4.39 vs. 2.18 mg/kg/min; p < 0.05). Glucose infusion rates in Wistar rats maintained on a high fat diet (HFD) were significantly lower compared to control rats (22.8 ± 1.3 vs. 31.9 ± 1.4 mg/kg/min; p < 0.05). ZA treatment significantly improved (~ 43%) peripheral tissue insulin sensitivity in HFD fed animals (26.7 ± 1.3 [n = 9] vs. 22.8 ± 1.3 mg/kg/min; p < 0.05).SignificanceThese data demonstrate that ZA treatment is effective in improving glucose utilization in hyperglycemic rats receiving either a high-fructose or a high-fat diet.     

A new stage 3 millennial climatic variability record from a SW France speleothem

We present a new high resolution speleothem stable isotope record from the Villars Cave (SW-France) that covers part of marine isotope stage (MIS) 3. The Vil14 stalagmite grew between ~ 52 and 29 ka. The δ¹³C profile is used as a palaeoclimate proxy and clearly shows the interstadial substages 13, 12 and 11. The new results complement and corroborate previously published stalagmite records Vil9 and Vil27 from the same site. The Vil14 stalagmite chronology is based on 12 Th-U dating by MC-ICP-MS and 3 by TIMS. A correction for detrital contamination was done using the ²³⁰Th/²³²Th activity ratio measured on clay collected in Villars Cave. The Vil14 results reveal that the onset of Dansgaard–Oeschger (DO) events 13 and 12 occurred at ~ 49.8 ka and ~ 47.8 ka, respectively. Within uncertainties, this is coherent with the latest NorthGRIP time scale (GICC05-60 ka) and with speleothem records from Central Alps. Our data show an abrupt δ¹³C increase at the end of DO events 14 to 12 which coincides with a petrographical discontinuity probably due to a rapid cooling. As observed for Vil9 and Vil27, Vil14 growth significantly slowed down after ~ 42 ka and finally stopped ~ 29 ka ago where the δ¹³C increase suggests a strong climate deterioration that coincides with both North Atlantic sea level and sea surface temperature drop.     

Position—Specific contribution of interface tryptophans on membrane protein energetics

Interface tryptophans are key residues that facilitate the folding and stability of membrane proteins. Escherichia coli OmpX possesses two unique interface tryptophans, namely Trp76, which is present at the interface and is solvent-exposed, and Trp140, which is relatively more lipid solvated than Trp76 in symmetric lipid membranes. Here, we address the requirement for tryptophan and the consequences of aromatic amino acid substitutions on the folding and stability of OmpX. Using spectroscopic measurements of OmpX-Trp/Tyr/Phe mutants, we show that the specific mutation W76 → Y allows barrel assembly > 1.5-fold faster than native OmpX, and increases stability by ~ 0.4 kcal mol^− 1. In contrast, mutating W140 → F/Y lowers OmpX thermodynamic stability by ~ 0.4 kcal mol^− 1, without affecting the folding kinetics. We conclude that the stabilizing effect of tryptophan at the membrane interface can be position—and local environment—specific. We propose that the thermodynamic contributions for interface residues be interpreted with caution.     

CDP-choline-induced contractions in the mouse gastric fundus through purinoceptors and Rho/Rho-kinase signalling

