bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification, characterization, and chromosomal organization of cell division cycle genes in Caulobacter crescentus
Authors: Ohta N , Ninfa A J , Allaire A , Kulick L , Newton A
Published: November 2016

We report a detailed characterization of cell division cycle (cdc) genes in the differentiating gram-negative bacterium Caulobacter crescentus. A large set of temperature-sensitive cdc mutations was isolated after treatment with the chemical mutagen N-methyl-N'-nitro-N-nitrosoguanidine. Analysis of independently isolated mutants at the nonpermissive temperature identified a variety of well-defined terminal phenotypes, including long filamentous cells blocked at various stages of the cell division cycle and two unusual classes of mutants with defects in both cell growth and division. The latter strains are uniformly arrested as either short bagel-shaped coils or large predivisional cells. The polar morphology of these cdc mutants supports the hypothesis that normal cell cycle progression is directly responsible for developmental regulation in C. crescentus. Genetic and physical mapping of the conditional cdc mutations and the previously characterized dna and div mutations identified at least 21 genes that are required for normal cell cycle progression. Although most of these genes are widely scattered, the genetically linked divA, divB, and divE genes were shown by genetic complementation and physical mapping to be organized in one gene cluster at 3200 units on the chromosome. DNA sequence analysis and marker rescue experiments demonstrated that divE is the C. crescentus ftsA homolog and that the ftsZ gene maps immediately adjacent to ftsA. On the basis of these results, we suggest that the C. crescentus divA-divB-divE(ftsA)-ftsZ gene cluster corresponds to the 2-min fts gene cluster of Escherichia coli.
Restrictive glycosylphosphatidylinositol anchor synthesis in cwh6/gpi3 yeast cells causes aberrant biogenesis of cell wall proteins
Authors: Vossen J H , Müller W H , Lipke P N , Klis F M
Published: November 2016

We previously reported that the defects in the Saccharomyces cerevisiae cwh6 Calcofluor white-hypersensitive cell wall mutant are caused by a mutation in SPT14/GPI3, a gene involved in glycosylphosphatidylinositol (GPI) anchor biosynthesis. Here we describe the effect of cwh6/spt14/gpi3 on the biogenesis of cell wall proteins. It was found that the release of precursors of cell wall proteins from the endoplasmic reticulum (ER) was retarded. This was accompanied by proliferation of ER structures. The majority of the cell wall protein precursors that eventually left the ER were not covalently incorporated into the cell wall but were secreted into the growth medium. Despite the inefficient incorporation of cell wall proteins, there was no net effect on the protein level in the cell wall. It is postulated that the availability of GPI-dependent cell wall proteins determines the rate of cell wall construction and limits growth rate.
Sequence analysis and characterization of a 40-kilodalton Borrelia hermsii glycerophosphodiester phosphodiesterase homolog
Authors: Shang E S , Skare J T , Erdjument-Bromage H , Blanco D R , Tempst P , Miller J N , Lovett M A
Published: November 2016

We report the purification, molecular cloning, and characterization of a 40-kDa glycerophosphodiester phosphodiesterase homolog from Borrelia hermsii. The 40-kDa protein was solubilized from whole organisms with 0.1% Triton X-100, phase partitioned into the Triton X-114 detergent phase, and purified by fast-performance liquid chromatography (FPLC). The gene encoding the 40-kDa protein was cloned from a B. hermsii chromosomal DNA lambda EXlox expression library and identified by using affinity antibodies generated against the purified native protein. The deduced amino acid sequence included a 20-amino-acid signal peptide encoding a putative leader peptidase II cleavage site, indicating that the 40-kDa protein was a lipoprotein. Based on significant homology (31 to 52% identity) of the 40-kDa protein to glycerophosphodiester phosphodiesterases of Escherichia coli (GlpQ), Bacillus subtilis (GlpQ), and Haemophilus influenzae (Hpd; protein D), we have designated this B. hermsii 40-kDa lipoprotein a glycerophosphodiester phosphodiesterase (Gpd) homolog, the first B. hermsii lipoprotein to have a putative functional assignment. A nonlipidated form of the Gpd homolog was overproduced as a fusion protein in E. coli BL21(DE3)(pLysE) and was used to immunize rabbits to generate specific antiserum. Immunoblot analysis with anti-Gpd serum recognized recombinant H. influenzae protein D, and conversely, antiserum to H. influenzae protein D recognized recombinant B. hermsii Gpd (rGpd), indicating antigenic conservation between these proteins. Antiserum to rGpd also identified native Gpd as a constituent of purified outer membrane vesicles prepared from B. hermsii. Screening of other pathogenic spirochetes with anti-rGpd serum revealed the presence of antigenically related proteins in Borrelia burgdorferi, Treponema pallidum, and Leptospira kirschneri. Further sequence analysis both upstream and downstream of the Gpd homolog showed additional homologs of glycerol metabolism, including a glycerol-3-phosphate transporter (GlpT), a glycerol-3-phosphate dehydrogenase (GlpD), and a thioredoxin reductase (TrxB).
aarC, an essential gene involved in density-dependent regulation of the 2'-N-acetyltransferase in Providencia stuartii
Authors: Rather P N , Solinsky K A , Paradise M R , Parojcic M M
Published: November 2016

