Acinetobacter baumannii has emerged as an increasing multidrug-resistant threat in hospitals and a common opportunistic nosocomial pathogen worldwide. However, molecular details of the pathogenesis and physiology of this bacterium largely remain to be elucidated. Here we identify and characterize the c-di-GMP signalling network and assess its role in biofilm formation and surface associated motility. Bioinformatic analysis revealed eleven candidate genes for c-di-GMP metabolizing proteins (GGDEF/EAL domain proteins) in the genome of A. baumannii strain 17978. Enzymatic activity of the encoded proteins was assessed by molecular cloning and expression in the model organisms Salmonella typhimurium and Vibrio cholerae. Ten of the eleven GGDEF/EAL proteins altered the rdar morphotype of S. typhimurium and the rugose morphotype of V. cholerae. The over expression of three GGDEF proteins exerted a pronounced effect on colony formation of A. baumannii on congo Red agar plates. Distinct panels of GGDEF/EAL proteins were found to alter biofilm formation and surface associated motility of A. baumannii upon over expression. The GGDEF protein A1S_3296 appeared as a major diguanylate cyclase regulating macro-colony formation, biofilm formation and the surface associated motility. AIS_3296 promotes Csu pili mediated biofilm formation. We conclude that a functional c-di-GMP signalling network in A. baumannii regulates biofilm formation and surface associated motility of this increasingly important opportunistic bacterial pathogen.

Background: Acinetobacter baumannii is a Gram-negative opportunistic pathogen with a notorious reputation of being resistant to antimicrobial agents. The capability of A. baumannii to persist and disseminate between healthcare settings has raised a major concern worldwide. Methods: Our study investigated the antibiotic resistance features and molecular epidemiology of 52 clinical isolates of A. baumannii collected in Pakistan between 2013 and 2015. Antimicrobial susceptibility patterns were determined by the agar disc diffusion method. Comparative sequence analyses of the ampC and bla OXA-51-like alleles were used to assign the isolates into clusters. The whole genomes of 25 representative isolates were sequenced using the MiSeq Desktop Sequencer. Free online applications were used to determine the phylogeny of genomic sequences, retrieve the multilocus sequence types (ST), and detect acquired antimicrobial resistance genes.Results: Overall, the isolates were grouped into 7 clusters and 3 sporadic isolates. The largest cluster, Ab-Pak-cluster-1 (bla OXA-66 and ISAba1-ampC-19) included 24 isolates, belonged to ST2 and International clone (IC) II, and was distributed between two geographical far-off cities, Lahore and Peshawar. Ab-Pak-clusters-2 (bla OXA-66 , ISAba1-ampC-2), and -3 (bla OXA-66 , ISAba1-ampC-20) and the individual isolate Ab-Pak-Lah-01 (ISAba1-bla OXA-66 , ISAba1-ampC-2) were also assigned to ST2 and IC II. On the other hand, Ab-Pak-clusters-4 (bla OXA-69 , ampC-1), -5 (bla OXA-69 , ISAba1-ampC-78), and -6A (bla OXA-371 , ISAba1-ampC-3) belonged to ST1, while Ab-Pak-cluster-6B (bla OXA-371 , ISAba1-ampC-8) belonged to ST1106, with both ST1 and ST1106 being members of IC I. Five isolates belonged to Ab-Pak-cluster-7 (bla . This cluster corresponded to ST158, showed a well-delineated position on the genomic phylogenetic tree, and was equipped with several antimicrobial resistance genes including bla OXA-23 and bla . Conclusions:Our study detected the occurrence of 7 clusters of A. baumannii in Pakistan. Altogether, 6/7 of the clusters and 45/52 (86.5%) of the isolates belonged to IC I (n = 9) or II (n = 36), making Pakistan no exception to the global domination of these two clones. The onset of ST158 in Pakistan marked a geographical dispersal of this clone beyond the Middle East and brought up the need for a detailed characterization.

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Objective: The aim of study was to assess hospital phobia and its variation of depressive symptoms in primary care population. Introduction: Nosocomephobia is the fear of hospitals. It is a fairly common phobia; many people are known to suffer from it. Like that “If I go to a hospital, I’m fairly sure it is a fact that my life is not guaranteed.” Patients are afraid of hospitals, especially the emergency rooms. Hospitals are the mark of cure and health. They even usually result in huge expenses. Most people understand that it is a medical necessity and that one does not and fear must be overcome in case of Nosocomephobia though, the patient simply refuses to go to a hospital and the result often are not desirable including death especially in case of major life threatening conditions. Results: In Primary Care population (PCP) severity of Hospital-phobia is very high. Patients show different depressive symptoms like nausea, loss of appetite, mood swing etc. The result of survey shows that insomnia is the highest number found in depressive patients which is approximately 50% in male patients alone. Female the percentage is slightly low i.e. approximately 40%. Discussion and Conclusion: A survey has been conducted at Karachi to ascertain how many people are suffering from hospital phobia. Our survey result showed that the hospital fear (nosocomephobia) is very common in the biggest city of Pakistan i.e. Karachi. The best way to overcome the nosocomephobia is to change the mindset of the patient through some self-help techniques.

Objectives: To evaluate the activity of Methylglyoxal against the blood culture isolates of Salmonella Typhi and various Gram negative rods and to compare the activity of Methylglyoxal against S. Typhi and other Gram negative rods. Methods: It was an experimental study conducted at the Department of Microbiology, University of Health Sciences (UHS), Lahore-Pakistan in collaboration with the Department of Microbiology, CMH Lahore, from July 2011 to June 2012. Recent blood culture isolates of S. Typhi and other Gram negative rods were collected from different hospitals of Lahore and kept stored at -80°C. As per the latest CLSI guidelines, morphological, biochemical and serological identification was carried out and antimicrobial susceptibility was tested. A multi-point inoculator was used to carry out agar dilution for determination of MICs of MGO. Results were determined after compilation of data using latest SPSS version. Results: MIC90 of MGO against the clinical isolates of S. Typhi was 0.20 mg/mL (2.8 mM) and against Gram negative rods it was 0.21 mg/mL (3.0 mM). The p-value of MICs of MGO against the isolates of S. Typhi was 0.023 when compared with Gram negative rods (p<0.05; statistically significant). Conclusion: MGO has a scientifically proven in vitro antimicrobial activity against blood culture isolates of S. Typhi and various Gram negative rods. doi: https://doi.org/10.12669/pjms.35.4.807 How to cite this:Afzal RK, Khalid F, Hannan A, Ahmed SA. Methylglyoxal: Antimicrobial activity against blood culture isolates of Salmonella Typhi and other Gram negative rods. Pak J Med Sci. 2019;35(4):---------. doi: https://doi.org/10.12669/pjms.35.4.807 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.