Induction of Phage Production in the Lysogenic Escherichia coli by Hydroxyurea

1) Hydroxyurea, a reversible DNA synthesis inhibitor, was used to study the mechanism of prophage λ induction in Escherichia coli K12. Induction of prophage was judged on two criteria: increase of phage-producing cells and loss of colony-forming ability of the cells. 2) Hydroxyurea induced an increase of phage-producing cells only in lysogenic strains known to be inducible with ultraviolet irradiation for prophage development and not in strains such as E. coli K12 (λind^–) or E. coli K12 recA (λ⁺). 3) When protein synthesis was inhibited, hydroxyurea did not increase phage-producing cells of lysogenic strains; it showed a bacteriocidal effect on lysogenic recA⁺ strains, but not on nonlysogenic strains. 4) The sensitivity of E. coli K12 recA to hydroxyurea was independent of whether or not the cells were lysogenic. 5) From the results it is suggested that certain steps leading to loss of colony-forming ability (i.e. prophage induction) do not require de novo protein synthesis but require the presence of the host recA⁺ gene.     

Proposed mechanism of inactivating Escherichia coli O157:H7 by ultra‐high pressure in combination with tert‐butylhydroquinone

Aims: Investigating mechanisms of lethality enhancement when Escherichia coli O157:H7, and selected E. coli mutants, were exposed to tert‐butylhydroquinone (TBHQ) during ultra‐high pressure (UHP) treatment. Methods and Results: Escherichia coli O157:H7 EDL‐933, and 14 E. coli K12 strains with mutations in selected genes, were treated with dimethyl sulfoxide solution of TBHQ (15–30 ppm), and processed with UHP (400 MPa, 23 ± 2°C for 5 min). Treatment of wild‐type E. coli strains with UHP alone inactivated 2·4–3·7 log CFU ml^?1, whereas presence of TBHQ increased UHP lethality by 1·1–6·2 log CFU ml^?1; TBHQ without pressure was minimally lethal (0–0·6 log reduction). Response of E. coli K12 mutants to these treatments suggests that iron–sulfur cluster‐containing proteins ([Fe–S]‐proteins), particularly those related to the sulfur mobilization (SUF system), nitrate metabolism, and intracellular redox potential, are critical to the UHP–TBHQ synergy against E. coli. Mutations in genes maintaining redox homeostasis and anaerobic metabolism were associated with UHP–TBHQ resistance. Conclusions: The redox cycling activity of cellular [Fe–S]‐proteins may oxidize TBHQ, potentially leading to the generation of bactericidal reactive oxygen species. Significance and Impact of the Study: A mechanism is proposed for the enhanced lethality of UHP by TBHQ against E. coli O157:H7. The results may benefit food processors using UHP–based preservation, and biologists interested in piezophilic micro‐organisms.     

Earliest ontogeny of Early Palaeozoic Craniiformea: compelling evidence for lecithotrophy

Popov, L.E., Bassett, M.G. & Holmer, L.E. 2012: Earliest ontogeny of Early Palaeozoic Craniiformea: compelling evidence for lecithotrophy. Lethaia, Vol. 45, pp. 566–573. The early ontogeny of Palaeozoic Craniiformea (Brachiopoda) remains controversial, with conflicting reports of evidence indicating lecithotrophic versus planktotrophic larval stages. Further compelling evidence for lecithotrophy in Palaeozoic craniiforms is described here. Newly obtained, well‐preserved Silurian specimens of craniiforms, including Craniops (Craniopsida), and Lepidocrania? and Orthisocrania (Craniida) from Gotland and the St. Petersburg region, form the basis for this study. The new material demonstrates that the characters of shell structure and shell formation provide evidence of early differentiation of an adult dorsal mantle, and the presence of a distinctive primary layer with a characteristic lath‐like pattern indicates that these craniiforms underwent a lecithotrophic larval stage, more or less identical to that of living. □Novocrania. Brachiopoda, Craniiformea, ontogeny, phylogeny, Early Palaeozoic.     