The 2'-N-acetyltransferase [AAC(2')-Ia] in Providencia stuartii has a dual function where it is involved in the acetylation of peptidoglycan and certain aminoglycosides. A search for negative regulators of the aac(2')-Ia gene has resulted in the identification of aarC. A missense allele (aarC1) resulted in an 8.9-fold increase in beta-galactosidase accumulation from an aac(2')-lacZ transcriptional fusion. Northern blot analysis demonstrated an increase in aac(2')-Ia mRNA accumulation that was specific to cells at high density. In addition, the aarC1 allele also resulted in a substantial increase in the expression of aarP, a transcriptional activator of the aac(2')-Ia gene. The wild-type aarC gene was isolated by complementation and encodes a predicted protein of 365 amino acids with a molecular mass of 39,815 Da. The predicted AarC protein exhibited 88% amino acid homology to the previously identified GcpE protein of Escherichia coli and 86% homology to a gene product from Haemophilus influenzae. The E. coli gcpE gene was able to functionally complement the aarC1 allele in P. stuartii. The aarC1 allele was identified as a T to G transversion that resulted in a valine to glycine substitution at position 136 in the AarC protein. The aarC gene appears to be essential for cell viability as construction of a disrupted copy (aarC::lacZ) was possible only in cells that carried an episomal copy of aarC or gcpE.
Protein tyrosine phosphorylation in the cyanobacterium Anabaena sp. strain PCC 7120
Authors: McCartney B , Howell L D , Kennelly P J , Potts M
Published: November 2016

Components of a protein tyrosine phosphorylation/dephosphorylation network were identified in the cyanobacterium Anabaena sp. strain PCC 7120. Three phosphotyrosine (P-Tyr) proteins of 27, 36, and 52 kDa were identified through their conspicuous immunoreactions with RC20H monoclonal antibodies specific for P-Tyr. These immunoreactions were outcompeted completely by free P-Tyr (5 mM) but not by phosphoserine or phosphothreonine. The P-Tyr content of the three major P-Tyr proteins and several minor proteins increased with their time of incubation in the presence of Mg-ATP and the protein phosphatase inhibitors sodium orthovanadate and sodium fluoride. Incubation of the same extracts with [gamma-32P]ATP but not [alpha-32P]ATP led to the phosphorylation of five polypeptides with molecular masses of 20, 27, 52, 85, and 100 kDa. Human placental protein tyrosine phosphatase 1B, with absolute specificity for P-Tyr, liberated significant quantities of 32Pi from four of the polypeptides, confirming that a portion of the protein-bound phosphate was present as 32P-Tyr. Alkaline phosphatase and the dual-specificity protein phosphatase IphP from the cyanobacterium Nostoc commune UTEX 584 also dephosphorylated these proteins and did so with greater apparent efficiency. Two of the polypeptides were partially purified, and phosphoamino analysis identified 32P-Tyr, [32P]phosphoserine, and [32P]phosphothreonine. Anabaena sp. strain PCC 7120 cell extracts contained a protein tyrosine phosphatase activity that was abolished in the presence of sodium orthovanadate and inhibited significantly by the sulfhydryl-modifying agents p-hydroxymercuriphenylsulfonic acid and p-hydroxymercuribenzoate as well as by heparin. In Anabaena sp. strain PCC 7120 the presence and/or phosphorylation status of P-Tyr proteins was influenced by incident photon flux density.
Characterization of a temperature-sensitive Escherichia coli mutant and revertants with altered seryl-tRNA synthetase activity
Authors: Ferri M L , Vincent C , Leberman R , Härtlein M
Published: November 2016

