On the effect of glucono-δ-lactone on the yeast Pichia polymorpha

Investigation has been made into the action of glucono--lactone on living cells of Pichia polymorpha in relation to the uptake of D-(U-¹⁴C) glucose, and the incorporation of (2-¹⁴C) uracil and L-(U-¹⁴C)-threonine into RNA and protein respectively. Other factors such as the action of glucono--lactone on cell morphology and on enzymic synthesis have also been studied. The action of this compound on -glucanase has been found to take place in the hydrolytic power and not in the synthesis.     

The survey of ectoparasites on two species of Triakids (Mustelus mustelus and M. punctulatus) sharks from Tunisian coasts

Between 2018 and 2020, 696 fish belonging to two species of sharks from the Family Triakidae (Mustelus mustelus and M. punctulatus) were collected from the coasts of Tunisia and inspected for parasites. Six copepod taxa (Perissopus dentatus Steenstrup & Lütken, 1861, Eudactylinella alba Wilson, 1932, Kroyeria lineata Van Beneden, 1853, Nesippus orientalis Heller, 1865 and Lernaeopoda galei Krøyer, 1837, Kroyeria sp.), four isopod species (Anilocra physodes (Linnaeus, 1758), Emetha audouini (H. Milne Edwards, 1840), Ceratothoa parallela (Otto, 1828) and Ceratothoa oestroides (Risso, 1816)) and two monogenean species (Erpocotyle sp1. And Erpocotyle sp2) were collected. A large number of global host records was reported, including the occurrence of E. audouini on M. mustelus and of Erpocotyle sp.2 on M. punctulatus. The study of the diversity of parasites per host species revealed that M. mustelus had a higher parasitic richness compared to M. punctulatus. In this study, it was provided for the first records on ectoparasites on Triakidae sharks from Tunisian coasts and their infection indices.     

A high-throughput turbidometric assay for screening inhibitors of Leishmania major protein disulfide isomerase

The use of a high-throughput technique to perform a pilot screen for Leishmania major protein disulfide isomerase (LmPDI) inhibitors identification is reported. In eukaryotic cells, protein disulfide isomerase (PDI) plays a crucial role in protein folding by catalyzing the rearrangement of disulfide bonds in substrate proteins following their synthesis. LmPDI displays similar domain structure organization and functional properties to other PDI family members and is involved in Leishmania virulence. The authors used a method based on the enzyme-catalyzed reduction of insulin in the presence of dithiothreitol. The screen of a small library of 1920 compounds was performed in a 384-well format and led to the identification of 27 compounds with inhibitory activity against LmPDI. The authors further tested the cytotoxicity of these compounds using Jurkat cells as well as their effect on Leishmania donovani amastigotes using high-content analysis. Results show hexachlorophene and a mixture of theaflavin monogallates inhibit Leishmania multiplication in infected macrophages derived from THP-1 cells, although the inhibitory effect on LmPDI enzymatic activity does not necessarily correlate with the antileishmanial activity.     

Mise en évidence d’une flore oxfordienne dans le Sud-Est de la Tunisie : intérêts stratigraphique et paléoécologique

The Late Jurassic - Early Cretaceous series encountered in South-East Tunisia, formely considered globally as Purbecko-Wealdian in age, has the most complete palaeobotanical record of all North-Gondwana. Thanks to the stratigraphical results of the last years and to new field researches, it was possible, for the first time, to evidence an Oxfordian flora at the base of this series (Bir Miteur Formation). This flora, constituted of impressions and axes of Pteridophytes and Gymnosperms, is encountered within different types of plant-bearing levels. The study of deposit facies and sequential analysis of this Oxfordian formation allowed the identification of corresponding sedimentary environments and to infer their palaeoecological meaning. The discovery of exceptional specimens provided new informations about the general morpho-anatomy of Alstaettia, a very peculiar tree fern of a now extinct type. This Oxfordian flora is an interesting key-point in the evolution of North-Gondwanian flora at the Jurassic/Cretaceous transition.     

