YY1 is a transcriptional activator of mouse LINE-1 Tf subfamily.

Long interspersed element type 1 (LINE-1, L1) is an active autonomous transposable element (TE) in the human genome. The first step of L1 replication is transcription, which is controlled by an internal RNA polymerase II promoter in the 5' untranslated region (UTR) of a full-length L1. It has been shown that transcription factor YY1 binds to a conserved sequence motif at the 5' end of the human L1 5'UTR and dictates where transcription initiates but not the level of transcription. Putative YY1-binding motifs have been predicted in the 5'UTRs of two distinct mouse L1 subfamilies, Tf and Gf. Using site-directed mutagenesis, in vitro binding, and gene knockdown assays, we experimentally tested the role of YY1 in mouse L1 transcription. Our results indicate that Tf, but not Gf subfamily, harbors functional YY1-binding sites in its 5'UTR monomers. In contrast to its role in human L1, YY1 functions as a transcriptional activator for the mouse Tf subfamily. Furthermore, YY1-binding motifs are solely responsible for the synergistic interaction between monomers, consistent with a model wherein distant monomers act as enhancers for mouse L1 transcription. The abundance of YY1-binding sites in Tf elements also raise important implications for gene regulation at the genomic level.

Subfamily-specific differential contribution of individual monomers and the tether sequence to mouse L1 promoter activity.

BACKGROUND: The internal promoter in L1 5'UTR is critical for autonomous L1 transcription and initiating retrotransposition. Unlike the human genome, which features one contemporarily active subfamily, four subfamilies (A_I, Gf_I and Tf_I/II) have been amplifying in the mouse genome in the last one million years. Moreover, mouse L1 5'UTRs are organized into tandem repeats called monomers, which are separated from ORF1 by a tether domain. In this study, we aim to compare promoter activities across young mouse L1 subfamilies and investigate the contribution of individual monomers and the tether sequence. RESULTS: We observed an inverse relationship between subfamily age and the average number of monomers among evolutionarily young mouse L1 subfamilies. The youngest subgroup (A_I and Tf_I/II) on average carry 3-4 monomers in the 5'UTR. Using a single-vector dual-luciferase reporter assay, we compared promoter activities across six L1 subfamilies (A_I/II, Gf_I and Tf_I/II/III) and established their antisense promoter activities in a mouse embryonic fibroblast cell line and a mouse embryonal carcinoma cell line. Using consensus promoter sequences for three subfamilies (A_I, Gf_I and Tf_I), we dissected the differential roles of individual monomers and the tether domain in L1 promoter activity. We validated that, across multiple subfamilies, the second monomer consistently enhances the overall promoter activity. For individual promoter components, monomer 2 is consistently more active than the corresponding monomer 1 and/or the tether for each subfamily. Importantly, we revealed intricate interactions between monomer 2, monomer 1 and tether domains in a subfamily-specific manner. Furthermore, using three-monomer 5'UTRs, we established a complex nonlinear relationship between the length of the outmost monomer and the overall promoter activity. CONCLUSIONS: The laboratory mouse is an important mammalian model system for human diseases as well as L1 biology. Our study extends previous findings and represents an important step toward a better understanding of the molecular mechanism controlling mouse L1 transcription as well as L1's impact on development and disease.

Isolation and characterisation of carbapenem-resistant Pseudomonas aeruginosa from hospital environments in tertiary care hospitals in Dhaka, Bangladesh.

