Cardenolides from Asclepias asperula subsp. Capricornu and A. Viridis

6′-O-(E-4-hydroxycinnamoyl) Desglucouzarin, the first cardenolide containing a cinnamoyl ester moiety, has been isolated from the ethanolic extract of the milkweed, Asclepias asperula. In addition, five known cardenolides were isolated and identified from A. asperula and A. viridis.     

Cytotoxicity of cardenolides and cardenolide glycosides from Asclepias curassavica

A new cardenolide, 12β,14β-dihydroxy-3β,19-epoxy-3α-methoxy-5α-card-20(22)-enolide (6), and a new doubly linked cardenolide glycoside, 12β-hydroxycalotropin (13), together with eleven known compounds, coroglaucigenin (1), 12β-hydroxycoroglaucigenin (2), calotropagenin (3), desglucouzarin (4), 6′-O-feruloyl-desglucouzarin (5), calotropin (7), uscharidin (8), asclepin (9), 16α-hydroxyasclepin (10), 16α-acetoxycalotropin (11), and 16α-acetoxyasclepin (12), were isolated from the aerial part of ornamental milkweed, Asclepias curassavica and chemically elucidated through spectral analyses. All the isolates were evaluated for their cytotoxic activity against HepG2 and Raji cell lines. The results showed that asclepin (9) had the strongest cytotoxic activity with an IC₅₀ value of 0.02 μM against the two cancer cell lines and the new compound 13 had significant cytotoxic activity with IC₅₀ values of 0.69 and 1.46 μM, respectively.     

Effects of the MyoG Gene on the Partial Growth Traits in Pigs

Single-nucleotide polymorphisms of the MyoG gene were tested using PCR-SSCP in different pig breeds including Landrace, Large White, Duroc, Shanxi Black, and Mashen pigs, and the effects of the MyoG gene on the birth weight, the weaning weight, the 6-month body weight, and the backfat thickness were also analyzed. On the basis of the published sequence of the porcine MyoG gene, ten pairs of primers were designed, and one polymorphism was found in the PCR product amplified with In2-3 primers. The results showed that: (1) the Landrace, the Large White, and the Duroc breeds differ significantly (P < 0.05) in genotype distribution from the Shanxi Black and the Mashen breeds; (2) On the basis of the fixed effect model, significant differences were found in the birth weight and the backfat thickness among the different MyoG genotypes, whereas no significant differences existed in the weaning weight and the 6-month body weight; (3) Using least square analysis, it was seen that individuals of the BB genotype had significantly less (P < 0.01) birth weight than those of the AA and AB genotypes, with the order being AA>AB>BB; the pigs of the AA genotype had significantly lower (P < 0.01) backfat thickness than those of the AB and BB genotypes, with the order being AA<AB<BB. These results suggest that the genotype has significant effects on the individual birth weight and the backfat thickness, and that the selection of the A allele is favored with regard to the birth weight and the backfat thickness.     

Valbro : un nouveau site à vertébrés de l’Oligocène inférieur (MP22) de France (Quercy). I – Contexte géologique ; Mammalia : Rodentia,Hyaenodontida, Carnivora

The locality Valbro (Quercy, France) has yielded a rich fossil vertebrate fauna. Here are presented the geological context, a preliminary faunal list and the systematics of the Rodentia and carnivores, the most abundant taxa of the fauna. The Rodentia are known from 11 taxa: the Theridomyidae Blainvillimys gregarius Schlosser, 1884, Blainvillimys ?gemellus Vianey-Liaud, 1989, Issiodoromys medius (Vianey-Liaud, 1976) and S. cayluxi Schlosser, 1884, the Aplodontidae Plesispermophilus angustidens Filhol, 1882, the Gliridae Butseloglis tenuis (Bahlo, 1975), Butseloglis micio (Misonne, 1957) and Bransatoglis planus (Bahlo, 1975), the Sciuridae Palaeosciurus goti Vianey-Liaud, 1974 and cf. Oligopetes sp., and the Cricetidae Atavocricetodon sp. aff. A. nanus (Pelaez-Campomanes, 1995). Thirteen taxa of carnivores are present at Valbro; among the Hyaenodontida: Hyaenodon leptorhynchus Laizer et Parieu, 1838 and cf. Apterodon sp.; among the Carnivora: a Feliformia gen. et sp. indet., the Nimravidae Nimravus intermedius (Filhol, 1872), Dinailurictis bonali Helbing, 1922 and Nimravidae gen. et sp. indet. (Certainly belonging to one the two already identified species), the Ursida Pachycynodon crassirostris Schlosser, 1888, Pachycynodon sp., Amphicynodon sp. 1 cf. A. typicus (Schlosser, 1888), Amphicynodon ? sp. 2 and Ursida gen. et sp. indet. cf. Pachycynodon boriei (Filhol, 1876), the Mustelida M. olivieri Bonis, 1997, the Carnivora incertae sedis Palaeogale sectoria (Gervais, 1848–1852), the Arctoidea gen. et sp. indet. and Carnivora gen. et sp. indet. (probably representing taxa already identified at lower taxonomic levels). The evolutionary grade of B. gregarius of Valbro compares to that of the species from Mas de Got, La Plante 2 and Cavalé, which supports an MP22 age for Valbro. This datation is further supported by the association of Theridomyinae yielded by the locality (B. gregarius, B. gemellus, I. medius and S. cayluxi), and by the presence of a N. intermedius that well compares the material of the species from Villebramar, La Plante et Mas de Got, of D. bonali, P. sectoria, M. olivieri (species known only from Mas de Got), and by additional evidence from the remaining vertebrates from Valbro, especially the squamates. Despite a limited amount of specimens (341 specimens have been studied), the faunas of rodents and carnivores from Valbro are the most speciose and diverse known for the MP22 level and, for the Carnivora fauna in particular, for the Oligocene of the Old World.     

