Metabolic activation of selected aromatic amines and polycyclic hydrocarbons by isolated subcellular fractions of Drosophila melanogaster

The applicability of microsomal preparations from Drosophilamelanogaster as the metabolic factor in the Salmonella mutagenicity assay with strains TA98 and TA100 was evaluated. Isolated cellular fractions (S27) from PB-pretreated flies activated N-acetyl-2-aminofluorene (2-AAF), N-hydroxy-N-aceyl-2-aminofluorene (N-OH-AAF), benzo[a]pyrene (BP), 9,10-dimethylanthracene (DA) and 2 -naphythylamine (NA)_into mutagenic metabolites, 7,-12-Dimethylbenz[a]-anthracene (DMBA) was ineffective under the conditions of the test.This study was performed in an effort to determine optimal conditions for activating, by Drosophila enzymes, aromatic amines and polycyclic hydrocarbons, with 2-AAF and BP as model mutagens. The following alterations improved the sensitivity of this combined Salmonella/Drosophila assay. (1) Incubation of the plates at 25°C for 1 night instead of permanent exposure at 37°C. (2) Isolation of S27 fractions instead of the conventional S9, because 9000 × g was not sufficient tio spin down Drosphila mitochondria.     

Full characterization of PDX, a neuroprotectin/protectin D1 isomer, which inhibits blood platelet aggregation

Our study aimed to establish the complete structure of the main dihydroxy conjugated triene issued from the lipoxygenation (soybean enzyme) of docosahexaenoic acid, named PDX, an isomer of protectin/neuroprotectin D1 (PD1/NPD1) described by Bazan and Serhan. NMR approaches and other chemical characterization (e.g. GC-MS, HPLC and LC-MS/MS) indicated that PDX is 10(S),17(S)-dihydroxy-docosahexa-4Z,7Z,11E,13Z,15E,19Z-enoic acid. The use of ¹⁸O₂ and mass spectrometry showed that PDX is a double lipoxygenation product. Its structure differs from PD1, with E,Z,E geometry (PDX) instead of E,E,Z (PD1) and S configuration at carbon 10 instead of R. PDX inhibits human blood platelet aggregation at sub-micromolar concentrations.     

Halichoeres bleekeri (steindachner & döderlein), a valid japanese species of labrid fish, distinct fromH. tenuispinis (günther) from china

Halichoeres bleekeri (Steindachner &; Döderlein, 1887), previously regarded as a junior synonym ofH. tenuispinis (Günther), is a valid species. It is distinct fromH. tenuispinis in nearly always having 13 instead of 14 pectoral rays. 11–15 instead of 7–11 suborbital pores, longer dorsal soft rays (1.95–2.3 in head length vs 2.25–2.4 fortenuispinis). and some features of color such as the absence of a dark spot at the upper base of the caudal fin of the female.H. tremebundus Jordan and Snyder, 1902 is a synonym ofH. bleekeri based on the female form, andArtisia festiva de Beaufort is a synonym ofH. tenuispinis, also described from the female phase,H. bleekeri is known from Korea and in Japan from Tokyo to the Izu Islands.H. tenuispinis is recorded from Hong Kong and Xiamen. China, and Taiwan. Records ofH. tenuispinis from the Philippines by Fowler and Bean (1928) represent misidentifications ofH. papilionaceus (Valenciennes).     

Differential detection of phosphorylated glycogen synthase kinase 3 alpha and beta depending on blocking conditions

It is generally recommended that immunoblots using phosphospecific antibodies be performed using bovine serum albumin (BSA) instead of milk. There are two subtypes of glycogen synthase kinase 3 (GSK3), GSK3 α and GSK3 β, with apparent molecular weights of 51 and 49 kDa, respectively. Here we show that immunoblots performed using 5% milk allow the detection of both phosphorylated GSK3 α and phosphorylated GSK3 β, whereas only phosphorylated GSK3 α is visible when immunoblots are performed using 3% BSA.     

A redescription of the presumed associated Caligopsyllus primus Kunz, 1975 (Harpacticoida,Paramesochridae) with emphasis on its phylogenetic affinity with Apodopsyllus Kunz, 1962

Caligopsyllus primus Kunz, 1975, reported from tidal pools near East London (South Africa), is redescribed and figured on the basis of type material. An amended diagnosis of the genus Caligopsyllus Kunz, 1975 is presented. Cladistic analysis falsified the hypothesis of a close relationship between Caligopsyllus and Kliopsyllus Kunz, 1962, favouring instead a link with Apodopsyllus Kunz, 1962. The morphological modifications are discussed within the context of a possible ecto-associated life-style. Finally, a standard format of abbreviations and terminology is proposed for the paramesochrid caudal ramus.     