Recognition of a new species of Kallymenia (Gigartinales,Rhodophyta) from Croatia (Mediterranean Sea) based on morphology and DNA barcode

The Mediterranean red algal flora is diverse but current knowledge of its diversity is at best fragmentary. Here, a new species of Kallymenia from Croatia is described based on morphological and molecular data. Members of the genus Kallymenia share similar morphology making their generic identification relatively easy, whereas species level identification is notoriously difficult. In this paper an integrative systematics study using three gene markers, cox1 (COI), rbcL and nuclear LSU, allowed us to (i) confirm the identity of four existing members of this genus inhabiting the Mediterranean Sea: K. feldmannii, K. lacerata, K. patens and K. requienii; (ii) detect the presence of K. reniformis only in the Atlantic, and (iii) reveal the presence of a new member of Kallymenia from the Mediterranean Sea, K. ercegovicii Vergés & Le Gall, sp. nov., which is described based on distinctive morphological and molecular characters. Kallymenia ercegovicii is distinguished, using three molecular markers, from all Kallymenia species for which these sequences are available. Morphologically, the new species can be distinguished from other polycarpogonial Kallymenia by a set of characters including a deeply lobed thallus, large inner cortical cells, stellate but non-glanglionic medullary cells and non-ostiolate cystocarps surrounded by a filamentous net composed of elongated cells forming fascicles. An initial phylogeny of the genus Kallymenia was inferred from cox1 (COI), rbcL and nuclear LSU sequences. Kallymenia ercegovicii was resolved with full support within the same lineage as K. reniformis (the generitype), K. feldmannii and K. patens, whereas K. lacerata and K. requienii were only distantly related.     

The cranial morphology of Kayentachelys,an Early Jurassic cryptodire,and the early history of turtles

Gaffney, E.S. and Jenkins, F.A., Jr. 2010. The cranial morphology of Kayentachelys, an Early Jurassic cryptodire, and the early history of turtles. — Acta Zoologica (Stockholm) 91 : 335–368 The skull morphology of Kayentachelys aprix Gaffney et al., 1987 , a turtle from the Early Jurassic Kayenta Fm of northern Arizona, demonstrates the presence of cryptodiran synapomorphies in agreement with Gaffney et al. (1987, 1991, 2007) , and contrary to the conclusions of Sterli and Joyce (2007) , Joyce (2007) , Sterli (2008) , and Anquetin et al. (2008) . Specific characters found in Kayentachelys and diagnostic of cryptodires include the processus trochlearis oticum, the curved processus pterygoideus externus with a vertical plate, and the prefrontal–vomer contact, which are confirmed as absent in the outgroups, specifically the Late Triassic Proganochelys. The Joyce (2007) analysis suffers from the reduction of the signal from skull characters, with a consequently greater reliance on shell characters, resulting in pleurodires being resolved at various positions within the cryptodires. Kayentachelys reveals what a primitive cryptodire would be expected to look like: a combination of primitive and derived characters, with the fewer derived characters providing the best test of its relationships to other turtles. Although incompletely known, the Mid‐Late Jurassic Condorchelys, Heckeremys, and Eileanchelys may be early cryptodires close to Kayentachelys. We confirm the Late Triassic Proterochersis as a pleurodire, dating the pleurodire–cryptodire split as Late Triassic or earlier.     

β-Oxidation-mediated resistance ofEscherichia coli to inhibition by long-chain fatty acids

Decanoate exerted a stronger bactericidal action onEscherichia coli K76 (-oxidation negative) than on its -oxidation constitutive parent,E. coli K113; both strains had a normal lipopolysaccharide layer.E. coli RC59, a mutant carrying a defective lipopolysaccharide layer, was more sensitive than its parent,E. coli O111B4, to the growth inhibition by decanoate. Oleate did not affectE. coli RC59 viability, but washing of the cells rendered them sensitive.E. coli RC59 was unable to degrade decanoate and oleate. A mutant of this strain was isolated (E. coli ER20) which retained the defective lipopolysaccharide but was able to metabolize oleic acid. When the killing effect of oleate was assayed on washed cells ofE. coli O111B4, RC59, and ER20, a positive correlation between -oxidation activity and the number of surviving viable cells was found. The ability of Gram-negative organisms to withstand the toxic effect of fatty acids was attributed to the presence of a lipolysaccharide (LPS) layer in the cell envelope. An additional mechanism of resistance is suggested by the results in this report which show that the ability to oxidize fatty acids contributes to resistance.     