全球陆地生态系统光合作用与呼吸作用的温度敏感性

基于全球647套通量数据,定量分析了全球尺度下生态系统光合作用和呼吸作用的温度敏感性(Q10)随纬度、气候和植被的分布规律。结果表明:在全球尺度下,光合作用和呼吸过程的温度敏感性(Q10,G和Q10,R)都随纬度的升高而增加,其中Q10,G和Q10,R的均值分别为3.99±0.21和2.28±0.074。除热带多树草原、常绿落叶林外,Q10,G均大于Q10,R值。不同植被类型的温度敏感性存在显著性差异,表现为:针叶林阔叶林;落叶林常绿林,其中生态系统的季节性变异是造成差异的主要原因。当植被类型和纬度区域共同影响Q10值时,植被类型对Q10值的总变异贡献更大。气候类型对Q10,G和Q10,R都有显著影响。在气候带上,干旱带的Q10,G最小,而冷温带的Q10,G最高。不同气候类型下(除温带草原气候外)的Q10,G都大于Q10,R。在极端条件下,温度可能不在是主导因素,而水分对温度敏感性的影响不可忽略,今后的研究需要更多的关注生态系统温度敏感性对水分变化的响应。     

Differences in resource storage pattern between Laminaria longissima and Laminaria diabolica (Laminariaceae; Phaeophyta) reflecting their morphological characteristics

The kelps Laminaria longissima and L. diabolica, belonging to the groups of L. angustata and L. japonica, respectively, differ greatly in their morphological characteristics although their geographical distributions overlap widely along the eastern coast of Hokkaido. To clarify the interaction between the morphological and physiological characteristics of the two species, and their link with environmental variables, hatchery-raised young sporophytes of L. longissima and L. diabolica collected from Hokkaido were cultivated simultaneously under similar conditions in Matsushima Bay, Miyagi, from January to July 2004. Seasonal morphological characteristics, gross photosynthetic rate, nutrient uptake rates, and resource contents were examined. The blade lengths of L. longissima and L. diabolica reached a maximum of 329.9 cm and 256.7 cm, respectively, in April to May, and decreased to 284.4 cm and 68.6 cm, respectively, in July. The total elongation length of L. longissima (412.5 cm) was similar to that of L. diabolica (373.8 cm). However, the total erosion length of L. longissima (145.9 cm) was approximately half that of L. diabolica (302.9 cm). The gross photosynthetic rate and uptake rates of NH₄-N, NO₃-N, and PO₄-P of the two species were similar. However, the carbon, nitrogen, and phosphorus contents were transferred and stored in the whole blade tissues in the case of L. longissima, but in the meristem of L. diabolica from May to June. These results suggest that morphological differences are a response to different resource storage patterns. The storage patterns of L. longissima and L. diabolica are likely to be genetically fixed characteristics, which have evolved in adaptation to the specific habitat environments of the groups of L. angustata and L. japonica. The low water temperature and rich nutrients provided by the Oyashio Current are conducive to storage of resources in the whole blade tissues and a large surface area retained for photosynthesis and nutrient uptake in the L. angustata group. Conversely, high temperature and poor nutrients, or large fluctuations in these parameters, provided by the Tsushima Warm Current are more conducive to intensive storage of resources in the meristem for maturation and further growth in the L. japonica group. L. diabolica retains the storage pattern of the L. japonica group but grows in regions affected by the Oyashio Current, allowing it to become the widest distributed Laminaria species.     