OBJECTIVES: Increasing evidence of carbapenem-resistant Pseudomonas aeruginosa (CRPA) infection in healthcare facilities poses an alarming threat to public health. There is little evidence on the occurrence of this organism in Bangladeshi hospitals. METHODS: We collected 117 environmental swab samples from two tertiary care hospitals in Dhaka, Bangladesh and tested for Pseudomonas species by nonselective enrichment of swabs followed by plating on Cetrimide agar. We confirmed the isolates as P. aeruginosa by API 20NE test and polymerase chain reaction Polymerase Chain Reaction (PCR) for 16S rRNA gene. We analysed P. aeruginosa isolates for susceptibility against 15 clinically important antibiotics and tested the carbapenem-resistant isolates for metallo ß-lactamase (MBL). All CRPA isolates were characterised for carbapenem-resistant genes, virulence genes and biofilm formation genes. RESULTS: Of 117 swab samples, 82 (70%) were tested positive for P. aeruginosa. All P. aeruginosa isolates were multidrug-resistant, and 39% (n = 32) of isolates were CRPA. Around 56% (n = 18) of CRPA were MBL-producing; 22% (n = 7) of isolates were positive for carbapenemase gene blaNDM followed by 16% (n = 5) for blaVIM and 13% (n = 4) for blaIMP. Sequencing identified these genes as blaNDM-1, blaIMP-13, blaVIM-2 variants. Based on optical density values, 94% (n = 30) of CRPA isolates were capable of producing biofilms. All CRPA isolates (n = 32) were positive for at least 1 of 6 biofilm-associated genes and 4 of 12 virulence genes tested in the study. CONCLUSION: Hospital environments in Bangladesh are contaminated with highly virulent CRPA, which might be a potential source of hospital-acquired infections, accentuating the need for strengthening hospital infection control programs.

Antibacterial activities of green tea crude extracts and synergistic effects of epigallocatechingallate (EGCG) with gentamicin against MDR pathogens.

Plant extracts and their purified compounds were examined for synergistic antimicrobial activity using selected multi-drug resistant (MDR) pathogens. The study aims to investigate the antibacterial activity of green tea (Camellia sinensis) and its purified compound epigallocatechingallate (EGCG). The synergistic relation of the compound with antibiotic was detected against selected potential Gram positive and Gram negative pathogens. Staphylococcus aureus and Escherichia coli were used as test pathogens which were resistant to different groups of antibiotics. After collection of fresh green tea leaves, samples were washed and air dried. EGCG is one of the bioactive compounds and was separated from tea plant. Antibacterial activity of EGCG and crude extracts of green tea were done by microdilution method (minimum inhibitory concentration and minimum bactericidal concentration). The synergistic effect of EGCG and gentamicin was determined. MIC value of green tea extract was found at 125 µg/mL in case of MDR E. coli, MDR S. aureus and their reference strains and MBC at 500 µg/mL against S. aureus. No MBC value was found against E. coli. EGCG showed better activity on Gram positive pathogen compared to that of Gram negative. MBC value of this compound was 1250 µg/mL for E. coli where 625 µg/mL for S. aureus. Strong synergistic relation (FICI 0.325) was found against pathogens in the combination of EGCG with gentamycin. The purified EGCG compound of green tea has great synergistic effect against MDR pathogens. More investigation is needed to know the inhibitory effect of these plant extracts and their components.

Bacteriological assessments of foodborne pathogens in poultry meat at different super shops in Dhaka, Bangladesh.

Poultry is now considered as a major fast-growing source of meat in the world. The consumers demand safe and hygienic products without contamination with pathogenic microorganisms when the production and consumption of poultry meat is gradually increasing. The present study was conducted to assess the bacterial contamination of dressed chicken collected from different supershops in Dhaka, Bangladesh. The chicken samples from S1, S2, M1, M2 and A supershops were analyzed to determine the enteropathogenic bacteria in poultry meat. Three genera of bacteria were isolated from all of the chicken meat samples. These enteropathogens from various organs of dressing chickens were also enumerated. The isolates were presumptively identified as E. coli, Salmonella spp., and Shigella spp. by conventional culture method. The three enteropathogens were subjected to PCR assay for their confirmation as virulent enteropathogens. Only E. coli isolates were confirmed as pathogenic E. coli (Enterotoxigenic), other isolates were not confirmed as virulent Salmonella spp., Shigella spp.. Results of this study demonstrated that more cautions are recommended for personnel hygiene in processing and handling of poultry and poultry products to prevent occurrence of enterotoxigenic E. coli in dressed poultry meat sold by the supershops in Bangladesh.