TAXONOMIC REVISIONS OF MORPHOLOGICALLY SIMILAR SPECIES FROM TWO EUGLENOID GENERA: EUGLENA (E. GRANULATA AND E. VELATA) AND EUGLENARIA (EU. ANABAENA,EU. CAUDATA,AND EU. CLAVATA)1

The establishment of epitypes (together with the emended diagnoses) for three species of Euglenaria Karnkowska, E. W. Linton et Kwiatowski [Eu. anabaena (Mainx) Karnkowska et E. W. Linton; Eu. caudata (Hübner) Karnkowska et E. W. Linton; and Eu. clavata (Skuja) Karnkowska et E. W. Linton] and two species of Euglena Ehrenberg [E. granulata (Klebs) Schmitz and E. velata Klebs] was achieved due to literature studies, verification of morphological diagnostic features (cell size, cell shape, number of chloroplasts, the presence of mucocysts), as well as molecular characters (SSU rDNA). Now all these species are easy to identify and distinguish, despite their high morphological similarity, that is, spindle‐shaped (or cylindrically spindle‐shaped) cells and parietal, lobed chloroplasts with a single pyrenoid, accompanied by bilateral paramylon caps located on both sides of the chloroplast. E. granulata is the only species in this group that has spherical mucocysts. E. velata is distinguished by the largest cells (90–150 μm) and has the highest number of chloroplasts (>30). Eu. anabaena has the fewest chloroplasts (usually 3–6), and its cells are always (whether the organism is swimming or not) spindle‐shaped or cylindrically spindle‐shaped, in contrast to the cells of Eu. clavata, which are club‐shaped (clavate) while swimming and only after stopping change to resemble the shape of a spindle or a cylindrical spindle; Eu. clavata has numerous chloroplasts (15–20). Eu. caudata is characterized by asymmetrical spindle‐shaped (fusiform) cells, that is, with an elongated rear section and a shorter front section; the number of chloroplasts normally ranges from 7 to 15.     

Whole cell biosynthetic activity of Komagataella phaffii (Pichia pastoris) GS115 strains engineered with transgenes encoding Chromobacterium violaceum ω-transaminase alone or combined with native transketolase

Whole cell biocatalysis is an ideal tool for biotransformations that demand enzyme regeneration or robustness to fluctuating pH, osmolarity and biocontaminant load in feedstocks. The methylotrophic yeast Komagataella phaffii is an attractive alternative to Escherichia coli for whole cell biocatalysis due to its genetic tractability and capacity to grow to up to 60% wet cell weight by volume. We sought to exploit high cell density K. phaffii to intensify whole-cell chiral amino-alcohol (CAA) biosynthesis. We engineered two novel K. phaffii GS115 strains: one by inserting a Chromobacterium violaceum ω-transaminase CV2025 transgene, for strain PpTAmCV708, and a second strain, PpTAm-TK16, by also inserting the same CV2025 transgene plus a second transgene for a native transketolase. At high cell density, both strains tolerated high substrate concentrations. When fed three low cost substrates, 200 mM glycolaldehyde, 200 mM hydroxypyruvate and 150 mM methylbenzylamine, PpTAm-TK16 whole cells achieved 0.29 g L⁻¹ hr⁻¹ space–time yield of the acetophenone by-product, a 49-fold increase of the highest levels reported for E. coli whole cells harboring the equivalent pathway. When fed only the low-cost substrate, 150 mM methylbenzylamine, strain PpTAmCV708 achieved a 105-fold increase of reported E. coli whole cell biocatalysis performance, with a space–time yield of 0.62 g L⁻¹ hr⁻¹ of the CAA, 2-amino-1,3,4-butanetriol (ABT). The rapid growth and high biomass characteristics of K. phaffii were successfully exploited for production of ABT by whole-cell biocatalysis at higher levels than the previously achieved with E. coli in the presence of the same substrates.     