A mutation in the structural gene coding for seryl-tRNA synthetase in temperature-sensitive Escherichia coli K28 has been reported to alter the level of enzyme expression at high temperature (R. J. Hill and W. Konigsberg, J. Bacteriol. 141:1163-1169, 1980). We identified this mutation as a C-->T transition in the first base of codon 386, resulting in a replacement of histidine by tyrosine. The steady-state levels of serS mRNA in K28 and in the wild-type strains are very similar. Pulse-chase labeling experiments show a difference in protein stability, but not one important enough to account for the temperature sensitivity of K28. The main reason for the temperature sensitivity of K28 appears to be the low level of specific activity of the mutant synthetase at nonpermissive temperature, not a decreased expression level. Spontaneous temperature-resistant revertants were selected which were found to have about a fivefold-higher level of SerRS than the K28 strain. We identified the mutation responsible for the reversion as being upstream from the -10 sequence in the promoter region. The steady-state levels of serS mRNA in the revertants are significantly higher than that in the parental strain.
The Agrobacterium tumefaciens virulence gene chvE is part of a putative ABC-type sugar transport operon
Authors: Kemner J M , Liang X , Nester E W
Published: November 2016

The Agrobacterium tumefaciens virulence determinant ChvE is a periplasmic binding protein which participates in chemotaxis and virulence gene induction in response to monosaccharides which occur in the plant wound environment. The region downstream of the A. tumefaciens chvE gene was cloned and sequenced for nucleotide and expression analysis. Three open reading frames transcribed in the same direction as chvE were revealed. The first two, together with chvE, encode putative proteins of a periplasmic binding protein-dependent sugar uptake system, or ABC-type (ATP binding cassette) transporter. The third open reading frame encodes a protein of unknown function. The deduced transporter gene products are related on the amino acid level to bacterial sugar transporters and probably function in glucose and galactose uptake. We have named these genes gguA, -B, and -C, for glucose galactose uptake. Mutations in gguA, gguB, or gguC do not affect virulence of A. tumefaciens on Kalanchoe diagremontiana; growth on 1 mM galactose, glucose, xylose, ribose, arabinose, fucose, or sucrose; or chemotaxis toward glucose, galactose, xylose, or arabinose.
High-level genetic diversity in the vapD chromosomal region of Helicobacter pylori
Authors: Cao P , Cover T L
Published: November 2016

Helicobacter pylori isolates from different patients are characterized by diversity in the nucleotide sequences of individual genes, variation in genome size, and variation in gene order. Genetic diversity is particularly striking in vacuolating cytotoxin (vacA) alleles. In this study, five open reading frames (ORFs) were identified within a 4.2-kb region downstream from vacA in H. pylori 60190. One of these ORFs was closely related to the virulence-associated protein D (vapD) gene of Dichelobacter nodosus (64.9% nucleotide identity). A probe derived from vapD of H. pylori 60190 hybridized with only 19 (61.3%) of 31 H. pylori strains tested. Sequence analysis of the vapD region in vapD-negative H. pylori strains revealed that there were two different families of approximately 0.5-kb DNA segments, which were both unrelated to vapD. The presence of vapD was not associated with any specific family of vacA alleles. These findings are consistent with a recombinational population structure for H. pylori.
Identification and characterization of Myxococcus xanthus mutants deficient in calcofluor white binding
Authors: Ramaswamy S , Dworkin M , Downard J
Published: November 2016

