Knowledge, practice, counseling confidence, and intention to use AAR model of smoking cessation among respiratory therapists: A cross-sectional study.

There is a paucity of research on knowledge, practice, counseling confidence, and intention to use ask, advice, and refer (AAR) model of smoking cessation among respiratory therapists (RTs). Thus, we aimed to analyze the characteristics and factors that may influence them. We collected data using online questionnaires from convenience sample of active licensed RTs in Saudi Arabia. We included 206 participants. A descriptive analysis of the demographic information and characteristics of smoking cessation counseling practices and confidence were conducted. Multiple linear regression was used to test whether demographic variables and AAR model components significantly predicted the RTs' calculated cumulative score of tobacco counseling confidence skills. Our results showed a deficiency in tobacco knowledge among RTs. Most RTs did not have certifications or attend lectures or seminars related to tobacco treatment. RTs were unfamiliar with the smoking cessation program contact information and mobile smoking cessation clinics but reported a high tobacco counseling confidence score. Clinical experience (P = .008), familiarity with smoking cessation program contact information (P = .02), inquiry regarding smoking status (P < .001), and advice regarding smoking status (P = .03) significantly predicted tobacco counseling confidence levels in RTs. RT experience, knowledge, and awareness of smoking cessation programs could enhance the confidence level among them in implementing AAR model.

On the formation chemistry of brominated polycyclic aromatic hydrocarbons (BrPAHs).

Brominated polycyclic aromatic hydrocarbons (BrPAHs) have been consistently detected in various environmental matrices, and measured at alarming rates in stack emissions. However, formation mechanisms and bromination patterns of BrPAHs remain unclear. This contribution constructs detailed mechanistic pathways for the synthesis of selected BrPAHs (namely bromine-bearing naphthalene, acenaphthylene, anthracene, and phenanthrene). Mapped-out pathways follow the Bittner-Howard's route in the hydrogen abstraction acetylene addition (HACA) mechanism, in which a second C2HBr molecule is added to the first one. Constructed kinetic model portrays temperature-dependent profiles of major and minor species. Direct loss of an H atom from the acetylenic fragment appears to be more important at elevated temperatures, when compared with further addition of C2HBr cuts or ring-cyclization reactions. The occurrence of closed-shell Diels-Alder pathway should be inhibited owing to sizable enthalpic barriers. Fukui Indices for electrophilic substitutions (f-1) establish bromination' s pattern of selected BrPAHs. The diradical character of BrPAHs coupled with electron-deficient C(Br) sites, render BrPAHs as potent precursors for the formation of environmentally persistent free radicals (EPFRs). Findings reported herein shall be useful in comprehending the formation chemistry of BrPAHs, a less-investigated category of toxicants in thermal systems.

Clostridium difficile infections in Finland, 2008-2015: trends, diagnostics and ribotypes.

We evaluated Clostridium difficile (CD) diagnostics in Finnish clinical microbiology laboratories during 2006-2011, with an update in 2015, in relation to CD surveillance data of the National Infectious Disease Register (NIDR) and ribotyping data from the national reference laboratory during the years 2008-2015. In 2011, diagnostic activity varied regionally more than three-fold and the positivity rate ranged between 7 and 21%. Nucleic acid amplification testing (NAAT) was implemented in the regions with high activity and NAAT users tested 30% more patients and found 15% more cases per population than those not using it. Culture was performed in 79% of laboratories, primary toxin testing by enzyme immunoassay (EIA) in 83% and by NAAT in 17%. In 2014, 12/19 laboratories used NAAT as the primary detection method and four as the secondary method, and ten cultured. Increasing usage of NAAT was not systematically related to various trends detected regionally in annual CD rates. Polymerase chain reaction (PCR) ribotyping of 1771 CD isolates (4.1% of CD cases) identified 146 distinct profiles, of which 37% were binary toxin positive. The most common ribotype was 027, but its proportion decreased, while 078 slightly increased. Transition from culture to NAAT in CD infection (CDI) diagnostics did not cause a significant increase in the observed CDI incidence. Major differences between diagnostic activity, methods and strategies in different regions have persisted over the years, which should be considered when comparing the regional epidemiology of CDI.