A taxonomic study of Japanese Scytosiphon (Scytosiphonales, Phaeophyceae), including two new species

Morphological and culture studies were carried out on the genus Scytosiphon (Scytosiphonales, Phaeophyceae) growing in Japan. Morphologies of plurilocular zo-idangia and sporophytic thalli were found to be useful as taxonomic characters. Two types of plurilocular zo-idangial sori were recognized: (i) loosely coherent plurilocular zoidangia without a cuticular layer (loose type); (ii) tightly coherent plurilocular zoidangia with a cuticular layer (coherent type). The sporophytic thallus morphology was different among species in general appearance (tufted or discoid), degree of cohesion of erect filaments, and presence or absence of paraphyses. Two new species are described, Scytosiphon gracilis sp. nov. and Scytosiphon tenellus sp. nov, Scytosiphon gracilis is distinguished from other species by a thin medullary layer, coherent plurilocular zoidangia, no ascocysts associated with plurilocular zoidangia, and Compsonema-like sporophytic thalli. Scytosiphon tenellus is characterized by a thin medullary layer, coherent plurilocular zoidangia, ascocysts among plurilocular zoidangial sori, and Stragularia-like sporophytic thalli. Scytosiphon lo-mentaria (Lyngbye) Link is characterized by a constricted gametophytic thallus, loose plurilocular zoidangia, the presence of ascocysts, and sporophytic thaili identified with Microspongium gelatlnosum Reinke.     

The first dinosaur egg from the Lower Cretaceous of Western Siberia,Russia

The Lower Cretaceous Ilek Formation in Western Siberia (Russia) has yielded various vertebrate fossils, including skeletal remains of dinosaurs. Here we report on a fragmentary theropod egg from the vertebrate locality Shestakovo 3 of the Ilek Formation in Kemerovo Province. We assign the specimen to the oogenus Prismatoolithus (oofamily Prismatoolithidae) as Prismatoolithus ilekensis oosp. nov., on the basis of the following unique combination of characters: ovoid-shaped egg; thin eggshell 300–330 μm thick; angustiprismatic morphotype; eggshell with three different layers; gradual transition between mammillary layer and prismatic layer; abrupt contact between prismatic layer and external layer; mammillary layer to prismatic layer to external layer thickness ratio is 1:3:0.6; prismatic layer with ill-defined squamatic texture; angusticanaliculate pore system; and smooth outer surface. Like other Early Creataceous Prismatoolithus, the egg of Prismatoolithus ilekensis oosp. nov. was laid by a small bodied theropod dinosaur (troodontid or primitive bird) and this taxonomic attribution is supported by results of our phylogenetic analysis. Prismatoolithus ilekensis oosp. nov. is the first Early Cretaceous ootaxon from Russia.

urn:lsid:zoobank.org:act:734EAD40-86C3-488B-A61E-B5FF7378BC0E     

A phi Layer in Roots of Ceratonia siliqua L

The central cylinder of the primary root of the carob tree (Ceratonia siliqua) is encircled by a layer of cells with wall thickenings, known as a phi (φ) cell layer. The development of the φ layer and the chemical composition of the cell wall thickenings have been studied in roots of C. siliqua. The results reveal the presence of condensed tannins in the mature phi thickenings and that the development of the φ layer is asynchronous: at 0–1 cm from the root tip φ thickenings appear before endodermis differentiation at the sites opposite phloem, at 1–4 cm new φ thickenings are developed at the sites opposite xylem, at 4–7 cm the φ layer consists of two layers of cells and it completely encloses the central cylinder.     

