Novel Inhibitor of E. coli DNA Adenine Methyltransferase (Ecodam)

EcoDam is an adenine-N6 DNA methyltransferase that methylates the GATC sites in the Escherichia coli genome. DNA-adenine methylation is not present in higher eukaryotes including humans. These observations raise the possibility that dam inhibitors may be used as anti-microbial agents. Polyphosphate (Poly(P)) is an important metabolite and signaling molecule in prokaryotes and eukaryotes. Here, by using gel retardation experiments to investigate the competition of DNA binding by EcoDam in the presence of polyphosphate, we found that Poly (P) strongly interferes with DNA binding by EcoDam, while same concentration of monophosphate does not. In addition, we demonstrated that Poly (P) binding inhibits the activity of EcoDam and our results suggest that Poly (P) led to strong inhibition of the EcoDam catalytic activity, while monophosphate had only moderate effect.

X-ray Crystallographic Analysis of MinC N-Terminal Domain from Escherichia coli

MinC plays an important role in bacterial cell division system by inhibiting FtsZ assembly. However, the molecular mechanism of the action is poorly understood. E. coli MinC Nterminus domain was purified and crystallized using 1.4 M sodium citrate pH 6.5 as a precipitant. X-ray diffraction data was collected and processed to 2.3 Å from a native crystal. The crystal belonged to space group P212121, with the unit cell parameters a = 52.7, b = 54.0, c = 64.7 Å. Assuming the presence of two molecules in the asymmetric unit, the Matthews coefficient value is 1.94 Å3 Da-1, which corresponds to a solvent content of 36.5%. The overall structure of MinCN is observed as a dimer form through anti-parallel ß-strand interaction.

Study on Phytochemical Properties, Antibacterial Activity and Cytotoxicity of Aloe vera L.

The aim of the study was to investigate phytochemical properties, antimicrobial activity and cytotoxicity of Aloe vera. The phytochemical screening of the extracts of leaves of A. vera revealed the presence of bioactive compounds such as alkaloids, tannins, flavonoids phenolic compounds, and etc. with absence of cyanogenic glycosides. Three different solvents such as methanol, ethanol and Di-Methyl sulfoxide were used to screen the antimicrobial activity of A. vera leaves against four human clinical pathogens by agar well diffusion method. The maximum antibacterial activities were observed in methanol extract followed by ethanol and Di-Methyl sulfoxide. It was also found that remarkable antibacterial activities with methanolic and ethanolic extracts of A. vera compared with the standard antibiotic, tetracycline that was not active against E. coli and S. boydii and supported the view that A. vera is a potent antimicrobial agent compared with the conventional antibiotic. Moreover, the brine shrimps (Artemia salina) toxicity test exhibited LC50 value was 569.52 ppm. The resulting data indicated that the A. vera plant have less toxic effects on brine shrimp. Hence, it is signified that Aloe vera plant extract is safe to be used as an antimicrobial agent.

Incidence of Pathogenic Bacteria in Cakes and Tarts Displayed for Sale in Tripoli, Libya

This study was conducted to investigate the incidence of pathogenic bacteria: Salmonella, Shigella, Escherichia coli O157 and Staphylococcus aureus in cakes and tarts collected from thirtyfive confectionery producing and selling premises located within Tripoli city, Libya. The results revealed an incidence of S. aureus with 94.4 and 48.0 %, E. coli O157 with 14.7 and 4.0 % and Salmonella sp. with 5.9 and 8.0 % in cakes and tarts samples respectively; while Shigella was not detected in all samples. In order to determine the source of these pathogenic bacteria, cotton swabs were taken from the hands of workers on the production line, the surfaces of preparation tables and cream whipping instruments. The results showed that the cotton swabs obtained from the hands of workers contained S. aureus and Salmonella sp. with an incidence of 42.9 and 2.9 %, the cotton swabs obtained from the surfaces of preparation tables 22.9 and 2.9 % and the cotton swabs obtained from the cream whipping instruments 14.3 and 0.0 % respectively; while E. coli O157 and Shigella sp. were not detected in all swabs. Additionally, other bacteria were isolated from the hands of workers and the Surfaces of producing equipments included: Aeromonas sp., Pseudomonas sp., E. coli, Klebsiella sp., Enterobacter sp., Citrobacter sp., Proteus sp., Serratia sp. and Acinetobacter sp. These results indicate that some of the cakes and tarts might pose threat to consumer's health. Meanwhile, occurrences of pathogenic bacteria on the hands of those who are working in production line and the surfaces of equipments reflect poor hygienic practices at most confectionery premises examined in this study. Thus, firm and continuous surveillance of these premises is needed to insure the consumer's health and safety.