Calcofluor white is a fluorescent dye that binds to glycans and can be used to detect extracellular polysaccharide in Myxococcus xanthus and many other bacteria. We observed that an esg mutant showed less binding to calcofluor white than wild-type cells. Unlike S-motility mutants that share this phenotypic characteristic, the esg mutant exhibited S motility. This led us to identify a collection of nine new transposon insertion mutants, designated Cds (for calcofluor white binding deficient and S motile), which exhibited a phenotype similar to that of the esg strain. The Cds phenotype was found in 0.6% of the random insertion mutants that were screened. The Cds mutants were also found to be defective in cell-cell agglutination and developmental aggregation. Extracellular matrix fibrils composed of roughly equal amounts of polysaccharide and protein have been shown to be involved in agglutination, and electron microscopic examination showed that esg and the other Cds mutants lack the wild-type level of fibrils. Analysis of total M. xanthus carbohydrate demonstrated that polysaccharide content increased by about 50% when wild-type cells entered stationary phase. This induction was reduced or eliminated in all of the Cds mutants. The degree of polysaccharide deficiency in the Cds mutants correlated with the degree of loss of agglutination and dye binding as well as with the severity of the developmental aggregation defect. Preliminary genetic characterization demonstrated that the transposon insertion mutations in three of the Cds mutants (SR53, SR171, and SR200) were loosely linked. The results of this study suggest that many genes are involved in the production of calcofluor white binding polysaccharide material found in the extracellular matrix and that the polysaccharide is fibrillar. These results are also consistent with the findings of earlier studies which indicated that fibrils function to join agglutinating cells and to form multicellular fruiting aggregates.
Isolation of mutants of Acinetobacter calcoaceticus deficient in wax ester synthesis and complementation of one mutation with a gene encoding a fatty acyl coenzyme A reductase
Authors: Reiser S , Somerville C
Published: November 2016

Acinetobacter calcoaceticus BD413 accumulates wax esters and triacylglycerol under conditions of mineral nutrient limitation. Nitrosoguanidine-induced mutants of strain BD413 were isolated that failed to accumulate wax esters under nitrogen-limited growth conditions. One of the mutants, Wow15 (without wax), accumulated wax when grown in the presence of cis-11-hexadecenal and hexadecanol but not hexadecane or hexadecanoic acid. This suggested that the mutation may have inactivated a gene encoding either an acyl-acyl carrier protein or acyl-coenzyme A (CoA) reductase. The Wow15 mutant was complemented with a cosmid genomic library prepared from wild-type A. calcoaceticus BD413. The complementary region was localized to a single gene (acr1) encoding a protein of 32,468 Da that is 44% identical over a region of 264 amino acids to a product of unknown function encoded by an open reading frame associated with mycolic acid synthesis in Mycobacterium tuberculosis H37Ra. Extracts of Escherichia coli cells expressing the acr1 gene catalyzed the reduction of acyl-CoA to the corresponding fatty aldehyde, indicating that the gene encodes a novel fatty acyl-CoA reductase.
Identification and characterization of pbpA encoding Bacillus subtilis penicillin-binding protein 2A
Authors: Murray T , Popham D L , Setlow P
Published: November 2016

Amino acid sequence analysis of tryptic peptides derived from purified penicillin-binding protein PBP2a of Bacillus subtilis identified the coding gene (now termed pbpA) as yqgF, which had been sequenced as part of the B. subtilis genome project; pbpA encodes a 716-residue protein with sequence similarity to class B high-molecular-weight PBPs. Use of a pbpA-lacZ fusion showed that pbpA was expressed predominantly during vegetative growth, and the transcription start site was mapped using primer extension analysis. Insertional mutagenesis of pbpA resulted in no changes in the growth rate or morphology of vegetative cells, in the ability to produce heat-resistant spores, or in the ability to trigger spore germination when compared to the wild type. However, pbpA spores were unable to efficiently elongate into cylindrical cells and were delayed significantly in spore outgrowth. This provides evidence that PBP2a is involved in the synthesis of peptidoglycan associated with cell wall elongation in B. subtilis.
In vivo construction of a hybrid pathway for metabolism of 4-nitrotoluene in Pseudomonas fluorescens
Authors: Michán C , Delgado A , Haïdour A , Lucchesi G , Ramos J L
Published: November 2016