Incidence, case fatality and genotypes causing Clostridium difficile infections, Finland, 2008.

Since 2000, the epidemiology of C. difficile infections (CDI) has changed in the US and Europe. Few population-based assessments of both incidence and case fatality of CDI have been performed. In this study, the Finnish nationwide laboratory-based surveillance data from the year 2008 were analysed to assess the incidence and case fatality of CDI, and to detect regional differences in relation to molecular epidemiology. A total of 6201 episodes of CDI were identified (118.3/100 000 population; range by regions, 57.2-189.1). The incidence increased by age and was highest in persons aged >84 years (1286.0). Of the CDI episodes, 711 (11.5%; range by regions, 2.2-15.0%) led to death within 30 days. The 30-day case fatality was highest (22.0%) in persons aged >84 years. In total, 334 (5% of all episodes) isolates from 13/21 regions were sent for genotyping: 120 (36%) were of PCR ribotype 027, and it was found in 6/13 regions. Among the rest of the isolates, 53 (16%) were of type 001, and 19 (6%) of 002 and 014. The incidence and case fatality were highest in elderly persons and varied regionally. This may be explained by uneven spread of hypervirulent PCR ribotypes, such as 027, but also differences in diagnostic activity or the patient populations among which the outbreaks are occurring.

Comparison of repetitive extragenic palindromic sequence-based PCR with PCR ribotyping and pulsed-field gel electrophoresis in studying the clonality of Clostridium difficile.

Clostridium difficile infection is most often induced by antibiotic treatment. Recently, morbidity and mortality resulting especially from C. difficile PCR ribotype 027 have increased significantly. In addition, more severe disease has been associated with C. difficile PCR ribotype 078 strains. Thus, reliable typing methods for epidemic control are needed. In the present study, we compared an automated repetitive extragenic palindromic sequence-based PCR (rep-PCR) method (DiversiLab; Bacterial Barcodes, Inc., Athens, GA, USA) to PCR ribotyping and pulsed-field gel electrophoresis (PFGE) typing using 205 isolates of C. difficile (including 24 previously characterized isolates). Among the 181 clinical isolates, a total of 31 different PCR ribotypes, 38 different PFGE types and subtypes and 28 different rep-PCR types were found. Six major rep-PCR groups (DL1-DL6) harboured 86% of the clinical isolates. All isolates belonging to PCR ribotypes 027 and 001 clustered in their own rep-PCR groups, enabling us to screen out the hypervirulent ribotype 027 strain. Within the PCR ribotype 001, four subgroups were found using rep-PCR. Overall, in 75% (135/181) of the isolates, the classification attributed following rep-PCR and PCR ribotyping was comparable. In conclusion, the automated rep-PCR-based typing method represents an option for first-line molecular typing in local clinical microbiology laboratories. The method was easy to use as well as rapid, requiring less hands-on time than PCR ribotyping or PFGE typing. The conventional PCR ribotyping or PFGE, however, are needed for confirmatory molecular epidemiology. In addition, more epidemiology-oriented studies are needed to examine the discriminatory power of automated rep-PCR with isolates collected from a larger geographical area and during a longer period of time.

Concomitant mutations and splice variants in KRAS and BRAF demonstrate complex perturbation of the Ras/Raf signalling pathway in advanced colorectal cancer.