Zur primärstruktur des cytochromes c des steppenzebras (Equus quagga Boehmi)

Cytochrome c of the zebra (Equus guagga Boehmi) differs from horse cytochrome c in having in its polypeptide chain one serine residue instead of none and 9 threonine residues instead of 10. This replacement has been localised at position 47 of the sequence.     

Cyphastrea kausti sp. n. (Cnidaria,Anthozoa, Scleractinia), a new species of reef coral from the Red Sea

A new scleractinian coral species, Cyphastrea kausti sp. n., is described from 13 specimens from the Red Sea. It is characterised by the presence of eight primary septa, unlike the other species of the genus, which have six, ten or 12 primary septa. The new species has morphological affinities with Cyphastrea microphthalma, from which it can be distinguished by the lower number of septa (on average eight instead of ten), and smaller calices and corallites. This species was observed in the northern and central Red Sea and appears to be absent from the southern Red Sea.     

Some notes on Marattia anglica from the Jurassic of Yorkshire

In this paper the spores of the Jurassic fern Marattia anglica are redescribed. It appeared that at least 30% of the spores have a more or less trilete, instead of the usual monolete, laesura. In Marattia muensteri, M. hoerensis and M. crenulata (three species from the Jurassic), trilete spores also appeared to occur, although in much lower percentages than in M. anglica. The spores of M. anglica are compared with recent Marattia spores and with Jurassic dispersed spores.Furthermore, the systematic position of M. anglica and other Jurassic Marattia species is discussed.     

A new species of <Emphasis Type="Italic">Tinrovia</Emphasis> Mamaev, 1987 (Monogenea: Microcotylidae) from the deep-sea fish <Emphasis Type="Italic">Notacanthus bonaparte</Emphasis> Risso (Notacanthiformes: Notacanthidae) in the Western Mediterranean and the North East Atlantic

A new microcotylid, Tinrovia mamaevi n. sp. (Monogenea: Polyopisthocotylea), is described from the gills of Notacanthus bonaparte Risso (Notacanthiformes: Notacanthidae), sampled in the Western Mediterranean and North East Atlantic. This species is allocated to the subfamily Syncoelicotylinae Mamaev & Zubchenko, 1978 due to the possession of a symmetrical haptor with two separate frills. The clamps in T. mamaevi n. sp. are of the “microcotylid” type, arranged in two distinct lateral haptoral frills; the genital atrium and the copulatory organ are armed and the vaginal pore is unarmed. The new species differs from the type- and only species of the genus, T. papiliocauda Mamaev, 1987, in having a shorter and narrower haptor with a smaller number of clamps. Clamps are also smaller in the new species, testes are more numerous, the genital atrium is smaller, divided into two lateral lobes (instead of five) with a smaller number of spines and the eggs have a short and a long filament (instead of two short filaments). The host species and locality of T. mamaevi n. sp. also differ as T. papiliocauda which was recorded in Notacanthus sexspinis Richardson from the South Pacific.     

Characterization and anti-Aspergillus flavus impact of nanoparticles synthesized by Penicillium citrinum

This work was conducted to evaluate the ability of grape molding fungus; Penicillium citrinum to synthesize silver nanoparticles (Ag NPs). The potency of biosynthesized Ag NPs was checked against the aflatoxigenic Aspergillus flavus var. columnaris, isolated from sorghum grains. Biosynthesized Ag NPs were characterized and confirmed in different ways. X ray diffraction (XRD), Energy Dispersive Spectroscopy (EDS), Transmission Electron Microscopy (TEM) and optical absorption measurements confirmed the bio-synthesis of Ag NPs. The in vitro antifungal investigation showed that biosynthesized Ag NPs were capable of inhibiting the growth of aflatoxigenic A. flavus var. columnaris. Utilization of plant pathogenic fungi in the Ag NPs biosynthesis as well as the use of bio-Ag NPs to control fungal plant diseases instead of chemicals is promising. Further work is needed to confirm the efficacy of the bio-Ag NPs against different mycotoxigenic fungi and to determine the potent applicable doses.     