Pseudomonas fluorescens 410PR grows on 4-nitrobenzoate but does not metabolize 4-nitrotoluene. The TOL pWW0 delta pm plasmid converts 4-nitrotoluene into 4-nitrobenzoate through its upper pathway, but it does not metabolize 4-nitrobenzoate. P. fluorescens 410PR(pWW0 delta pm) transconjugants were isolated and found to be able to grow on 4-nitrotoluene. This phenotype was stable after growth for at least 300 generations without any selective pressure. P. fluorescens 410PR(pWW0 delta pm) converted 4-nitrotoluene into 4-nitrobenzoate via 4-nitrobenzylalcohol and 4-nitrobenzaldehyde. 4-Nitrobenzoate was metabolized via 4-hydroxylaminobenzoate and finally yielded NH4+ and 3,4-dihydroxybenzoate, which was mineralized.
Heme compounds as iron sources for nonpathogenic Rhizobium bacteria
Authors: Noya F , Arias A , Fabiano E
Published: November 2016

Many animal-pathogenic bacteria can use heme compounds as iron sources. Like these microorganisms, rhizobium strains interact with host organisms where heme compounds are available. Results presented in this paper indicate that the use of hemoglobin as an iron source is not restricted to animal-pathogenic microorganisms. We also demonstrate that heme, hemoglobin, and leghemoglobin can act as iron sources under iron-depleted conditions for Rhizobium meliloti 242. Analysis of iron acquisition mutant strains indicates that siderophore-, heme-, hemoglobin-, and leghemoglobin-mediated iron transport systems expressed by R. meliloti 242 share at least one component.
Pericardiectomy as a diagnostic and therapeutic procedure
Authors: Konik Ewa , Geske Jeffrey , Edwards William , Gersh Bernard
Published: November 2016

A 70-year-old man presented with recent onset, predominantly right-sided heart failure. Echocardiogram demonstrated features of hypertensive heart disease and was suggestive of, but non-diagnostic for, constrictive pericarditis (CP). CT demonstrated mild pericardial thickening. Right heart catheterisation showed elevation and equalisation of diastolic pressures in all cardiac chambers with early rapid filling, minimal ventricular interdependence, and no dissociation of intrathoracic and intracardiac pressures. While several features pointed towards CP, the minimal ventricular interdependence and no dissociation of intrathoracic and intracardiac pressures suggested other pathology. Diagnostic pericardiectomy was performed, after which the central venous pressure decreased from 22 to 12 mm Hg. Pathology revealed pericardial fibrosis. The patient experienced sustained resolution of his heart failure. A potential explanation for lack of CP criteria was the presence of hypertensive heart disease. CP needs to be considered when approaching patients with heart failure as diagnostic evaluation can be multifaceted and treatment curative.
One-year outcome after Descemet membrane endothelial keratoplasty (DMEK) comparing sulfur hexafluoride (SF6) 20% versus 100% air for anterior chamber tamponade
Authors: Schaub Friederike , Enders Philip , Snijders Katharina , Schrittenlocher Silvia , Siebelmann Sebastian , Heindl Ludwig M , Bachmann Björn O , Cursiefen Claus
Published: November 2016

Purpose

To investigate 1-year clinical outcome and complication rates following Descemet membrane endothelial keratoplasty (DMEK) with sulfur hexafluoride 20% (SF620%) anterior chamber tamponade compared with conventionally used 100% air for primary graft attachment during DMEK surgery.

Methods

Records of 1112 consecutive DMEKs were reviewed retrospectively and grouped by anterior chamber tamponade used during DMEK surgery (SF620% vs 100% air). Outcome measures included intraocular pressure (IOP), best spectacle-corrected visual acuity (BSCVA), endothelial cell density (ECD) and central corneal thickness (CCT) at 1, 3, 6 and 12 months after DMEK surgery. Complication rates were assessed, including intraoperative and postoperative complications, and graft detachment rate requiring rebubbling.

