World federation of interventional and therapeutic neuroradiology (WFITN) federation assembly neurointerventional surgery safety checklist.

Over the last 10 years, there has been a rise in neurointerventional case complexity, device variety and physician distractions. Even among experienced physicians, this trend challenges our memory and concentration, making it more difficult to remember safety principles and their implications. Checklists are regarded by some as a redundant exercise that wastes time, or as an attack on physician autonomy. However, given the increasing case and disease complexity along with the number of distractions, it is even more important now to have a compelling reminder of safety principles that preserve habits that are susceptible to being overlooked because they seem mundane. Most hospitals have mandated a pre-procedure neurointerventional time-out checklist, but often it ends up being done in a cursory fashion for the primary purpose of 'checking off boxes'. There may be value in iterating the checklist to further emphasize safety and communication. The Federation Assembly of the World Federation of Interventional and Therapeutic Neuroradiology (WFITN) decided to construct a checklist for neurointerventional cases based on a review of the literature and insights from an expert panel.

Salinity-Induced Physiological Changes in Pea (Pisum sativum L.): Germination Rate, Biomass Accumulation, Relative Water Content, Seedling Vigor and Salt Tolerance Index.

Salinity affects and limits the yield potential of pulse crops. Therefore, an experiment was conducted to evaluate the salinity-induced physiological response of field peas by estimating the germination rate (%), accumulation of biomass, relative water content, and seedling vigor and salt tolerance index. The treatments included four salinity levels (NaCl) (i.e., 0 (control), 8, 12, and 16 dS m-1, respectively) and eight field pea genotypes (i.e., BD4175, BD4182, BD4225, BD6944, BD4176, BD4193, BD4493, and BD4496). All treatments were arranged in a factorial completely randomized design and repeated four times. Results indicated that the percentage and rate of germination, percentage reduction of fresh and dry weight, relative water content, seedling vigor index, and salt tolerant index of all genotypes of field peas were influenced significantly by the different levels of salinity. The radicle and plumule of all field pea genotypes were damaged by applying 12 and 16 dS m-1 salt stress. However, among these eight pea genotypes, two genotypes, namely BD4175 and BD4225, performed better under the 8 dS m-1 level of salinity and these two genotypes may be recommended for cultivation in field conditions of saline coastal areas of Bangladesh, and can also be used in future breeding programs for the development of salt-tolerant pea cultivars.

Genetic Characterisation of Colistin Resistant Klebsiella pneumoniae Clinical Isolates From North India.

Background: Increasing use of colistin has led to the world-wide emergence of mobile colistin resistant gene (mcr). The present study aimed to identify and characterise mcr and other drug-resistant genes in colistin resistant Klebsiella pneumoniae clinical isolates. Methods: Twenty-two colistin resistant K. pneumoniae were analysed for mcr and other drug-resistant genes, efflux pumps, and virulence genes, and for their biofilm forming ability. Pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) were performed for all mcr-1 positive isolates. S1-PFGE and Southern hybridisation were performed for localisation of mcr-1 and blaNDM. Results: Nineteen colistin resistant K. pneumoniae harboured mcr-1 and 3 had mgrB disruption. All isolates harboured blaOXA-48-type and ESBL genes; eight strains (five with mcr-1 and three with mgrB disruption) co-harboured blaNDM. Efflux pumps genes AcrAB and mdtK were detected in all 22 and tol-C in 21 isolates. Virulence-related genes entB and irp-1 were detected in all 22, mrkD in 20, and fimH-1 in 18 isolates; 11 isolates were strong biofilm producers. PFGE clustered mcr-1 positive isolates into eight groups based on ≥90% similarity; MLST revealed diverse sequence types, predominant being ST-15 (n = 4) and ST-16 (n = 4). Both mcr-1 and blaNDM were localised on plasmid and chromosome; mcr-1 was present on IncFII type and blaNDM on IncFIB and IncA/C type plasmids. Conclusions: Colistin resistance in K. pneumoniae was predominantly mediated by mcr-1. Co-existence of colistin, carbapenem, and other drug-resistant genes along with efflux pumps indicates towards enormous genomic plasticity in K. pneumoniae with ability to emerge as super-spreader of drug-resistance.

Novel variant NDM-11 and other NDM-1 variants in multidrug-resistant Escherichia coli from South India.