AimsThis study aimed to investigate the effects of cytidine-5′-diphosphocholine (CDP-choline), an endogenous lipid precursor, on the reactivity of the mouse gastric fundus and to determine the mechanism(s) mediating its effects.Main methodsPossible contractile effect of CDP-choline (10^? 5–10^? 2 M) was investigated in the absence and presence of a muscarinic receptor antagonist, atropine (3 × 10^? 6 M), an acetylcholine esterase inhibitor, physostigmine (10^? 6 M), a Na⁺ channel blocker, tetrodotoxin (TTX, 3 × 10^? 6 M), a Rho-kinase inhibitor, Y-27632 (10^? 5 M), a purinoceptor antagonist, suramin (2 × 10^? 4 M), a nitric oxide synthase inhibitor, N^G-nitro-L-arginine (L-NA, 3 × 10^? 4 M), a Ca²⁺ channel blocker, nifedipine (10^? 6 M), an α₇ nicotinic receptor antagonist, methyllycaconitine citrate (MLA, 10^? 6 M) and a G protein (G_i/o) inhibitor, pertussis toxin (PTX, 2 μg/ml). The metabolites of CDP-choline, namely choline (10^? 4–10^? 2 M), cytidine 5′-triphosphate (CTP, 10^? 5–10^? 2 M), cytidine (10^? 5–10^? 2 M) and cytidine monophosphate (CMP, 10^? 3–10^? 2 M) were also tested. Besides, phosphorylation of MYPT1, which indicates Rho-kinase activity, was also detected.Key findingsCDP-choline produced contractions in a concentration-dependent manner. The contractions were not affected by atropine, physostigmine, TTX, PTX, MLA or L-NA. However, Y-27632, suramin or nifedipine partly reduced these contractions. CDP-choline increased phosphorylation of MYPT1. Among CDP-choline metabolites, cytidine had no contractile effects. However, choline induced considerable contractions, which were sensitive to atropine. CMP and CTP had also contractile activity, comparable to that of CDP-choline.SignificanceThese results suggest that CDP-choline produced contraction through, at least in part, purinoceptors and Rho/Rho-kinase signalling in the mouse gastric fundus.     

Inducible COX-2 dominates over COX-1 in prostacyclin biosynthesis: mechanisms of COX-2 inhibitor risk to heart disease

AimOur aim is to understand the molecular mechanisms of the selective nonsteroidal anti-inflammatory drugs (NSAID), cyclooxygenase-2 (COX-2) inhibitors', higher “priority” to reduce synthesis of the vascular protector, prostacyclin (PGI2), compared to that of nonselective NSAIDs.Main methodsCOX-1 or COX-2 was co-expressed with PGI2 synthase (PGIS) in COS-7 cells. The Km and initial velocity (½t Vmax) of the coupling reaction between COX-1 and COX-2 to PGIS were established. The experiment was further confirmed by a kinetics study using hybrid enzymes linking COX-1 or COX-2 to PGIS. Finally, COX-1 or COX-2 and PGIS were respectively fused to red (RFP) and cyanic (CFP) fluorescence proteins, and co-expressed in cells. The distances between COXs and PGIS were compared by FRET.Key findingsThe Km for converting arachidonic acid (AA) to PGI2 by COX-2 coupled to PGIS is ~ 2.0 μM; however, it was 3-fold more (~ 6.0 μM) for COX-1 coupled to PGIS. The Km and ½t Vmax for COX-2 linked to PGIS were ~ 2.0 μM and 20 s, respectively, which were 2–5 folds faster than that of COX-1 linked to PGIS. The FRET study found that the distance between COX-2-RFP and PGIS–CFP is shorter than that between COX-1-RFP and PGIS–CFP.SignificanceThe study provided strong evidence suggesting that the low Km, faster ½t Vmax, and closer distance are the basis for COX-2 dominance over COX-1 (coupled to PGIS) in PGI2 synthesis, and further demonstrated the mechanisms of selective COX-2 inhibitors with higher potential to reduce synthesis of the vascular protector, PGI2.     