Promoter hypermethylation using 24-gene array in early head and neck cancer: Better outcome in oral than in oropharyngeal cancer

Silencing of tumor suppressor genes (TSGs) by DNA promoter hypermethylation is an early event in carcinogenesis and a potential target for personalized cancer treatment. In head and neck cancer, little is known about the role of promoter hypermethylation in survival. Using methylation specific multiplex ligation-dependent probe amplification (MS-MLPA) we investigated the role of promoter hypermethylation of 24 well-described genes (some of which are classic TSGs), which are frequently methylated in different cancer types, in 166 HPV-negative early oral squamous cell carcinomas (OSCC), and 51 HPV-negative early oropharyngeal squamous cell carcinomas (OPSCC) in relation to clinicopathological features and survival. Early OSCC showed frequent promoter hypermethylation in RARB (31% of cases), CHFR (20%), CDH13 (13%), DAPK1 (12%), and APC (10%). More hypermethylation (≥ 2 genes) independently correlated with improved disease specific survival (hazard ratio 0.17, P = 0.014) in early OSCC and could therefore be used as prognostic biomarker. Early OPSCCs showed more hypermethylation of CDH13 (58%), TP73 (14%), and total hypermethylated genes. Hypermethylation of two or more genes has a significantly different effect on survival in OPSCC compared with OSCC, with a trend toward worse instead of better survival. This could have a biological explanation, which deserves further investigation and could possibly lead to more stratified treatment in the future.     

Das nördlichste Vorkommen paläogener Säugetiere in Europa

The localities Dalum and Osteroden near Fürstenau (Lower Saxony, NW-Germany) represent the northern-most occurrences of Palaeogene mammals in Europe.Eurohippus parvulus messelensis at Dalum indicates mammal zone MP 11, hence lower Geiseltalian (middle Eocene), instead of MP 10, ?Cuisian“ (= Grauvian, early Eocene) asTobien (1986) assumed. The same holds true for an upper molar from Balegem (Belgium), whileArctocyon primaevus as well asA. matthesi from Dalum andA. matthesi from Osteroden are evidently reworked from the late Paleocene.     

Biological activity of endomorphin and [Dmt1]endomorphin analogs with six-membered proline surrogates in position 2

Endogenous μ-opioid receptor (MOR) selective peptides, endomorphin-1 (EM-1) and endomorphin-2 (EM-2), unlike so called ‘typical opioids’, are characterized by the presence of Pro² residue, which is a spacer connecting aromatic pharmacophoric residues. In order to investigate structural requirements for position 2, we synthesized endomorphin analogs incorporating, instead of Pro, unnatural amino acids with six-membered heterocyclic rings, such as piperidine 2-, 3- or 4-carboxylic acids (Pip, Nip and Inp, respectively). (R)-Nip residue turned out to be favourable for improving MOR affinity. Introduction of 2′,6′-dimethyltyrosine (Dmt) instead of Tyr¹ led to obtaining [Dmt¹, (R)-Nip²]EM-2 which showed exceptional MOR affinity and high stability against enzymatic degradation in rat brain homogenate. In in vivo hot-plate test in mice, this analog given intracerebroventicularly (i.c.v.), produced profound supraspinal analgesia, being much more potent than EM-2. The antinociceptive effect of this analog lasted about 170 min and was almost completely reversed by β-funaltrexamine (β-FNA), a selective MOR antagonist.     

DNA polymerases β and λ do not directly affect Ig variable region somatic hypermutation although their absence reduces the frequency of mutations

During somatic hypermutation (SHM) of antibody variable (V) region genes, activation-induced cytidine deaminase (AID) converts dC to dU, and dUs can either be excised by uracil DNA glycosylase (UNG), by mismatch repair, or replicated over. If UNG excises the dU, the abasic site could be cleaved by AP-endonuclease (APE), introducing the single-strand DNA breaks (SSBs) required for generating mutations at A:T bp, which are known to depend upon mismatch repair and DNA Pol η. DNA Pol β or λ could instead repair the lesion correctly. To assess the involvement of Pols β and λ in SHM of antibody genes, we analyzed mutations in the VDJh4 3′ flanking region in Peyer's patch germinal center (GC) B cells from polβ^?/?polλ^?/?, polλ^?/?, and polβ^?/? mice. We find that deficiency of either or both polymerases results in a modest but significant decrease in V region SHM, with Pol β having a greater effect, but there is no effect on mutation specificity, suggesting they have no direct role in SHM. Instead, the effect on SHM appears to be due to a role for these enzymes in GC B cell proliferation or viability. The results suggest that the BER pathway is not important during V region SHM for generating mutations at A:T bp. Furthermore, this implies that most of the SSBs required for Pol η to enter and create A:T mutations are likely generated during replication instead. These results contrast with the inhibitory effect of Pol β on mutations at the Ig Sμ locus, Sμ DSBs and class switch recombination (CSR) reported previously. We show here that B cells deficient in Pol λ or both Pol β and λ proliferate normally in culture and undergo slightly elevated CSR, as shown previously for Pol β-deficient B cells.     