Ecosystems of Lake Sevan Basin-s Rivers in Armenia

Taking into account the importance of Lake Sevan and Lake Sevan basin-s rivers for Armenian economy, the main goals of our investigations were the documentation of water quality and the biodiversity of invertebrates developed in Lake Sevan basin-s rivers and selected tributaries. Moderately satisfied ecological condition for the biodiversity of Lake Sevan basin-s rivers has been established, and the changes in species- composition of zoobenthos in Lake Sevan were detected. A growing tendency of antibiotic resistance among E. coli isolates in water resources has been shown.

Cloning and Expression of D-Threonine Aldolase from Ensifer arboris NBRC100383

D-erythro-cyclohexylserine (D chiral unnatural β-hydroxy amino acid expected for the synthesis of drug for AIDS treatment. To develop a continuous bioconversion system with whole cell biocatalyst of D-threonine aldolase (D genes for the D-erythro-CHS production, D-threonine aldolase gene was amplified from Ensifer arboris 100383 by direct PCR amplication using two degenerated oligonucleotide primers designed based on genomic sequence of Shinorhizobium meliloti Sequence analysis of the cloned DNA fragment revealed one open-reading frame of 1059 bp and 386 amino acids. This putative D-TA gene was cloned into NdeI and EcoRI (pEnsi His-tag sequence or BamHI (pEnsi-DTA[2]) sequence of the pET21(a) vector. The expression level of the cloned gene was extremely overexpressed by E. coli BL21(DE3) transformed with pEnsi-DTA[1] compared to E. coli BL21(DE3) transformed with pEnsi-DTA[2]. When the cells expressing the wild used for D-TA enzyme activity, 12 mM glycine was successfully detected in HPLC analysis. Moreover, the whole cells harbouring the recombinant D-TA was able to synthesize D-erythro of 0.6 mg/ml in a batch reaction.

Prevention of Biofilm Formation in Urinary Catheter by Coating Enzymes/ Gentamycin/ EDTA

Urinary Tract Infections (UTI) account for an estimated 25-40% nosocomial infection, out of which 90% are associated with urinary catheter, called Catheter associated urinary tract infection (CAUTI). The microbial populations within CAUTI frequently develop as biofilms. In the present study, microbial contamination of indwelling urinary catheters was investigated. Biofilm forming ability of the isolates was determined by tissue culture plate method. Prevention of biofilm formation in the urinary catheter by Pseudomonas aeruginosa was also determined by coating the catheter with some enzymes, gentamycin and EDTA. It was found that 64% of the urinary catheters get contaminated during the course of catheterization. Of the total 6 isolates, biofilm formation was seen in 100% Pseudomonas aeruginosa and E. coli, 90% in Enterococci, 80% in Klebsiella and 66% in S. aureus. It was noted that the biofilm production by Pseudomonas was prolonged by 7 days in amylase, 8 days in protease, 6 days in lysozyme, 7days in gentamycin and 5 days in EDTA treated catheter.

Early Onset Neonatal Sepsis Pathogens in Malaysian Hospitals: Determining Empiric Antibiotic

Information regarding early onset neonatal sepsis (EONS) pathogens may vary between regions. Global perspectives showed Group B Streptococcal (GBS) as the most common causative pathogens, but the widespread use of intrapartum antibiotics has changed the pathogens pattern towards gram negative microorganisms, especially E. coli. Objective of this study is to describe the pathogens isolated, to assess current treatment and risk of EONS. Records of 899 neonates born in three General Hospitals between 2009 until 2012 were retrospectively reviewed. Proven was found in 22 (3%) neonates. The majority was isolated with gram positive organisms, 17 (2.3%). All grams positive and most gram negative organisms showed sensitivity to the tested antibiotics. Only two rare gram negative organisms showed total resistant. Male was possible risk of proven EONS. Although proven EONS remains uncommon in Malaysia, nonetheless, the effect of intrapartum antibiotics still required continuous surveillance.