The lungs of Polypterus senegalus and Erpetoichthys calabaricus: Insights into the structure and functional distribution of the pulmonary epithelial cells

The present article is a comparative, structural study of the lung of Polypterus senegalus and Erpetoichthys calabaricus , two species representative of the two genera that constitute the Polypteriformes. The lung of the two species is an asymmetric, bi‐lobed organ that arises from a slit‐like opening in the ventral side of the pharynx. The wall is organized into layers, being thicker in P. senegalus . The inner epithelium contains ciliated and non‐ciliated bands. The latter constitute the respiratory surface and are wider in E. calabaricus . The air‐blood barrier is thin and uniform in P. senegalus and thicker and irregular in E. calabaricus . In the two species, the ciliated areas contain ciliated cells, mucous cells and cells with lamellar bodies. Additionally, P. senegalus contains polymorphous granular cells (PGCs) and neuroendocrine cells (NECs) while E. calabaricus lacks PGCs but shows granular leukocytes and a different type of NEC. Interestingly, ciliated cells and secretory cells show a dual morphology in E. calabaricus indicating the presence of cellular subtypes and suggesting more complex secretory activity. Also in E. calabaricus , cilia show a novel doublet‐membrane interaction that may control the displacement of the microtubule doublets. The subepithelium is a connective layer that appears thicker in P. senegalus and contains, in the two species, fibroblasts and granulocytes. The outer layer contains bundles of richly innervated striated muscle. This layer is likely involved in the control of lung motion. In the two species, smooth muscle cells constitute a limiting layer between the subepithelium and the striated muscle compartment. The role of this layer is unclear.     

Vertical displacement and taphonomic filtering of nannofossils by bioturbation in the Cretaceous–Palaeogene boundary section at Caravaca,SE Spain

K?dzierski, M., Rodríguez‐Tovar, F.J. & Uchman, A. 2011: Vertical displacement and taphonomic filtering of nannofossils by bioturbation in the Cretaceous–Palaeogene boundary section at Caravaca, SE Spain. Lethaia, Vol. 44, pp. 321–328. At the Caravaca section, SE Spain, the position of the Cretaceous–Palaeogene (K–Pg) boundary is well‐defined, with multidisciplinary datasets from a thin rusty layer at the base of the 10 cm dark boundary layer. Nannoplankton assemblages containing the Danian taxon Neochiastozygus sp. and enriched in Thoracosphaera spp. are displaced below the K–Pg boundary into the trace fossils Zoophycos, Thalassinoides and Chondrites. These trace fossils are filled with dark‐coloured sediments of the dark boundary layer. The nannofossil assemblage from the 1‐cm thick interval below the boundary, enriched in Thoracosphaera spp. and Braarudosphaera spp., may have been displaced by Chondrites tracemakers, the traces of which are abundant in this interval. The downward transport of the Danian nannofossils into the Maastrichtian by the tracemakers seems to be one of the common mechanisms responsible for their apparent appearance below the K–Pg boundary. The dark boundary layer contains very rare Danian specimens and abundant Maastrichtian nannofossils. The Maastrichtian taxa were most likely conveyed up on to the seafloor by tracemakers during the Danian. The redistribution of nannoplankton down and up across the rusty layer (K–Pg), challenges the usefulness of nannofossils for high‐resolution stratigraphy of the K–Pg boundary. □Nannoplankton, Cretaceous–Palaeogene boundary event, biogenic mixing, trace fossils.     

