Elucidation of molecular mechanism for colistin resistance among Gram-negative isolates from tertiary care hospitals.

Antimicrobial resistance is a growing concern of global public health. The emergence of colistin-resistance among carbapenem-resistant (CPR) Gram-negative bacteria causing fear of pan-resistance, treatment failure, and high mortality across the globe. AIM: To determine the genotypic colistin-resistance mechanisms among colistin-resistant (CR)Gram-negative clinical isolates along with genomic insight into hypermucoviscous(hv)-CR-Klebsiella pneumoniae. METHODS: Phenotypic colistin-resistance via broth-microdilution method. PCR-based detection of plasmid-mediated colistin resistance genes(mcr-1,2,3). Characterization of selected hvCR-K. pneumoniae via Whole-genome sequencing. RESULTS: Phenotypic colistin-resistance was 28% among CPR-Gram-negative isolates of which 90% of CR-isolates displayed MDR profile with overall low plasmid-mediated colistin resistance (mcr-2 = 9.4%;mcr-3 = 6%). Although K. pneumoniae isolates showed the highest phenotypic colistin-resistance (51%) however, relatively low plasmid-mediated gene-carriage (mcr-2 = 11.5%;mcr-3 = 3.4%) pointed toward other mechanisms of colistin-resistance. mcr-negative CR-K. pneumoniae displaying hv-phenotype were subjected to WGS. In-silico analysis detected 7-novel mutations in lipid-A modification genes includes eptA(I38V; V50L; A135P), opgE(M53L; T486A; G236S), and arnD(S164P) in addition to several non-synonymous mutations in lipid-A modification genes conferring resistance to colistin. Insertion of 6.6-kb region harboring putative-PEA-encoding gene(yjgX) was detected for the first time in K. pneumoniae (hvCRKP4771). In-silico analysis further confirmed the acquisition of not only MDR determinants but several hypervirulent-determinants displaying a convergent phenotype. CONCLUSION: overall high prevalence of phenotypic colistin resistance but low mcr-gene carriage suggested complex chromosomal mediated resistance mechanism especially in K. pneumoniae isolates. The presence of novel mutations in lipid-A modification genes and the acquisition of putative-PEA-encoding gene by hvCR-K. pneumoniae points toward the role of chromosomal determinants conferring resistance to colistin in the absence of mcr-genes.

Shifting the balance between goals and habits: Five failures in experimental habit induction.

Habits are repetitive behaviors that become ingrained with practice, routine, and repetition. The more we repeat an action, the stronger our habits become. Behavioral and clinical neuroscientists have become increasingly interested in this topic because habits may contribute to aspects of maladaptive human behavior, such as compulsive behavior in psychiatry. Numerous studies have demonstrated that habits can be induced in otherwise healthy rats by simply overtraining stimulus-response behaviors. However, despite growing interest in this topic and its application to psychiatry, a similar body of work in humans is absent. Only a single study has been published in humans that shows the effect of extensive training on habit expression. Here, we report five failed attempts to demonstrate that overtraining instrumental behavior leads to the development of inflexible habits in humans, using variants of four previously published outcome devaluation paradigms. Extensive training did not lead to greater habits in two versions of an avoidance learning task, in an appetitive slips-of-action task, or in two independent attempts to replicate the original demonstration. The finding that these outcome devaluation procedures may be insensitive to duration of stimulus-response training in humans has implications for prior work in psychiatric populations. Specifically, it converges with the suggestion that the failures in outcome devaluation in compulsive individuals reflect dysfunction in goal-directed control, rather than overactive habit learning. We discuss why habits are difficult to experimentally induce in healthy humans, and the implications of this for future research in healthy and disordered populations. (PsycINFO Database Record

Endophytic fungi associated with Taxus fuana (West Himalayan Yew) of Pakistan: potential bio-resources for cancer chemopreventive agents.

CONTEXT: Endophytic fungi, being a prolific source of bioactive secondary metabolites, are of great interest for natural product discovery. OBJECTIVE: Isolation and partial characterization of endophytic fungi inhabiting the leaves and woody parts of Taxus fuana Nan Li & R.R. Mill. (Taxaceae) and evaluation of biological activity. MATERIALS AND METHODS: Endophytic fungal isolates were identified by molecular analysis of internal transcribed spacer (ITS) regions of 18S rDNA. Extracts of the endophytic fungi cultured on potato dextrose agar and modified medium were evaluated using cancer chemoprevention bioassays [inhibition of TNF-α-induced NFκB, aromatase and inducible nitric oxide synthase (iNOS); induction of quinone reductase 1 (QR1)] and growth inhibition with MCF-7 cells. RESULTS: Nine of 15 fungal isolates were identified as belonging to Epicoccum, Mucor, Penicillium, Chaetomium, Paraconiothriym, Plectania or Trichoderma. Five of the 15 extracts inhibited NFκB activity (IC50 values ranging between 0.18 and 17 µg/mL) and five inhibited iNOS (IC50 values ranging between 0.32 and 12.9 µg/mL). In the aromatase assay, only two isolates mediated inhibition (IC50 values 12.2 and 10.5 µg/mL). With QR1 induction, three extracts exhibited significant activity (concentrations to double activity values ranging between 0.20 and 5.5 µg/mL), and five extracts inhibited the growth of MCF-7 cells (IC50 values ranging from 0.56 to 17.5 µg/mL). Six active cultures were derived from woody parts of the plant material. CONCLUSION: The endophytic fungi studied are capable of producing pharmacologically active natural compounds. In particular, isolates derived from the wood of Taxus fuana should be prioritized for the isolation and characterization of bioactive constituents.