OBJECTIVES: Carbapenem resistance mediated by New Delhi metallo-ß-lactamase 1 (NDM-1) and its variants has caused a major public-health concern worldwide. Here we report for the first time an Escherichia coli isolate positive for a novel variant (NDM-11). METHODS: blaNDM genes were investigated in E. coli by PCR and sequencing, and blaNDM variants were further characterised. The susceptibility pattern of novel blaNDM-11 towards different antimicrobials was compared with blaNDM-1 by cloning and expression in E. coli TOP10. RESULTS: A total of 33 carbapenem-resistant E. coli isolates were screened by PCR for the presence of blaNDM, of which 15 (45.5%) were positive. Sequencing of the PCR products revealed 10 isolates with NDM-1 and 5 isolates with NDM variants (one each of NDM-4, NDM-8 and NDM-11 and two NDM-5). Other resistance genes, including blaTEM-1, blaCTX-M-15, blaVIM, plasmid-encoded AmpC blaCMY-2 and 16S methyltransferases (rmtB and rmtC), were also associated with NDM variants in different combinations. The blaNDM variants were located on a transferable IncF-type plasmid of >100kb. Pulsed-field gel electrophoresis (PFGE) showed that all five E. coli isolates were unrelated, and multilocus sequence typing (MLST) revealed that they all belonged to ST131. Expression of the blaNDM-1 and blaNDM-11 genes in E. coli TOP10 showed no significant difference in MICs to various ß-lactams, including carbapenems. CONCLUSIONS: This study underlines the spread of NDM variants with other antimicrobial resistance genes in E. coli in South India. It also describes a novel NDM variant (blaNDM-11) having an antimicrobial resistance pattern similar to blaNDM-1.

Prevalence and Molecular Characterization of New Delhi Metallo-Beta-Lactamases in Multidrug-Resistant Pseudomonas aeruginosa and Acinetobacter baumannii from India.

New Delhi metallo-beta-lactamase (NDM)-mediated carbapenem resistance in Pseudomonas aeruginosa and Acinetobacter baumannii is a major concern. We investigated the presence of NDM and its variants in P. aeruginosa and A. baumannii at a tertiary hospital in North India. A total of 236 isolates (130 P. aeruginosa and 106 A. baumannii) were included; 38 (29.23%) P. aeruginosa and 20 A. baumannii isolates (18.8%) were resistant to carbapenems and all of them were blaNDM positive. All 38 carbapenem-resistant P. aeruginosa harbored blaNDM-1, while 12 (60%) of 20 A. baumannii harbored blaNDM-2. Pulsed-field gel electrophoresis showed that all 58 isolates were clonally unrelated. By Southern blot analysis, blaNDM-2 was located on chromosome. The blaNDM-2-positive isolates were more frequently recovered from tracheal aspirate (67% vs.16%; p = 0.02) and intensive care unit (67% vs. 20%; p = 0.001) than blaNDM-1. Among other carbapenemases, VIM was significantly associated with blaNDM-1 than blaNDM-2 (61% vs. 17%; p = 0.006). Mortality between blaNDM-1- and blaNDM-2-infected patients was comparable. When expressed in Escherichia coli, blaNDM-2 transformant conferred one doubling dilution higher MIC value for cefotaxime, piperacillin/tazobactam than blaNDM-1. The study shows the emergence of blaNDM-mediated resistance among P. aeruginosa and A. baumannii and rapid evolution of blaNDM-2 in A. baumannii with its chromosomal localization.

Survivability and fitness cost of heterogeneous vancomycin-intermediate Staphylococcus aureus.

The aim of this study was to observe the survivability and fitness cost of heterogeneous vancomycin-intermediate Staphylococcus aureus(hVISA) isolates. Survivability study was performed on dry cotton swab, and fitness cost was evaluated by estimating growth kinetics and generation time constant in BACTEC automated system. Total mean maximum time of recovery on primary culture was 4.1 and 7.1 weeks (P = 0.0001) for hVISA and vancomycin-sensitive S. aureus (VSSA), respectively, in dry starved condition. No significant difference between the mean value of lag phase duration (P = 0.89) was noted between hVISA and VSSA isolate in growth kinetics. However, we observed lesser generation time of hVISA isolates compared to S. aureus ATCC 29213 (P = 0.0076). This study concluded that a significant difference in generation time between VSSA and hVISA and suggests that hVISA have fitness cost compared to VSSA. However, further studies with more cases are required.