Characterization of uropathogenic E. coli O25b-B2-ST131, O15:K52:H1, and CGA: Neutrophils apoptosis,serum bactericidal assay,biofilm formation,and virulence typing

Pathogenic and drug-resistant strains of Escherichia coli (E. coli) O25b-B2-ST131, O15:H1-D-ST393, and CGA (clonal group A) clonal groups have spread worldwide. This study aimed at determining E. coli epidemic clonal groups, their virulence factors, biofilm formation, neutrophils apoptosis, and antimicrobial resistance pattern of uropathogenic E. coli. A total of 95 CTX-M-1-producing E. coli clinical isolates were enrolled. E. coli O25b-B2-ST131, CGA, and O15:K52:H1 were identified by serotyping and phylogrouping and allele-specific polymerase chain reaction-based assay. Antibiotic susceptibility, biofilm formation, hemolysis, and human serum bactericidal assay were performed. Neutrophil apoptosis was assayed by flow cytometry. Nine E. coli clonal groups including six O25b-B2-ST131 strains, two CGA, and one O15:K52:H1-D-ST393 strains were detected. One O25b-B2-ST131 isolate was a strong biofilm-producer. Three ST131 isolates had type I fimbriae. Furthermore, all the CGA and O15:K52:H1 and three of ST131 isolates harbored the P fimbriae. The virulence genes ompT, fimH, and traT were detected among all the clonal groups. The apoptosis was induced by O25b-B2-ST131, CGA, and O15:K52:H1 E. coli. There was no significant difference regarding apoptosis induction among clonal groups. Furthermore, the presence of the cdt, usp, and vat genes was significantly associated with the apoptosis of neutrophils by O25b-B2-ST131, CGA, and O15:K52:H1-D-ST393 clonal groups.     

Ecology and morphological characterization of gametophyte and 'Chantransia' stages of Sirodotia huillensis (Batrachospermales, Rhodophyta) from a stream in central Mexico

The morphology and phenology of Sirodotia huillensis was evaluated seasonally in a central Mexican first‐order calcareous stream. Water temperature was constant (24–25°C) and pH circumneutral to alkaline (6.7–7.9), and calcium and sulfates were the dominant ions. The gametophyte stages were characterized by the presence of a distinctive mucilaginous layer, a marked difference in phycocyanin to phycoerythrin ratio between female and male plants, and the presence of a carpogonia with a large trichogyne (>60 µm). Occasionally three capogonia were observed on a single basal cell. The ‘Chantransia’ stages were morphologically similar to those described for the other members of Batrachospermales. A remarkable observation was the formation of dome‐shaped structures, consisting of prostrate filaments that are related with the development of new gametophytes. Chromosome numbers were n = 4 for fascicle cells, cortical filament cells and dome‐shaped cells, and 2n = 8 for gonimoblast filament cells and ‘Chantransia’ stage filaments. Gametophytes and ‘Chantransia’ stages occurred in fast current velocities (60–170 cm/s) and shaded (33.1–121 µmol photons/m²/s) stream segments. The population fluctuated throughout the study period in terms of percentage cover and frequency: the ‘Chantransia’ stages were most abundant in the rainy season, whereas gametophytic plants had the highest frequency values during the dry season. These results were most likely a result of fluctuations in rainfall and related changes in current velocity. Some characteristics of this population can be viewed as probable adaptations to high current velocities: the mucilaginous layer around plants that reduces drag; potential increase in fertilization by the elongate and plentiful trichogynes and abundant dome‐shaped structures producing several gametophytes.     

Acetylation targets HSD17B4 for degradation via the CMA pathway in response to estrone

Dysregulation of hormone metabolism is implicated in human breast cancer. 17β-hydroxysteroid dehydrogenase type 4 (HSD17B4) catalyzes the conversion of estradiol (E2) to estrone (E1), and is associated with the pathogenesis and development of various cancers. Here we show that E1 upregulates HSD17B4 acetylation at lysine 669 (K669) and thereby promotes HSD17B4 degradation via chaperone-mediated autophagy (CMA), while a single mutation at K669 reverses the degradation and confers migratory and invasive properties to MCF7 cells upon E1 treatment. CREBBP and SIRT3 dynamically control K669 acetylation level of HSD17B4 in response to E1. More importantly, K669 acetylation is inversely correlated with HSD17B4 in human breast cancer tissues. Our study reveals a crosstalk between acetylation and CMA degradation in HSD17B4 regulation, and a critical role of the regulation in the malignant progression of breast cancer.     