Impact of the Middle Miocene climate transition on elongate,cylindrical foraminifera in the subtropical Pacific

Fifty-eight species of elongate, cylindrical benthic foraminifera (here referred to as the Extinction Group) belonging to genera that became extinct during the mid-Pleistocene Climate Transition (MPT), were documented (~ 50 kyr resolution) through the early middle Miocene (15–13 Ma) in two sites on opposite sides of the subtropical Pacific Ocean (ODP Sites 1146, South China Sea; ODP Site 1237, southeast Pacific). The study was undertaken to investigate the response of the Extinction Group (Ext. Gp) to the major cooling during the middle Miocene Climate Transition (MCT) to look for clues that might explain the causes of the extinction during the glacials of the mid-Pleistocene Climate Transition. Ext. Gp faunal differences between the two sites (attributed to regional and bathymetric differences in food supply to the seafloor) are greater than those that occurred through the 2 myr time span at either site. The middle Miocene Climate Transition was not an interval of enhanced species turnover or a decline in Ext. Gp abundance, in contrast to the major extinctions that occurred during the mid-Pleistocene Climate Transition. Distinct changes in the composition of the Ext. Gp faunas did occur through this time (more pronounced in Site 1237). At both sites the pre-middle Miocene Climate Transition faunas were transformed into their post-middle Miocene Climate Transition composition during the period of major cooling (14.0–13.7 Ma). During this transition interval the faunal composition swung back and forth between the two end member faunas. These faunal changes are attributed to changes in productivity (decrease in South China Sea, increase in southeast Pacific), brought about by major changes in global climate and continental aridity.     

Functional evaluation of tryptophans in glycolipid binding and membrane interaction by HET-C2, a fungal glycolipid transfer protein

HET-C2 is a fungal glycolipid transfer protein (GLTP) that uses an evolutionarily-modified GLTP-fold to achieve more focused transfer specificity for simple neutral glycosphingolipids than mammalian GLTPs. Only one of HET-C2's two Trp residues is topologically identical to the three Trp residues of mammalian GLTP. Here, we provide the first assessment of the functional roles of HET-C2 Trp residues in glycolipid binding and membrane interaction. Point mutants HET-C2^W208F, HET-C2^W208A and HET-C2^F149Y all retained > 90% activity and 80–90% intrinsic Trp fluorescence intensity; whereas HET-C2^F149A transfer activity decreased to ~ 55% but displayed ~ 120% intrinsic Trp emission intensity. Thus, neither W208 nor F149 is absolutely essential for activity and most Trp emission intensity (~ 85–90%) originates from Trp109. This conclusion was supported by HET-C2^W109Y/F149Y which displayed ~ 8% intrinsic Trp intensity and was nearly inactive. Incubation of the HET-C2 mutants with 1-palmitoyl-2-oleoyl-phosphatidylcholine vesicles containing different monoglycosylceramides or presented by lipid ethanol-injection decreased Trp fluorescence intensity and blue-shifted the Trp λ_max by differing amounts compared to wtHET-C2. With HET-C2 mutants for Trp208, the emission intensity decreases (~ 30–40%) and λ_max blue-shifts (~ 12 nm) were more dramatic than for wtHET-C2 or F149 mutants and closely resembled human GLTP. When Trp109 was mutated, the glycolipid induced changes in HET-C2 emission intensity and λ_max blue-shift were nearly nonexistent. Our findings indicate that the HET-C2 Trp λ_max blue-shift is diagnostic for glycolipid binding; whereas the emission intensity decrease reflects higher environmental polarity encountered upon nonspecific interaction with phosphocholine headgroups comprising the membrane interface and specific interaction with the hydrated glycolipid sugar.     

Structural changes that occur upon photolysis of the Fe(II)a3–CO complex in the cytochrome ba3-oxidase of Thermus thermophilus: A combined X-ray crystallographic and infrared spectral study demonstrates CO binding to CuB