Middle triassic Parapopanoceratidae and Nathorstitidae (ammonoidea) of Boreal region: Internal structure,ontogeny and phylogenetic patterns

Internal structure of Stenopopanoceras mirabilePopow, Parapopanoceras sp., P. mediumMcLearn, P. janaense (Popow), P. paniculatumPopow, P. asseretoiDagys & Ermakova, Indigirites krugiPopow, Nathorstites mcconnelli (Whiteaves), N. gibbosusStolley, N. lenticularis (Whiteaves), N. argatassensis (Popow) is described and also ontogenetic development of species I. krugiPopow and N. mcconnelli (Whiteaves) originating from middle triassic deposits of northern part of Middle Siberia, north-eastern part of the USSR and Svalbard is given. For the species mentioned above data is given on the position of siphuncle, shape and dimensions of protoconch, caecum, ammonitella, the angle of primary varix, structure of septal necks and the nature of their change in the course of ontogeny. Significance of these signs for taxonomy and phylogeny of triassic ammonoidea is analysed. Intergeneric difference of Stenopopanoceras and Parapopanoceras is established according to the position of siphuncle on the first whorl of the shell. On the basis of the analysis of internal structure and ontogenetic development of suture line and morphological signs Indigirites is considered to belong to the family Hungaritidae (subfamily Longobarditinae). Phylogenetic patterns of the ammonoidea species described are discussed.     

Two new species of Dunkleosteus Lehman, 1956, from the Ohio Shale Formation (USA,Famennian) and the Kettle Point Formation (Canada,Upper Devonian), and a cladistic analysis of the Eubrachythoraci (Placodermi,Arthrodira)

Two pachyosteomorph arthrodires are described from the Upper Devonian of North America, Dunkleosteus raveri sp. nov. and Dunkleosteus amblyodoratus sp. nov. (from the Appalachian and the Michigan basins, respectively). Dunkleosteus raveri sp. nov. is found stratigraphically below the only other species within the Ohio Shale Formation (Famennian), and is characterized by the retention of a primitive dermal ornamentation, and shares with Dunkleosteus terrelli ( Newberry, 1873 ) the possession of articular facets on the parasphenoid. Dunkleosteus amblyodoratus sp. nov. (Kettle Point Formation, Upper Devonian), known only from incomplete nuchal, paranuchal, and parasphenoid plates, also retains dermal tuberculation, and is characterized by tapering articular facets on the parasphenoid. The current study reviews three North American species of Dunkleosteus (Du. terrelli, Du. raveri sp. nov. , and Du. amblyodoratus sp. nov. ). Formerly, two subgroups were recognized within the Dinichthyidae, the terrelli and herzeri groups, which are shown here to be members of distinct clades. Dinichthys herzeri Newberry, 1868 (the namesake member of the herzeri group) is now the sole member of the family Dinichthyidae comb. nov. , and is assigned to the Aspinothoracidi sensu Miles & Dennis, 1979 . Heintzichthys Whitley, 1933 and Gorgonichthys Claypole, 1892 (additional members of the herzeri group) were previously placed among the aspinothoracid arthrodires. Likewise, Holdenius holdeni Dunkle & Bungart, 1942 (the final member of the herzeri group) and Hadrosteus rapax Gross, 1932 are now placed within the Aspinothoracidi. The remaining taxa, formerly assigned to the Dinichthyidae (i.e. Dunkleosteus Lehman, 1956 ; Eastmanosteus Obruchev, 1964 ; Golshanichthys Lelièvre, Janvier, & Goujet, 1981 ; and Westralichthys Long, 1987 ), are united within the Dunkleosteidae Stensiö, 1963 . © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159 , 195–222.     