Metamorphic change in EP37 expression: members of the βγ-crystallin superfamily in newt

 EP37 is an epidermis-specific protein found in the developing embryo of the Japanese newt, Cynops pyrrhogaster. Our previous study predicted the presence of genes homologous to EP37, which show temporary shared expression at the turn of metamorphosis. In this study, we isolated and characterized three cDNAs encoding novel EP37 homologues; two from the skin of an adult newt and the other from swimming larva. Conceptual translation of the open reading frames of these cDNAs predicted proteins carrying βγ-crystallin motifs and putative calcium-binding sites, both of which are features shared by the originally identified EP37 (EP37L1), as well as a spore coat protein of Myxococcus xanthus, protein S. Immunoblot analyses and immunohistochemical studies indicated that two of the EP37 proteins, EP37L1 and EP37L2, are exclusively expressed in the epidermis (skein cells) including the figures of Eberth at premetamorphic stages. During and after metamorphosis, the expression of EP37 proteins was mainly observed in cutaneous glands, and a molecular transition to the adult types of EP37, EP37A1 and EP37A2, occurred. These observations suggest that EP37 proteins play an important role in construction of integumental tissues and adaptation to the aquatic or amphibious environment. Received: 6 September 1996 / Accepted: 30 October 1996     

Taxonomic value of seed characters in the Erica tetralix L. group (Ericaceae)

A study of the seed morphology of Erica tetralix and the related species E. mackaiana and E. andevalensis was performed to evaluate the taxonomic importance of seed characters. Size and shape of seeds and surface cells were studied, and area, perimeter, main axis lengths, elongation and sinuosity were measured. Levels of intraspecific variation were considered by means of nested analysis of variance. The study of the intraspecific variation of the characters shows significant differences for lower levels, higher than variations between species for most characters. Levels of variation must be based upon the numerical analysis of seed characters for a correct approach to determining systematic value. E. mackaiana and E. andevalensis group together versus E. tetralix for sinuosity of testa cells. This is a good taxonomic character because it shows significant differences between species and no significant differences between populations. E. tetralix has strongly undulated testa cells while E. mackaiana and E. andevalensis have slightly undulated or curved testa cells. These two species can be discriminated by size of the seed, with seeds of 0.37-0.38 mm for E. andevalensis and 0.4 - 0.45 mm for E. mackaiana. Similarities are found in seed morphology for E. mackaiana and E. andevalensis, but we nevertheless consider the species status for the three taxa as the most natural taxonomic assignment. A key to the species is provided.     

N‐acetylcysteine increases susceptibility of HeLa cells to bacterial invasion

Serratia grimesii are non‐pathogenic bacteria capable, however, to invade eukaryotic cells provided that they synthesize intracellular metalloprotease grimelysin (Bozhokina et al. [2011] Cell. Biol. Int. 35: 111–118). To elucidate how invasion of grimelysin containing bacteria depends on physiological state of host cells, we studied the effect of N‐acetylcysteine (NAC) on susceptibility of HeLa cells to invasion by the wild‐type S. grimesii and recombinant E. coli expressing grimelysin gene. Incubation of HeLa cells with 10 mM NAC resulted in changes of cell morphology and disassembly of actin cytoskeleton that were reversed when NAC was removed from the culture medium. Both in the presence of NAC and upon its removal, the entry of grimelysin producing bacteria increased by a factor of 1.5–2 and 3–3.5 for wild‐type S. grimesii and recombinant E. coli, respectively. This effect does not correlate with cytoskeleton rearrangements but may be due to the NAC‐induced up‐regulation of cell surface receptors playing a role in cell adhesion and cell–cell junctions. A twofold difference in the efficiency of S. grimesii and recombinant E. coli to enter the NAC‐treated cells suggests that the entry of the wild‐type and recombinant bacteria occurs via different receptors which activity is differently affected by NAC. J. Cell. Biochem. 114: 1568–1574, 2013. © 2013 Wiley Periodicals, Inc.