High frequency of SCCmec type V and agr type I among heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) in north India.

OBJECTIVES: The objective of this study was to compare the genetic features of heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) and vancomycin-sensitive methicillin-resistant S. aureus (VS-MRSA) isolates. METHODS: The presence of staphylococcal cassette chromosome mec (SCCmec) types, Panton-Valentine leukocidin (PVL), accessory gene regulator (agr) types, and vanA and vanB genes in hVISA and VS-MRSA isolates was evaluated by PCR. Genetic relatedness was studied by pulsed-field gel electrophoresis (PFGE). RESULTS: The distribution of SCCmec types in hVISA was as follows: 13/29 (44.8%) each of types II and V, 1/29 (3.4%) type III and 2/29 (6.9%) type IVa. Among VS-MRSA isolates, 20/50 (40.0%) were SCCmec type II, 17/50 (34.0%) were type III, 3/50 (6.0%) were type IVa and 10/50 (20.0%) were type V. SCCmec type V was significantly associated with hVISA, whereas SCCmec type III showed an association with VS-MRSA (P=0.020 and P=0.001, respectively). The PVL gene was detected in 9/29 hVISA (31.0%) and 13/50 VS-MRSA (26.0%). By PFGE analyses, both hVISA and VS-MRSA strains were found to be clonally unrelated. In hVISA isolates, 24/29 (82.8%) were agr type I, 3/29 (10.3%) were type III and 2/29 (6.9%) were non-typeable. However, in VS-MRSA isolates, 25/50 (50.0%) were type II, 15/50 (30.0%) were type I, 7/50 (14.0%) were type III and 3/50 (6.0%) were non-typeable. CONCLUSIONS: The study shows that healthcare-associated MRSA strains may harbour community-acquired MRSA genetic markers. The changing molecular epidemiology and role of agr I in reduced vancomycin susceptibility in hVISA requires further investigation.

Increasing Trend of Heterogeneous Vancomycin Intermediate Staphylococcus aureus in a Tertiary Care Center of Northern India.

Heterogeneous vancomycin intermediate Staphylococcus aureus (hVISA) strains are increasingly reported, and their association with vancomycin treatment failure is a well-known problem worldwide. A total of 500 clinical isolates of methicillin-resistant S. aureus were screened for hVISA by four different methods from May 2011 to May 2014. The number of strains screened for hVISA from May to April in 2011-2012, 2012-2013, and 2013-2014 were 198, 123, and 179, respectively. hVISA strains were studied by transmission electron microscopy (TEM) for cell wall thickness and also for their ability to form biofilm on a polystyrene microtiter plate. hVISA strains detected by four different methods-brain heart infusion agar with vancomycin with 4 mg/L/gradient plate/macro E-test/and glycopeptide resistance detection (GRD) E test-were as follows: 11.6%/10%/9%, and 9.5% in 2011-2012, 12.1%/9.7%/8.9%, and 10.5% in 2012-2013, and 13.9%/11.7%/11.1%, and 12.8% in 2013-2014, respectively. Population analysis profile-area under curve analysis confirmed hVISA in 4.5% (9/198), 6.5% (8/123), and 6.7% (12/179) in respective years; 24% (7/29) of hVISA isolates were nonsusceptible to daptomycin. TEM showed a significant increase in cell wall thickness of hVISA isolates (p<0.001) with a distinct reduction in their biofilm formation ability.

RmtC and RmtF 16S rRNA Methyltransferase in NDM-1-Producing Pseudomonas aeruginosa.

We investigated 16S rRNA methyltransferases in 38 blaNDM-1-positive Pseudomonas aeruginosa isolates and found RmtC in 3 isolates, 1 of which also harbored RmtF. The isolates were clonally unrelated; rmtC and rmtF genes were located on a chromosome with the blaNDM-1 gene. Strategies are needed to limit the spread of such isolates.

Glycopeptide and daptomycin susceptibility trends among clinical isolates of methicillin-resistant Staphylococcus aureus in a tertiary care center in North India.