Litter Decomposition in Temperate Peatland Ecosystems: The Effect of Substrate and Site

Abstract The large accumulation of organic matter in peatlands is primarily caused by slow rates of litter decomposition. We determined rates of decomposition of major peat-forming litters of vascular plants and mosses at five sites: a poor fen in New Hampshire and a bog hummock, a poor fen, a beaver pond margin and a beaver pond in Ontario. We used the litterbag technique, retrieving triplicate litterbags six or seven times over 3–5 years, and found that simple exponential decay and continuous-quality non-linear regression models could adequately characterize the decomposition in most cases. Within each site, the rate of decomposition at the surface was generally Typha latifolia leaves = Chamaedaphne calyculata leaves = Carex leaves > Chamaedaphne calyculata stems > hummock Sphagnum = lawn/hollow Sphagnum, with exponential decay constant (k) values generally ranging from 0.05 to 0.37 and continuous-quality model initial quality (q ₀ ) values ranging from 1.0 (arbitrarily set for Typha leaves) to 0.7 (Sphagnum). In general, surface decay rates were slowest at the bog hummock site, which had the lowest water table, and in the beaver pond, which was inundated, and fastest at the fens. The continuous-quality model site decomposition parameter (u ₀ ) ranged from 0.80 to 0.17. Analysis of original litter samples for carbon, nitrogen and proximate fractions revealed a relatively poor explanation of decomposition rates, as defined by k and q ₀ , compared to most well-drained ecosystems. Three litters, roots of sedge and a shrub and Typha leaves, were placed at depths of 10, 30 and 60 cm at the sites. Decomposition rates decreased with depth at each site, with k means of 0.15, 0.08 and 0.05 y⁻¹ at 10, 30 and 60 cm, respectively, and u ₀ of 0.25, 0.13 and 0.07. These differences are primarily related to the position of the water table at each site and to a lesser extent the cooler temperatures in the lower layers of the peat. The distinction between bog and fen was less important than the position of the water table. These results show that we can characterize decomposition rates of surface litter in northern peatlands, but given the large primary productivity below-ground in these ecosystems, and the differential rates of decomposition with depth, subsurface input and decomposition of organic matter is an important and relatively uncertain attribute.     

北京典型树种木质组织碳释放速率温度敏感性的时间变化规律和铅锤分异特征

采用红外气体分析法(IRGA)于2014年1—12月原位测定了北京市4个典型树种(国槐Sophora japonica,旱柳Salix matsudana,华北落叶松Larix principis-rupprechtii和侧柏Platycladus orientalis)在不同高度上的木质组织CO_2通量速率(E_(CO_2)),旨在比较不同树种间E_(CO_2)及其温度敏感性(Q_(10))的时间变化规律和铅锤分异特征。研究结果显示:(1)4个树种的E_(CO_2)均表现为单峰型季节变化规律,生长月份内的E_(CO_2)显著高于非生长月份,温度和枝干的径向生长是影响E_(CO_2)季节变化的主要因素;(2)E_(CO_2)对温度的敏感性在夏季月份明显降低,且出现明显的垂直分异:Q_(10)随测量高度的增加而增加,呈现出非连续的阶梯分布;(3)在日间尺度上,阔叶树种E_(CO_2)对温度的感性系数Q_(10)出现昼夜不对称现象,晚上Q_(10)明显升高。准确量化E_(CO_2)的时间变化规律和铅锤分异特征,细化不同时间尺度下E_(CO_2)对温度的响应特征,成为准确估算木质组织碳排放的前提条件。     

Acid glycosidases from hen oviduct and egg albumen

Activities of seven acid glycosidases: β-N-acetylhexosaminidase (β-HEX), α- and β-galactosidase (α- and β-GAL), α- and β-mannosidase (α- and β-MAN), α-glucosidase and α-fucosidase in magnum region of hen (Gallus gallus domesticus) oviduct, and four acid glycosidases: β-HEX, β-GAL, α- and β-MAN in egg albumen, were investigated. β-HEX from magnum and egg albumen hydrolysed 4-methylumbelliferyl-β-N-acetylhexosamine-6-sulphate (4-MeUmbGlcNAc-6-SO₄) like mammalian β-HEX form A. Multiple forms of magnum and egg albumen β-HEX, β-GAL, α- and β-MAN were separated by strong anion exchange chromatography and chromatofocusing method. Chromatofocusing of the magnum resulted in the appearance of multiple forms for β-HEX with pI of 6.18, 5.43, 5.55, 5.34, 5.27 and 5.16, for β-GAL with pI of 4.98, 4.84, 4.77, 4.64 and 4.68–4.63, for α-MAN with pI of ≥ 7.4, 6.75, 6.62 and 6.26, and for β-MAN two forms with pI of 6.37 and 5.77. Chromatofocusing of egg albumen yields multiple forms for β-HEX with pI of 6.24, 6.08, 5.55 and 5.35, for β-GAL two forms with pI of 5.10 and 4.86–4.80 for α-MAN multiple forms with pI of ≥ 7.4, 6.80, 6.60 and 6.30, and for β-MAN forms with pI of 6.30 and 5.77. In conclusion, this study was the first to show β-HEX activity against 4-MeUmbGlcNAc-6-SO₄ in the magnum and albumen of bird eggs, corresponding to β-HEX A activity in mammals. Main multiple forms of β-HEX, β-GAL, α- and β-MAN occurring in the magnum were revealed in the egg albumen. Comparison with a cock of the same breed showed that hen egg magnum and albumen has the same multiple forms of the enzymes that are found in the epididymides and seminal plasma of the cock.     