Caging experiment: Relationship between mesh size and artifacts

In marine sediments, cages are widely used to detect effects of predation on infauna. Artifacts due to the use of cages have been, instead, poorly estimated. In this study we have analysed the changes in macrofauna and in sediment composition when cages of different mesh sizes are used to exclude predators. In a shallow micro-tidal flat, the effect of three mesh sizes (2, 6 and 18 mm) was tested on sediment and macrofauna, using cages that excluded predators (EC), partial cages (PC) and natural sediment (un-caged plots, UC). PC allowed the predators to feed on the sediment, but modified the sediment similarly to the complete cages. Changes were measured for the abundance and species composition of macrofauna and for sediment total organic matter (TOM), chlorophyll-a, proteins and water content.The use of cages did not alter TOM and water content, whereas chlorophyll-a and phaeopigments were more abundant under the cages. The deep-burrowing shrimp Upogebia pusilla also increased under cages of any mesh-size. There was, instead, an effect on the abundance of macrofauna only when the smallest mesh size was used. Surprisingly, the exclusion of predators decreased the abundances of Tanais dulongii, Oligochaetes, Brania oculata, whereas the abundance of Aphelochaeta marionii changed when both PC and EC were put in the sediment. Collectively, all these findings indicated that for surface animals the use of small mesh sizes can bias the interpretation of results.     

Detection of p16 promotor hypermethylation in “Maras powder” and tobacco users

Background: A plant powder called “Maras powder” is widely used instead of cigarette smoking in the South-Eastern region of Turkey. It has been confirmed that this powder comprises tobacco Nicotiana rustica L. Methods: The aim of this study was to assess the effect of Maras powder and cigarette smoking on the P16 promotor hypermethylation. Twenty-two Maras powder users (Group I), 12 cigarette smokers (Group II), and 16 healthy controls who neither smoked nor used Maras powder (Group III) were included in the study. Hypermethylation of the P16 gene was examined using methylation-specific PCR (MSP) method in the blood of the three groups. Results: Aberrant P16 methylation was found in 7 of the 22 (31.8%) in Group I, in 3 of 12 (25%) in Group II, and in 1 of 16 (6.25%) in Group III. Conclusion: Maras powder may be as harmful as cigarette smoking, leading to hypermethylation in P16 and warrants detailed studies on this subject.     

Antigenic polysaccharides of bacteria: 42. Structures of O-polysaccharides from two Shigella dysenteriae type 8 strains

The structure of the O-polysaccharide (O-antigen) from Shigella dysenteriae type 8 bacteria (strain 599) was corrected using modern NMR techniques (structure 1). The revisions concerned the position of the Glc residue (in the main, but not the side chain), the site of its substitution, and the configuration of the O-glycoside linkage of the GlcNAc residue. The S. dysenteriae type 8 bacterium (strain G1221), the second investigated representative, was found to produce another structural variant of the O-polysaccharide. It contains GlcNAc instead of the Glc residue in the main chain (structure 2). This data may lead to approval of division of S. dysenteriae type 8 into two subtypes:      

Conformational studies on complexes of a diimine containing a (CH2)2 spacer: crystal structures of a double-stranded Zn(II) meso-helicate and an enantiopure Δ-Cu(II) monohelicate

Ni(II), Cu(II), Zn(II) and Cd(II) complexes of an N₄-donor Schiff base, containing (CH₂)₂ as spacer, have been prepared. The X-ray crystal structures of monohelical Ni(ETs) · H₂O and the homochirally crystallised Δ-Cu(ETs), as well as the meso-helicate Zn₂(ETs)₂ · MeCN [H₂ETs: N,N^′-bis(2-tosylaminobenzylidene)-1,2-diaminoethane] have been solved. In the latter, the ligand behaves as bis-bidentate, displaying a “C”-type arrangement, instead of the typical “S”-type fashion present in bis-helical dinuclear complexes.     

Protection against Helicobacter pylori infection by a trivalent fusion vaccine based on a fragment of urease B-UreB414

A multivalent fusion vaccine is a promising option for protection against Helicobacter pylori infection. In this study, UreB414 was identified as an antigenic fragment of urease B subunit (UreB) and it induced an antibody inhibiting urease activity. Immunization with UreB414 partially protected mice from H. pylori infection. Furthermore, a trivalent fusion vaccine was constructed by genetically linking heat shock protein A (HspA), H. pylori adhesin A (HpaA), and UreB414, resulting in recombinant HspA-HpaA-UreB414 (rHHU). Its protective effect against H. pylori infection was tested in BALB/c mice. Oral administration of rHHU significantly protected mice from H. pylori infection, which was associated with H. pylori-specific antibody production and Th1/Th2-type immune responses. The results show that a trivalent fusion vaccine efficiently combats H. pylori infection, and that an antigenic fragment of the protein can be used instead of the whole protein to construct a multivalent vaccine.