Use of an inhibitor to identify members of the hormone-sensitive lipase family

Hormone-sensitive lipase (HSL) contributes importantly to the mobilization of fatty acids from the triacylglycerols stored in adipocytes, which provide the main source of energy in mammals. On the basis of amino acid sequence alignments and three-dimensional structures, this enzyme was previously found to be a suitable template for defining a family of serine carboxylester hydrolases. In this study, the HSL family members are characterized rather on the basis of their inhibition by 5-methoxy-3-(4-phenoxyphenyl)-3H-[1,3,4]oxadiazol-2-one (compound 7600). This compound inhibits mammalian HSL as well as other HSL family members, such as EST2 from the thermophilic eubacterium Alicyclobacillus acidocaldarius and AFEST from the hyperthermophilic archaeon Archaeoglobus fulgidus. Various carboxylester hydrolases that are not members of the HSL family were found not to be inhibited by compound 7600 under the same experimental conditions. These include nonlipolytic hydrolases such as Torpedo californica acetylcholinesterase and pig liver esterase, as well as lipolytic hydrolases such as human pancreatic lipase, dog gastric lipase, Thermomyces lanuginosus lipase, and Bacillus subtilis LipA. When vinyl esters were used as substrates, the residual activity of HSL, AFEST, and EST2 decreased with an increase in compound 7600 concentration in the incubation mixture. The inhibitor concentration at which the enzyme activity decreased to 50% after incubation for 5 min was 70, 20, and 15 nM with HSL, AFEST, and EST2, respectively. Treating EST2 and AFEST with the inhibitor resulted in an increase in the molecular mass, as established by performing matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis. This increase in the molecular mass, which corresponds approximately to the molecular mass of the inhibitor, indicates that a covalent enzyme-inhibitor complex has been formed. Surface-enhanced laser desorption ionization time-of-flight mass spectrometry analysis of a trypsin digest of AFEST treated with the inhibitor or not treated showed the occurrence of an increase in the molecular masses of the "GESAGG"-containing peptide, which is compatible with the formation of a covalent complex with the inhibitor.     

赤潮棕囊藻(Phaeocystis globosa)rDNA ITS区序列变异与二级结构分析

测定了南海球形棕囊藻香港株P1、P2和湛江株ZhJ1的rDNAITS区序列(含5.8srDNA),结合Gen Bank的13条同源序列,比对长度为904bp,变异位点271个,简约信息位点221个,平均(A+T)(34.5%)<(G+C)(65.4%).藻株P1、P2和ZhJ1序列存在变异位点20个,序列间相似性为97.9%～98.5%.ITS序列在种间和种内的解析度高于18srDNA和28srDNA基因;构建的NJ树、MP树、贝叶斯推断系统树的结构是一致的,不同种类的棕囊藻单独聚类,不同地理来源的球形棕囊藻混杂分布但相同地理来源的藻株多聚类在一起.RNA二级结构显示,不同藻种间5.8srDNA区结构基本一致,表现出属的特异性;ITS1、2区结构表现较大的种间差异,表明ITS区RNA二级结构可为棕囊藻分类鉴定提供有用的分子结构信息.     

Genetic Diversity and Population Structure of Acacia senegal (L) Willd. in Kenya

The level of genetic diversity and population structure of Acacia senegal variety kerensis in Kenya was examined using seven polymorphic nuclear microsatellite loci and two chloroplast microsatellite loci. In both chloroplast and nuclear datasets, high levels of genetic diversity were found within all populations and genetic differentiation among populations was low, indicating extensive gene flow. Analysis of population structure provided support for the presence of two groups of populations, although all individuals had mixed ancestry. Groups reflected the influence of geography on gene flow, with one representing Rift Valley populations whilst the other represented populations from Eastern Kenya. The similarities between estimates derived from nuclear and chloroplast data suggest highly effective gene dispersal by both pollen and seed in this species, although population structure appears to have been influenced by distributional changes in the past. The few contrasts between the spatial patterns for nuclear and chloroplast data provided additional support for the idea that, having fragmented in the past, groups are now thoroughly mixed as a result of extensive gene flow. For the purposes of conservation and in situ management of genetic resources, sampling could target a few, large populations ideally distributed among the spatial groups identified. This should ensure the majority of extant variation is preserved, and facilitate the investigation of variation in important phenotypic traits and development of breeding populations.     