Influence of mixed antibiotics on Microcystis aeruginosa during the application of glyphosate and hydrogen peroxide algaecides

Antibiotics regulate various physiological functions in cyanobacteria and may interfere with the control of cyanobacterial blooms during the application of algaecides. In this study, Microcystis aeruginosa was exposed to H₂O₂ and glyphosate for 7 d in the presence of coexisting mixed antibiotics (amoxicillin, spiramycin, tetracycline, ciprofloxacin, and sulfamethoxazole) at an environmentally relevant concentration of 100 ng · L^?1. The mixed antibiotics significantly (P < 0.05) alleviated the growth inhibition effect of 15–45 μM H₂O₂ and 40–60 mg · L^?1 glyphosate. According to the increased contents of chlorophyll a and protein, decreased content of malondialdehyde, and decreased activities of superoxide dismutase and glutathione S‐transferase, antibiotics may reduce the toxicity of the two algaecides through the stimulation of photosynthesis and the reduction in oxidative stress. The presence of coexisting antibiotics stimulated the production and release of microcystins in the M. aeruginosa exposed to low concentrations of algaecides and posed an increased threat to aquatic environments. To eliminate the secondary pollution caused by microcystins, high algaecide doses that are ≥45 μM for H₂O₂ and ≥60 mg · L^?1 for glyphosate are recommended. This study provides insights into the ecological hazards of antibiotic contaminants and the best management practices for cyanobacterial removal under combined antibiotic pollution conditions.     

Phylogenetic relationships within the South American fish family Anostomidae (Teleostei,Ostariophysi, Characiformes)

Analysis of a morphological dataset containing 152 parsimony‐informative characters yielded the first phylogenetic reconstruction spanning the South American characiform family Anostomidae. The reconstruction included 46 ingroup species representing all anostomid genera and subgenera. Outgroup comparisons included members of the sister group to the Anostomidae (the Chilodontidae) as well as members of the families Curimatidae, Characidae, Citharinidae, Distichodontidae, Hemiodontidae, Parodontidae and Prochilodontidae. The results supported a clade containing Anostomus, Gnathodolus, Pseudanos, Sartor and Synaptolaemus (the subfamily Anostominae sensu Winterbottom) albeit with a somewhat different set of relationships among the species within these genera. Anostomus as previously recognized was found to be paraphyletic and is split herein into two monophyletic components, a restricted Anostomus and the new genus Petulanos gen. nov. , described herein. Laemolyta appeared as sister to the clade containing Anostomus, Gnathodolus, Petulanos, Pseudanos, Sartor and Synaptolaemus. Rhytiodus and Schizodon together formed a well‐supported clade that was, in turn, sister to the clade containing Anostomus, Gnathodolus, Laemolyta, Petulanos, Pseudanos, Sartor and Synaptolaemus. Anostomoides was sister to the clade formed by these nine genera. Leporinus as currently defined was not found to be monophyletic, although certain clades within that genus were supported, including the species with subterminal mouths in the former subgenus Hypomasticus which we recognize herein as a genus. Abramites nested in Leporinus, and Leporellus was found to be the most basal anostomid genus. The presence of cis‐ and trans‐Andean species in Abramites, Leporellus, Leporinus and Schizodon, all relatively basal genera, suggests that much of the diversification of anostomid species pre‐dates the uplift of the Andean Cordilleras circa 11.8 million years ago. Several important morphological shifts in anostomid evolution are illustrated and discussed, including instances of convergence and reversal. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society, 2008, 154 , 70–210.     

