Improving the Safety of Staphylococcus aureus Polyvalent Phages by Their Production on a Staphylococcus xylosus Strain

Team1 (vB_SauM_Team1) is a polyvalent staphylococcal phage belonging to the Myoviridae family. Phage Team1 was propagated on a Staphylococcus aureus strain and a non-pathogenic Staphylococcus xylosus strain used in industrial meat fermentation. The two Team1 preparations were compared with respect to their microbiological and genomic properties. The burst sizes, latent periods, and host ranges of the two derivatives were identical as were their genome sequences. Phage Team1 has 140,903 bp of double stranded DNA encoding for 217 open reading frames and 4 tRNAs. Comparative genomic analysis revealed similarities to staphylococcal phages ISP (97%) and G1 (97%). The host range of Team1 was compared to the well-known polyvalent staphylococcal phages phi812 and K using a panel of 57 S. aureus strains collected from various sources. These bacterial strains were found to represent 18 sequence types (MLST) and 14 clonal complexes (eBURST). Altogether, the three phages propagated on S. xylosus lysed 52 out of 57 distinct strains of S. aureus. The identification of phage-insensitive strains underlines the importance of designing phage cocktails with broadly varying and overlapping host ranges. Taken altogether, our study suggests that some staphylococcal phages can be propagated on food-grade bacteria for biocontrol and safety purposes.     

Protein expression during seed development in Araucaria angustifolia: transient accumulation of class IV chitinases and arabinogalactan proteins

Protein accumulation and patterns during embryogenesis in the recalcitrant seeds of the gymnosperm species Araucaria angustifolia (Bert.) O. Kuntze were studied. Soluble seed proteins, chitinases, and arabinogalactan proteins (AGPs) were analyzed by means of 2-D gel electrophoresis, mass spectrometry, isoelectric focusing, Western blot, precipitation and staining with β-glucosyl Yariv reagent (β-Glc)₃Y, and gas liquid chromatography. Despite the recalcitrant nature of the seeds, the electrophoretic patterns of A . angustifolia seed proteins showed similarities with orthodox seed types. Proteins showing chitinolytic activity were observed in all seed stages analyzed, but the expression of class IV chitinases was restricted to late stages of seed development. AGPs were prominent during stages of seed development characterized by intensive cell division and differentiation, and their decrease during seed maturation might be related to cell wall modifications during the deposition of storage compounds. Gas liquid chromatographic analyzes of AGPs did not show quantitative changes in their carbohydrate moieties during seed development. A low molecular weight protein specifically expressed in mature seeds was precipitated using (β-Glc)₃Y. Amino acid sequences obtained from MS/MS analysis revealed peptides rich in valine and acidic amino acids, but devoid in amino acids normally found in AGPs polypeptides, suggesting that these peptides are not related to classical or non-classical AGPs. Possible implications of chitinases and AGPs during seed development in A . angustifolia are discussed.     

Non-Newtonian behaviour of yeast suspensions

Summary The rheological properties of yeast (Baker's yeast) suspensions in water and in water-methanol-electrolyte solutions have been investigated using a Fann V.G. concentric cylinder viscometer. The resulted rheograms could be described by the power law model for non-Newtonian fluids with the flow behaviour index n<1 indicating pseudo-plastic behaviour. This flow behaviour index was constant under all operating conditions while the consistancy index k increased exponentially with increasing wet yeast volume fraction. The addition of methanol and electrolytes had a negligible effect on the rheological properties. Changing the pH of the suspension acidic to lowered the apparent viscosities.     

Fine needle aspiration cytology of well-differentiated papillary mesothelioma: a case report

BACKGROUND: Well-differentiated papillary mesothelioma (WDPM) is an uncommon subtype of mesothelioma that typically occurs in the peritoneum of women without a history of asbestos exposure and usually follows an indolent clinical course. Fine needle aspiration (FNA) of this type of tumor has rarely been reported. CASE: A 64-year-old woman with 11-year history of colon cancer and an adrenal nodule was found, on abdominal computed tomography, to have a mass in the right lobe of the liver. Aspirates of the mass were composed of abundant, tight, papillary groups, monolayered, pavementlike sheets; and scattered single cells with minimal atypia. The cell block showed a predominantly papillary growth pattern and a single layer of bland, cuboidal to flattened covering cells with stout, fibrovascular cores containing clusters of foamy histiocytes. Tumor cells in the focal tubulopapillary and solid areas were mingled with inflammatory cells and showed slightly more atypia than did the cells covering the papillae. The differential diagnoses were intrahepatic papillary neoplasm, including well-differentiated mesothelioma and metastatic low grade papillary serous carcinoma. At surgery the tumor was found to be a pedunculated peritoneal mass that arose from the posterior surface of the right lobe of the liver. The mesothelial origin of the tumor was confirmed by both immunoperoxidase study and electron microscopic examination, which demonstrated long, slender, branching microvilli. CONCLUSION: Familiarity with the cytomorphologic features and clinical presentation of WDPM, knowledge of the exact anatomic location and consideration of the appropriate differential diagnosis combined with ancillary studies are the keys to an accurate diagnosis.     