The purpose of the work was to provide a crystallographic demonstration of the venerable idea that CO photolyzed from ferrous heme-a₃ moves to the nearby cuprous ion in the cytochrome c oxidases. Crystal structures of CO-bound cytochrome ba₃-oxidase from Thermus thermophilus, determined at ~ 2.8–3.2 Å resolution, reveal a Fe–C distance of ~ 2.0 Å, a Cu–O distance of 2.4 Å and a Fe–C–O angle of ~ 126°. Upon photodissociation at 100 K, X-ray structures indicate loss of Fe_a3–CO and appearance of Cu_B–CO having a Cu–C distance of ~ 1.9 Å and an O–Fe distance of ~ 2.3 Å. Absolute FTIR spectra recorded from single crystals of reduced ba₃–CO that had not been exposed to X-ray radiation, showed several peaks around 1975 cm^? 1; after photolysis at 100 K, the absolute FTIR spectra also showed a significant peak at 2050 cm^? 1. Analysis of the ‘light’ minus ‘dark’ difference spectra showed four very sharp CO stretching bands at 1970 cm^? 1, 1977 cm^? 1, 1981 cm^? 1, and 1985 cm^? 1, previously assigned to the Fe_a3–CO complex, and a significantly broader CO stretching band centered at ~ 2050 cm^? 1, previously assigned to the CO stretching frequency of Cu_B bound CO. As expected for light propagating along the tetragonal axis of the P4₃2₁2 space group, the single crystal spectra exhibit negligible dichroism. Absolute FTIR spectrometry of a CO-laden ba₃ crystal, exposed to an amount of X-ray radiation required to obtain structural data sets before FTIR characterization, showed a significant signal due to photogenerated CO₂ at 2337 cm^? 1 and one from traces of CO at 2133 cm^? 1; while bands associated with CO bound to either Fe_a3 or to Cu_B in “light” minus “dark” FTIR difference spectra shifted and broadened in response to X-ray exposure. In spite of considerable radiation damage to the crystals, both X-ray analysis at 2.8 and 3.2 Å and FTIR spectra support the long-held position that photolysis of Fe_a3–CO in cytochrome c oxidases leads to significant trapping of the CO on the Cu_B atom; Fe_a3 and Cu_B ligation, at the resolutions reported here, are otherwise unaltered. This article is part of a Special Issue entitled: Respiratory Oxidases.     

Sulforaphane improves dysregulated metabolic profile and inhibits leptin-induced VSMC proliferation: Implications toward suppression of neointima formation after arterial injury in western diet-fed obese mice

Sulforaphane (SFN), a dietary phase-2 enzyme inducer that mitigates cellular oxidative stress through nuclear factor erythroid 2-related factor 2 (Nrf2) activation, is known to exhibit beneficial effects in the vessel wall. For instance, it inhibits vascular smooth muscle cell (VSMC) proliferation, a major event in atherosclerosis and restenosis after angioplasty. In particular, SFN attenuates the mitogenic and pro-inflammatory actions of platelet-derived growth factor (PDGF) and tumor necrosis factor-α (TNFα), respectively, in VSMCs. Nevertheless, the vasoprotective role of SFN has not been examined in the setting of obesity characterized by hyperleptinemia and insulin resistance. Using the mouse model of western diet-induced obesity, the present study demonstrates for the first time that subcutaneous delivery of SFN (0.5 mg/Kg/day) for ~ 3 weeks significantly attenuates neointima formation in the injured femoral artery [↓ (decrease) neointima/media ratio by ~ 60%; n = 5–8]. This was associated with significant improvements in metabolic parameters, including ↓ weight gain by ~ 52%, ↓ plasma leptin by ~ 42%, ↓ plasma insulin by ~ 63%, insulin resistance [↓ homeostasis model assessment of insulin resistance (HOMA-IR) index by ~ 73%], glucose tolerance (↓ AUC_GTT by ~ 24%), and plasma lipid profile (e.g., ↓ triglycerides). Under in vitro conditions, SFN significantly decreased leptin-induced VSMC proliferation by ~ 23% (n = 5) with associated diminutions in leptin-induced cyclin D1 expression and the phosphorylation of p70S6kinase and ribosomal S6 protein (n = 3–4). The present findings reveal that, in addition to improving systemic metabolic parameters, SFN inhibits leptin-induced VSMC proliferative signaling that may contribute in part to the suppression of injury-induced neointima formation in diet-induced obesity.