Characterization of Inhibitor-Sensitive and -Resistant Adenosine Transporters in Cultured Human Fetal Astrocytes

Abstract: The kinetic characteristics of [³H]adenosine uptake, the extent to which accumulated [³H]adenosine was metabolized, the effects such metabolism had on measurements of apparent Michaelis-Menten kinetic values of K_T and V_max, and the sensitivities with which nucleoside transport inhibitors blocked [³H]adenosine accumulations were determined in cultured human fetal astrocytes. K_T and V_max values for accumulations of [³H]-labeled purines using 15-s incubations in the absence of the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) and the adenosine kinase inhibitor 5′-iodotubercidin (ITU) were 6.2 µM and 0.15 nmol/min/mg of protein for the high-affinity and 2.6 mM and 21 nmol/min/mg of protein for the low-affinity components respectively. In the presence of EHNA and ITU, where <4% of accumulated [³H]adenosine was metabolized, transport per se was measured, and kinetic values for K_T and V_max were 179 µM and 5.2 nmol/min/mg of protein, respectively. In the absence of EHNA and ITU, accumulated [³H]adenosine was rapidly metabolized to AMP, ADP, and ATP, and caused an appearance of “concentrative” uptake in that the intracellular levels of [³H]-labeled purines (adenosine plus its metabolites) were 1.4-fold higher than in the medium. No apparent concentrative accumulations of [³H]adenosine were found when assays were conducted using short incubation times in the absence or presence of EHNA and ITU. The nucleoside transport inhibitors dipyridamole (DPR), nitrobenzylthioinosine (NBI), and dilazep biphasically inhibited [³H]adenosine transport; for the inhibitor-sensitive components the IC₅₀ values were 0.7 nM for NBI, 1.3 nM for DPR, and 3.3 nM for dilazep, and for the inhibitor-resistant component the IC₅₀ values were 2.5 µM for NBI, 5.1 µM for dilazep, and 39.0 µM for DPR. These findings, in cultured human fetal astrocytes, represent the first demonstration of inhibitor-sensitive and -resistant adenosine transporters in nontransformed human cells.     

Nouvelles données sur le genre Imerites Rouchadzé, 1933 (Ammonitida,Ancyloceratina) ; réponses et précisions apportées au travail de Bert,Delanoy et Canut, 2009

This work is mainly an answer to the work of Bert, Delanoy and Canut, on the genus Imerites Rouchadzé, and we add also some points to the knowledge of this genus. An answer is given for the implication of one of the authors (J.V.) of this present work, about the origin of the genus Imerites Rouchadzé, and some significant bibliographic references, not named in the 2009 work, are specified and placed in their context. The characteristics of the ornamental stage “Pseudoshasticrioceras” mainly defined by an oval and compressed section of the whorls and by regular ventrolateral clavi, well marked, which frame the venter in a typical way (Bert et al., p. 181), don’t allow to affirm the presence of this stage during the ontogeny of all species of the genus Imerites Rouchadzé, contrary to what Bert, Delanoy and Canut (p. 30, 31) wrote. The type-species of the genus Imerites Rouchadzé is made clear by the application of the rules of the I.C.Z.N. code. The article 23, with the paragraph 23.1 of the I.C.Z.N. code, imply the senior synonymy of Imerites cristatus (Orbigny) on Imerites giraudi (Kilian), even if the Kilian's species is still the type species of the genus Imerites Rouchadzé. The taxinomical validity of the species Imerites cristatus (Orbigny) is confirmed. Its morphological variability is clarified by the distinction of four referential morphotypes, Cristatus, Giraudi, Favrei and Raricostatum. Consequently, the assumption of a shape-dimensional dimorphism into the populations of the genus Imerites Rouchadzé is rejected, awaiting irrefutable proof. A new species, Imerites stephaniae sp. nov., is described. It was collected in the Gerhardtia sartousiana Zone, in the uppermost part of the Gerhardtia provincialis Subzone, and it is a probable ancestor of the cogeneric species of the Imerites giraudi Zone.     

Arecoline and the 30–100 kDa fraction of areca nut extract differentially regulate mTOR and respectively induce apoptosis and autophagy: a pilot study