Acrylonitrile interaction with testicular DNA in rats

In the present study we report the in vivo interaction of acrylonitrile (VCN) with testicular tissue in rats. Covalent binding of radioactivity to testicular tissue DNA was examined for a period of 72 hr after a single oral dose (46.5 mg/kg) of [2, 3-¹⁴C] VCN. Maximal covalent binding was observed at 0.5 hr (8.9 μmol VCN equivalent/mol nucleotide). Binding decreased gradually thereafter but was still detected (2.5 μmol VCN equivalent/mol nucleotide) at 72 hr following VCN administration. Further, we examined the effects of VCN on DNA synthesis and repair in the testes of rats following a single oral dose (46.5 mg/kg) of VCN to clarify the impact of the covalent binding observed on the testicular genetic material. A significant decrease in DNA synthesis (80% of control) was observed at 0.5 hr after treatment. At 24 hr following acrylonitrile administration, testicular DNA synthesis was severely inhibited (38% of control). Testicular DNA repair was increased 1.5-fold at 0.5 hr and more than 3.3-fold at 24 hr following treatment with VCN. These results suggest that VCN can act as a multipotent genotoxic agent by alkylating DNA in testicular tissue and may affect the male reproductive function by interfering with testicular DNA synthesis and repair processes.     

Detection of phytochrome-like genes from Rhazya stricta (Apocynaceae) using de novo genome assembly

Phytochrome-like genes in the wild plant species Rhazya stricta Decne were characterized using a de novo genome assembly of next generation sequence data. Rhazya stricta contains more than 100 alkaloids with multiple pharmacological properties, and leaf extracts have been used to cure chronic rheumatism, to treat tumors, and in the treatment of several other diseases. Phytochromes are known to be involved in the light-regulated biosynthesis of some alkaloids. Phytochromes are soluble chromoproteins that function in the absorption of red and far-red light and the transduction of intracellular signals during light-regulated plant development. De novo assembly of the nuclear genome of R. stricta recovered 45,641 contigs greater than 1000 bp long, which were used in constructing a local database. Five sequences belonging to Arabidopsis thaliana phytochrome gene family (i.e., AtphyABCDE) were used to identify R. stricta contigs with phytochrome-like sequences using BLAST. This led to the identification of three contigs with phytochrome-like sequences covering AtphyA-, AtphyC- and AtphyE-like full-length genes. Annotation of the three sequences showed that each contig consists of one phytochrome-like gene with three exons and two introns. BLASTn and BLASTp results indicated that RsphyA mRNA and protein sequences had homologues in Wrightia coccinea and and Solanum tuberosum, respectively. RsphyC-like mRNA and protein sequence were homologous to Vitis vinifera and Vitis riparia. RsphyE-like mRNA coding and protein sequences were homologous to Ipomoea nil. Multiple-sequence alignment of phytochrome proteins indicated a homology with 30 sequences from 23 different species of flowering plants. Phylogenetic analysis confirmed that each R. stricta phytochrome gene is related to the same phytochrome gene of other flowering plants. It is proposed that the absence of phyB gene in R. stricta is due to RsphyA gene taking over the role of phyB.     

Catabolism of itaconic acid by preformed mats of aspergillus terreus

Summary The breakdown of itaconic acid by preformed mats of a local strain of Aspergillus terreus was studied using the zone-strip technique. Low p_H values restrained the uptake of itaconic acid and the accumulation of various other acids while higher p_H values exerted an opposite effect. The results obtained when using various enzyme inhibitors were those anticipated on basis of the direct transformation of itaconic to aconitic acid through the fixation of CO₂ and of the existence of the T.A.C. (tricarboxylic acid cycle) as a main metabolic channel operating in this organism.