BACKGROUND AND AIMS: KRAS and BRAF mutations occur in colorectal cancers (CRCs) and are considered mutually exclusive methods of activating the RAS/RAF/MEK/ERK pathway. This pathway is a therapeutic target and KRAS mutation may predict tumour responsiveness. The purpose of this study was to investigate the relationship between KRAS and BRAF mutations in 24 CRC cell lines and 29 advanced CRCs. METHODS: KRAS and BRAF mutations were detected using high resolution melting and sequencing. Expression of mutations was confirmed by reverse transcription- PCR (RT-PCR) and sequencing. CpG island methylator phenotype (CIMP) was tested by methylation-specific PCR. RESULTS: KRAS or BRAF mutation occurred in 79% of cell lines and 59% of CRCs. In the cell lines, KRAS mutations occurred in 54% of cases (with 62% in codons 12/13 and 38% in other codons). Four cell lines had a homozygous mutation. Only heterozygous BRAF mutations were detected in 29% cell lines. The V600E mutation occurred most commonly and was associated with CIMP+ status (p = 0.005). Mutations at codons 529 and 581 were also found and, in one case, BRAF and KRAS mutation co-occurred. Unexpectedly, BRAF splice variants (with a predicted kinase-dead protein) were found in 5/24 (21%) cell lines. In advanced CRCs, KRAS mutations occurred in 48% of cases (64% codons 12/13, 36% other codons) and BRAF mutations in 10% (66% V600E, 33% exon 11). A compound KRAS/BRAF mutation was not seen. CONCLUSIONS: Disrupted Ras/Raf signalling is common in CRC. Homozygous KRAS mutations and concomitant KRAS/BRAF mutations may be indicative of a gene dosage effect. The significance of BRAF splice variants is uncertain but may represent another layer of complexity. Finally, if KRAS mutation is to be used for predictive testing, then the whole gene may need to be screened as mutations occur outside codons 12/13.

Molecular characterization of methicillin-resistant Staphylococcus epidermidis strains from bacteraemic patients.

In order to study the clonality of clinical methicillin-resistant Staphylococcus epidermidis (MRSE) strains and their staphylococcal cassette chromosome mec (SCCmec) elements, 60 isolates of MRSE from bacteraemic patients in three units of the Helsinki University Hospital, Finland were selected, covering the periods 1990-1993 and 1997-1998. The MRSE strains were analysed by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing and SCCmec typing. Eleven PFGE types (FIN-SE-1-11) with sequence type ST2 (clonal complex 2; CC2) were identified. The previously established methicillin-resistant Staphylococcus aureus SCCmec criteria were applied to name the MRSE SCCmec complexes, and it was found that 7% of the isolates carried SCCmec type IA (ccrA1, class B), whereas the majority (93%) yielded six non-typeable SCCmec PCR patterns (P1-P6). Within each SCCmec PCR pattern, two ccr recombinase genes (ccrA2 and ccrA3) and two mec gene complexes (class A and class B) were detected. In addition, the ccrC gene was associated with three of the six patterns. In conclusion, the MRSE strains were genetically related to each other (ST2) but their SCCmec complexes were unique combinations of elements previously recognized among SCCmec types III and IV.

Clonality of epidemic methicillin-resistant Staphylococcus aureus strains in Finland as defined by several molecular methods.

In Finland, the incidence of methicillin-resistant Staphylococcus aureus (MRSA) strains has increased ten fold within the last decade. In order to follow the changing epidemiology of MRSA, accurate typing of S. aureus strains is important. The purpose of this study was to reanalyse 44 previously recognised Finnish epidemic MRSA strains (EMRSA) by several molecular typing methods and to revise their nomenclature. The 44 EMRSA strains were grouped into 26 pulsed-field gel electrophoresis (PFGE) clusters, 20 multi locus sequence typing (MLST) sequence types (ST) belonging to 12 clonal complexes (CC) of which CC8 was the most prevalent, and 27 spa types belonging to four clonal complexes. The staphylococcal cassette chromosome mec (SCCmec) type IV was predominant, and 48% of the strains were nonmultiresistant to antibiotics. The discriminatory power of PFGE clusters, MLST, and spa typing was high. The overall concordance values of typing methods differed when assessed by two different methods. Adjusted Rand coefficient provided fairly low correlations for all comparisons. However, spa type was able to efficiently predict types and clonal complexes of most of the other methods with high probability (> or =80%).

Panton-Valentine leukocidin genes and staphylococcal chromosomal cassette mec types amongst Finnish community-acquired methicillin-resistant Staphylococcus aureus strains, 1997-1999.