Increased vancomycin minimum inhibitory concentration (MIC) levels in Staphylococcus aureus and their association with vancomycin treatment failure are well-known problems. Few studies have recognized progressive increases in glycopeptide MIC levels for S. aureus strains in recent years. This study determined glycopeptide and daptomycin susceptibility among methicillin resistant S. aureus (MRSA) strains. A total of 776 clinical isolates of MRSA recovered from 2009 to 2012 were studied for glycopeptide and daptomycin susceptibility using the E-test method. The vancomycin MIC geometric mean (GM) of the MRSA isolates was 0.923, 0.944, 1.134 and 1.294 mg/L in the years 2009, 2010, 2011 and 2012, respectively, and the trend significantly increased over the years (P < 0.0001). Similarly, the teicoplanin MIC GM was 1.47, 1.49, 1.8 and 2.04 mg/L in the years from 2009 to 2012, respectively (P < 0.0001). MIC shifts were not found for daptomycin (P > 0.232). A significant increase in the MIC for glycopeptides was observed among the clinical MRSA isolates at our center over a 4-year period. However, the daptomycin MIC did not increase in the observed MRSA isolates.

Prevalence and molecular characterisation of New Delhi metallo-ß-lactamases NDM-1, NDM-5, NDM-6 and NDM-7 in multidrug-resistant Enterobacteriaceae from India.

The growing prevalence of carbapenem resistance in Enterobacteriaceae worldwide is a major concern. New Delhi metallo-ß-lactamase (NDM)-mediated carbapenem resistance has been identified in Enterobacteriaceae from numerous countries including those of the Indian subcontinent. Currently, seven NDM ß-lactamase variants (NDM-1 to -7) have been identified. This study evaluated the detection and molecular characterisation of NDM variants in Enterobacteriaceae at a tertiary care hospital in India. A total of 464 isolates were tested; 57 (12.3%) were resistant or showed reduced susceptibility to imipenem and meropenem. All carbapenem-resistant isolates were blaNDM-positive by PCR, but 13 isolates bore variants that differed in sequence from blaNDM-1. NDM-5, NDM-6 and NDM-7 were identified in two, eight and three isolates, respectively. blaNDM variants were located on plasmids of >100kb with IncF, IncA/C and untypeable replicon types. Genes encoding the 16S rRNA methyltransferases RmtB, RmtC and ArmA as well as those for AmpC ß-lactamases were also located on the same plasmids as blaNDM in different combinations. The prevalence of NDM-5 to -7 variants was significantly higher in Escherichia coli (P=0.015) and they were more frequently isolated from the urology ward (P=0.037) than NDM-1. The mortality rate was comparable between patients infected with isolates positive for blaNDM-1 and blaNDM variants [25% (11/44) vs. 23% (3/13)]. Expression of blaNDM variants in E. coli using the same promoter showed that NDM-7 conferred higher resistance to imipenem. The diverse genotypic features of blaNDM indicate rapid evolution of NDM resulting from their wide spread in the Indian subcontinent.

Evaluation of ELISA, neck muscle, tongue and eyelid examinations for the diagnosis of swine cysticercosis in a highly endemic area of north India.

Swine cysticercosis is very common in the developing countries where pigs are raised. Undercooked measly pork consumption leads to taeniasis; Taenia carriers act as source of human and swine cysticercosis and neurocysticercosis. Diagnosis of swine cysticercosis is important to break the cycle of disease transmission. The present study compared the neck muscle, tongue and eye examinations, and serum ELISA with different preparations (crude lysate, cyst fluid, scolex and cyst wall antigens) of Taenia solium cyst for the diagnosis of swine cysticercosis. Total of 24 pigs initially identified by neck muscle, tongue and eyelid examinations were purchased from local slaughter house and subjected to MRI for confirmation of cysticercosis. Sera from 20 MRI confirmed infected pigs and 50 disease free controls were subjected to ELISA with T. solium cyst antigens. Neck muscle examination was 100% sensitive and 75% specific for the diagnosis of swine cysticercosis, whereas tongue and eye examinations were 70% and 25% sensitive, respectively. ELISA with crude lysate had 85% sensitivity and 98% specificity. ELISA with cyst fluid, scolex and cyst wall antigens showed 70%, 65%, and 45% sensitivity, respectively. The present study showed that neck muscle examination was highly sensitive but less specific, while ELISA with crude antigens had reasonable sensitivity and high specificity for diagnosis of swine cysticercosis. ELISA with crude lysate can be used as a screening tool for swine infection.