Dactylogyrids (Monogenoidea: Polyonchoinea) parasitising the gills of snappers (Perciformes: Lutjanidae): Species of Euryhaliotrema Kritsky & Boeger, 2002 from the golden snapper Lutjanus johnii (Bloch) off northern Australia, with a redescription of Euryhaliotrema johni (Tripathi, 1959) and descriptions of two new species

Three species of Euryhaliotrema Kritsky & Boeger, 2002 (Monogenoidea: Dactylogyridae) were collected from the gills of four golden snapper Lutjanus johnii (Bloch) (Lutjanidae) from the marine and brackish waters off Darwin, Northern Territory, Australia. Type-specimens of Ancyrocephalus johni Tripathi, 1959 apparently have not survived and the possibility existed that the species was based on specimens representing more than one species. Euryhaliotrema johni (Tripathi, 1959) (sensu Young, 1968) was redescribed and determined to most likely represent A. johni, originally described from the River Hooghly, Diamond Harbour, India. Two new species were described. Euryhaliotrema longibaculoides n. sp. was most similar to Euryhaliotrema longibaculum (Zhukov, 1976) Kritsky & Boeger, 2002 from Lutjanus spp. from the western Atlantic Ocean. It differed from E. longibaculum by having a male copulatory organ (MCO) with an elongate comparatively delicate shaft and a bulbous base (MCO U- or J-shaped with funnel-shaped base in E. longibaculum). Based on the comparative morphology of the haptoral sclerites, Euryhaliotrema lisae n. sp. was most similar to Euryhaliotrema cryptophallus Kritsky & Yang, 2012 from the gills of the mangrove red snapper Lutjanus argentimaculatus (Forsskål) from the South China Sea. Euryhaliotrema lisae differed from E. cryptophallus by having a copulatory complex with an obvious weakly sclerotised J-shaped MCO (MCO cryptic, delicate, and with a shaft comprising about one counterclockwise ring in E. cryptophallus).     

Species traits and climate velocity explain geographic range shifts in an ocean‐warming hotspot

Species' ranges are shifting globally in response to climate warming, with substantial variability among taxa, even within regions. Relationships between range dynamics and intrinsic species traits may be particularly apparent in the ocean, where temperature more directly shapes species' distributions. Here, we test for a role of species traits and climate velocity in driving range extensions in the ocean‐warming hotspot of southeast Australia. Climate velocity explained some variation in range shifts, however, including species traits more than doubled the variation explained. Swimming ability, omnivory and latitudinal range size all had positive relationships with range extension rate, supporting hypotheses that increased dispersal capacity and ecological generalism promote extensions. We find independent support for the hypothesis that species with narrow latitudinal ranges are limited by factors other than climate. Our findings suggest that small‐ranging species are in double jeopardy, with limited ability to escape warming and greater intrinsic vulnerability to stochastic disturbances.     

Proteome analysis reveals novel proteins associated with proliferation and differentiation of the colorectal cancer cell line Caco-2

Here, we describe a proteomics approach to study protein expression changes in differentiating Caco-2 cells. Caco-2 is a colorectal carcinoma cell line, which upon differentiation loses its tumorigenic phenotype and displays characteristics of mature enterocytes, including brush borders with microvilli. Cells were grown in culture flasks and harvested at different stages of differentiation (days post-confluence: -3, 0, 3, 7, 10, 14, and 18). Two-dimensional gel electrophoresis was used to analyse proteome changes. Approximately 1400 protein spots were detected within the Caco-2 proteome, within the pH 4-7 range. Two-dimensional gel electrophoresis allowed for the detection of 18 proteins from which the levels of expression were found to be associated with differentiation. Of these proteins, 11 were identified by means of MALDI-TOF or NANO-ESI-MS/MS mass spectrometry and include liver fatty acid binding protein (FABL), three forms of alpha-enolase (ENOA), nucleoside diphosphate kinase A (NDKA), cofilin-1 (COF1), translationally controlled tumour protein (TCTP), mitochondrial 60-kDa heat shock protein (CH60), probable protein disulfide isomerase (ER60), creatine kinase B (KCRB), and glutathione S-transferase alpha (GTA1). Thus, proteomics revealed that the differentiation-related change in phenotype of Caco-2 involves changes in a variety of distinct biochemical pathways. Some of these proteins have not been shown before to be associated with Caco-2 differentiation (ER60; COF1; CH60; NDKA; TCTP and ENOA). Therefore, processes related to protein folding and disulfide bridge formation, cytoskeleton formation and maintenance, nucleotide metabolism, glycolysis as well as tumorigenesis-associated proteins may be involved in Caco-2 differentiation. Changes in the expression of CH60, TCTP, GTA1, NDKA, and FABL have also been reported to be associated with in vivo colon carcinogenesis. These findings illustrate that a combination of proteomics and cell culture is a useful approach to find markers for Caco-2 differentiation, which could contribute to the comprehension of the process of colon carcinogenesis.