Methicillin-resistant Staphylococcus aureus (MRSA) strains from Finland covering years 1997-1999 were studied for the presence of Panton-Valentine leukocidin (PVL) gene loci, and the clinically well-defined community-acquired MRSA (CA-MRSA) strains (n = 108) also for staphylococcal chromosomal cassette mec (SCCmec) and multilocus sequence types (MLST). Only a minority (12%) of the CA-MRSA strains contained the PVL gene loci and possessed genotypes formerly described as typical to CA-MRSA strains. The majority of these strains were heterogenous by MLST and pulsed-field gel electrophoresis (PFGE) analysis but, however, harboured the SCCmec cassette type IV. In conclusion, it seems doubtful to consider only molecular characteristics such as the presence of PVL genes as definite markers for CA-MRSA strains.

Diagnosis of Sex Chromosome Disorders and Prenatal Diagnosis of Down Syndrome using Interphase Fluorescent In-Situ Hyperidization Technique.

BACKGROUND: Thousands of infants are born each year with chromosomal abnormalities that severely impact physical and mental development. Among common genetic disorders are Down syndrome (trisomy 21) and sex chromosomal disorders. OBJECTIVES: Evaluation of guidelines used for prenatal diagnosis of Down syndrome (DS) as well as sex chromosomal disorders including interphase Fluorescent In Situ Hyperidization (FISH) technique. METHODS: Enrolled cases were among those presenting to Genetics and Neonatology Units, Mansoura and Ain-Shams University hospitals,(Egypt) during 2002 to 2004. These included: Groups 1 comprised fifty pregnant women presenting for genetic counseling. They were subjected to complete history analysis, ultrasound examination in addition to triple screening test (for alpha feto protein (AFP), human chorionic goandotrophin (HCG) and unconjugated esteriol (E2). Results were confirmed by doing routine karyogram on cultured amniotic fluid. Groups 2 comprised suspected cases with sex chromosomal disorders including neonates with ambiguous genitalia (64 cases) and adults with primary amenorrhea (69 cases) or infertility (38 cases). They were subjected to a diagnostic workup including RESULTS: Among the pregnant women group, seven were found to be at a high risk of having DS fetuses including 3 cases with a history of affected off-springs, 2 cases with age above 35 years, and 2 cases with high triple test. Only one case had positive trisomy 21 on interphase FISH confirmed by karyogram on cultured amniotic cells. The other 6 ladies had normal FISH confirmed by karyograms. Regarding the other group, 5 cases out of the 9 females were proved to be feminized males, one proved mosaic turner, one proved mixed gonadal dysgenesis and 2 normal females. On the other hand one out of three males were proved to be verilized female while the other one was a male with incomplete testicular feminization and the last one was a male with infertility diagnosed as Klinefelter syndrome at the age of 26 years. CONCLUSION: Interphase FISH is a rapid, accurate and very sensitive method in sex chromosom and autosomal abnormalities. It adds to the diagnostic utility of routine cytogenetics and its use on interphase nuclei overcomes the difficulty of conventional cytogenetics. It could be used in the prenatal diagnosis of DS in addition to ultrasonography, and triple test.

Comparison of genotypes of methicillin-resistant and methicillin-sensitive Staphylococcus aureus in Finland.

The frequency of horizontal transfer of the staphylococcal cassette chromosome mec to methicillin-susceptible Staphylococcus aureus is unknown. In order to gain more information regarding this frequency in Finland, the genotypes of 299 clinical methicillin-sensitive Staphylococcus aureus isolates were compared to representatives of 24 epidemic methicillin-resistant Staphylococcus aureus genotypes. Sixty-eight percent of the methicillin-sensitive isolates had a genotype similar to eight of the epidemic methicillin-resistant strains. The remaining isolates (32%) showed 22 different genotypes. The results indicate that, in Finland, several methicillin-sensitive Staphylococcus aureus genotypes may have acquired the staphylococcal cassette chromosome mec.