Areca nut (AN) is recognized as a human carcinogen; however, few studies of the cytotoxic effects of AN ingredients on cells have been reported. In Taiwan, AN, lime and inflorescence of Piper betle are the common components of betel quid (BQ). We recently noticed that extract of AN (ANE), but not those of lime and inflorescence of Piper betle, induces rounding cell morphology and nuclear shrinkage in different types of carcinoma cells. In this study, the rounding cell activity was first traced to the partially purified ≥10 kDa fraction (ANE ≥ 10 K) and subsequently to the 30–100 kDa fraction (ANE 30–100 K). ANE and ANE ≥10 K stimulated nuclear shrinkage (P < 0.001 in both cases) and the clearance of the cytoplasm. ANE, ANE ≥ 10 K, and ANE 30–100 K induced the cleavage of LC3-I (P < 0.05, 0.01, and 0.05, respectively) and the emergence of autophagic vacuoles (AVs) and acidic vesicles. On the other hand, arecoline (Are, the major alkaloid of AN) triggered caspase-3 activation, peri-nuclear chromatin condensation, and micronucleation. Meanwhile, ANE 30–100 K, but not Are, inhibited the phosphorylation of the mammalian target of rapamycin (mTOR)-Ser²⁴⁴⁸. In conclusion, this study demonstrates that different AN ingredients exerting differential impact on mTOR-Ser²⁴⁴⁸ phosphorylation are capable of triggering apoptosis and autophagy. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Shyun-Yeu Liu, Mei-Huei Lin, Yu-Rung Hsu and Ya-Yun Shih contributed equally to this work.     

Cold activity and tolerance of the entomopathogenic fungus Tolypocladium spp. to UV-B irradiation and heat

Studies on the stress resistance of insect-pathogenic fungi are very important to better understand the survival of these organisms in the environment. In this study, we examined the cold activity (8 ± 1 °C for 7 days), UV-B tolerance (Quaite-weighted UV-B irradiance at 847.90 mW m⁻² for 1, 2, 3, and 4 h), and wet-heat tolerance (45 °C for 1, 2, 3, and 4 h) of two isolates of Tolypocladiumcylindrosporum (ARSEF 3392 and 5558), one isolate of Tolypocladium geodes (ARSEF 3275), and two isolates of Tolypocladium inflatum (ARSEF 4772 and 4877) based on their germination, compared with Metarhizium robertsii (ARSEF 2575). After 3 h of UV-B exposure, T. cylindrosporum germinated at a greater rate than the other Tolypocladium species and had similar viability to that of the M. robertsii. Most Tolypocladium isolates, however, were less UV-B tolerant than M. robertsii. The T.cylindrosporum isolates were also the most thermotolerant, with similar tolerance to the M. robertsii. The isolates of T. inflatum and T. geodes, which had similar heat tolerance, were the least heat tolerant compared with the isolates of T. cylindrosporum and M. robertsii. After 4 h of heat exposure, the germination of T. inflatum and T. geodes isolates was not significantly different. For cold activity, both T.cylindrosporum isolates germinated to ca. 100% in only 3 days. Approximately 50% of the two T. inflatum isolates germinated, and less than 5% of T. geodes germinated after 3 days. All fungal isolates, however, completely germinated by the seventh day, except M.robertsii. The isolates of T. cylindrosporum, therefore, were the most heat and UV-B tolerant, and had the highest cold activity compared to the other species. The tolerance of M. robertsii to UV-B radiation and heat was similar to that of T.cylindrosporum.     

Mexican Arnica Anti–Inflammatory Action: Plant Age is Correlated with the Concentration of Anti–Inflammatory Sesquiterpenes in the Medicinal Plant <Emphasis Type="Italic">Heterotheca inuloides</Emphasis> Cass. (Asteraceae)<Superscript>1</Superscript>

Mexican Arnica Anti–Inflammatory Action: Plant Age Is Correlated with the Concentration of Anti–inflammatory Sesquiterpenes in the Medicinal Plant Heterotheca inuloides Cass. (Asteraceae). Mexican árnica (Heterotheca inuloides Cass.) is a widely used anti–inflammatory medicinal plant in Mexican folk medicine. Although it has been suggested that plant age, fertilization, and harvesting regime influence the concentration of secondary compounds affecting the therapeutic activity of the plant, the effect of these variables on the concentration of the Mexican árnica anti–inflammatory compounds was not known. We quantified anti–inflammatory sesquiterpenes (caryolan–1, 9β–diol, cadalen–15–oic acid, 7–hydroxycadalene, 4–hydroxy–2–isopropyl–4, 7–dimethyl–1[4H] naftalinone, 7–hydroxy–4αH–3, 4–dihydrocadalene, β–caryophyllene, and β–caryophyllene epoxide) in Mexican árnica plants subjected to fertilization and successive harvests of flowering stems, conditions that mimic the cultivation and harvesting for árnica in México. Fertilization and successive harvesting and their interaction had no significant effect on the concentration of anti–inflammatory compounds. However, the concentrations of these compounds were 60% higher in flowering stems from 15–month–old plants than in those from 4– or 8–month–old plants and was independent of the number of harvests and fertilization regime applied.     

Hagfish phylogeny and taxonomy,with description of the new genus Rubicundus (Craniata,Myxinidae)

A recent phylogenetic analysis of the Myxinidae based on the 16S rRNA gene resulted in synonymization of Paramyxine with Eptatretus. This created homonymy of Paramyxine fernholmi with Eptatretus fernholmi and Paramyxine wisneri with Eptatretus wisneri. In order to resolve this nomenclatural dilemma, we made a more extensive phylogenetic assessment of the Myxinidae and examined the nomenclature of the family. We used 75 sequences (37 of which new for this study) of a 561 bp fragment of the 16S rRNA gene, representing 33 species, and 72 sequences (37 of which new for this study) of a 687 bp fragment of the cytochrome c oxidase subunit I (COI) gene, representing 23 species, to reconstruct the phylogeny of Myxinidae. The monophyly of the subfamily Myxininae, traditionally characterized by having a single pair of external gill openings, was rejected (0.50 Bayesian posterior probability) by the 16S analysis, but supported by the COI and combined COI+16S analyses (0.99 and 0.81 Bpp, respectively). The monophyly of the subfamily Eptatretinae, characterized by having several pairs of external gill openings, was not supported by the 16S analysis and rejected by the COI and combined COI+16S analysis due to the placement of Eptatretus lopheliae as the earliest branch of Myxinidae (0.71 and 0.57 Bpp, respectively). Eptatretus lopheliae and Eptatretus rubicundus formed a monophyletic group and were allocated to a new genus, Rubicundus, characterized by the presence of an elongated tubular nostril and reddish coloration. A new monotypic subfamily, Rubicundinae, was proposed for Rubicundus. The synonymy of the genera Paramyxine and Quadratus with Eptatretus was confirmed. E. fernholmi is renamed Eptatretus luzonicus. Eptatretus wisneri was renamed Eptatretus bobwisneri. Petromyzon cirrhatus Forster, 1801, Homea banksii Fleming, 1822, and Bdellostoma forsteri Müller, 1836 are synonyms, but no type specimens are known to exist. Petromyzon cirrhatus was designated as type species of Eptatretus, conserving present usage. Gastrobranchus dombeyi Shaw, 1804 has priority over other names for Chilean myxinids. Bdellostoma stoutii was designated as type species of Polistotrema Gill. The validity of the Western Atlantic Myxine limosa as distinct from the Eastern Atlantic Myxine glutinosa was confirmed.     

Sodium/calcium selectivity of cloned calcium T-type channels

We studied the peculiarities of permeability with respect to the main extracellular cations, Na⁺ and Ca²⁺, of cloned low-threshold calcium channels (LTCCs) of three subtypes, Ca_v3.1 (α1G), Ca_v3.2 (α 1H), and Ca_v3.3 (α1I), functionally expressed in Xenopus oocytes. In a calcium-free solution containing 100 mM Na⁺ and 5 mM calcium-chelating EGTA buffer (to eliminate residual concentrations of Ca²⁺) we observed considerable integral currents possessing the kinetics of inactivation typical of LTCCs and characterized by reversion potentials of −10 ± 1, −12 ± 1, and −18 ± 2 mV, respectively, for Ca_v3.1, Ca_v3.2, and Ca_v3.3 channels. The presence of Ca²⁺ in the extracellular solution exerted an ambiguous effect on the examined currents. On the one hand, Ca²⁺ effectively blocked the current of monovalent cations through cloned LTCCs (K _d = 2, 10, and 18 μM for currents through channels Ca_v3.1, Ca_v3.2, and Ca_v3.3, respectively). On the other hand, at the concentration of 1 to 100 mM, Ca²⁺ itself functioned as a carrier of the inward current. Despite the fact that the calcium current reached the level of saturation in the presence of 5 mM Ca²⁺ in the external solution, extracellular Na⁺ influenced the permeability of these channels even in the presence of 10 mM Ca²⁺. The Ca_v3.3 channels were more permeable with respect to Na⁺ (P _Ca/P _Na ∼ 21) than Ca_v3.1 and Ca_v3.2 (P _Ca/P _Na ∼ 66). As a whole, our data indicate that cloned LTCCs form multi-ion Ca²⁺-selective pores, as these ions possess a high affinity for certain binding sites. Monovalent cations present together with Ca²⁺ in the external solution modulate the calcium permeability of these channels. Among the above-mentioned subtypes, Ca_v3.3 channels show the minimum selectivity with respect to Ca²⁺ and are most permeable for monovalent cations. Neirofiziologiya/Neurophysiology, Vol. 38, No. 3, pp. 183–192, May–June, 2006.