Results

A total of 854 cases were included in this study. In 105 cases (12.3%), DMEK was performed with SF620%, and in 749 cases (87.7%) 100% air was used for anterior chamber tamponade. Outcome results for IOP, BSCVA, ECD and CCT at all follow-up time points were comparable for both anterior chamber tamponade groups without statistical significant differences (p≥0.05), but graft detachment rate requiring rebubbling was significantly lower in the SF620% group (p<0.001).

Conclusion

Whereas SF620% anterior chamber tamponade does not seem to negatively affect the clinical outcome of DMEK surgery within the first postoperative year, use of SF620% significantly reduces the rate of rebubblings.
Rethinking the cost of healthcare in low-resource settings: the value of time-driven activity-based costing
Authors: McBain Ryan K , Jerome Gregory , Warsh Jonathan , Browning Micaela , Mistry Bipin , Faure Peterson Abnis I , Pierre Claire , Fang Anna P , Mugunga Jean Claude , Rhatigan Joseph , Leandre Fernet , Kaplan Robert
Published: November 2016

Low-income and middle-income countries account for over 80% of the world's infectious disease burden, but <20% of global expenditures on health. In this context, judicious resource allocation can mean the difference between life and death, not just for individual patients, but entire patient populations. Understanding the cost of healthcare delivery is a prerequisite for allocating health resources, such as staff and medicines, in a way that is effective, efficient, just and fair. Nevertheless, health costs are often poorly understood, undermining effectiveness and efficiency of service delivery. We outline shortcomings, and consequences, of common approaches to estimating the cost of healthcare in low-resource settings, as well as advantages of a newly introduced approach in healthcare known as time-driven activity-based costing (TDABC). TDABC is a patient-centred approach to cost analysis, meaning that it begins by studying the flow of individual patients through the health system, and measuring the human, equipment and facility resources used to treat the patients. The benefits of this approach are numerous: fewer assumptions need to be made, heterogeneity in expenditures can be studied, service delivery can be modelled and streamlined and stronger linkages can be established between resource allocation and health outcomes. TDABC has demonstrated significant benefits for improving health service delivery in high-income countries but has yet to be adopted in resource-limited settings. We provide an illustrative case study of its application throughout a network of hospitals in Haiti, as well as a simplified framework for policymakers to apply this approach in low-resource settings around the world.
Hospital-diagnosed atopic dermatitis and long-term risk of myocardial infarction: a population-based follow-up study
Authors: Riis Jette Lindorff , Vestergaard Christian , Hjuler Kasper Fjellhaugen , Iversen Lars , Jakobsen Lars , Deleuran Mette S , Olsen Morten
Published: November 2016

Objective

Atopic dermatitis (AD) is an inflammatory skin disorder with a childhood prevalence reaching 20%. An estimated 50% of patients have a life-long chronic course. The purpose of this study was to estimate the risk of first-time myocardial infarction (MI) in patients with AD compared with a general population cohort.

Design

Cohort study.

Setting

Denmark.

Participants

Using population-based medical registries, we identified individuals born in Denmark from 1947 to 1983 with at least two hospital-diagnoses of AD following inpatient admissions or hospital-based outpatient visits at any age from 1977 to 2013. Individuals with AD were matched with general population controls (10:1) for birth-year and gender. Unique personal identifiers permitted unambiguous data linkage.

Primary outcome measures

Follow-up began on the date of AD diagnosis (index date for general population controls) and continued until death, emigration, MI or the year 2013. We computed the 15-year-cumulative incidence of MI following a diagnosis of AD. Comparing patients with AD with the general population cohort, we computed HRs of MI presented with 95% CIs and adjusted for history of diabetes mellitus, hypertension, hyperlipidaemia or stroke, educational level, birth-year and sex.

Results

We identified 4814 patients diagnosed with AD. The cumulative incidence of MI was 0.6% for patients with AD and 0.4% for their matched controls. The corresponding adjusted HR was 1.74 (1.21 to 2.49). The HR for patients who were not in need of systemic treatment was 1.58 (1.02 to 2.45) and it was 2.40 (1.27 to 4.45) for those who were treated with azathioprine, methotrexate or cyclosporine.

Conclusions

Hospital-diagnosed AD was associated with increased risk of MI compared with the general population.
Patient safety events in out-of-hospital paediatric airway management: a medical record review by the CSI-EMS
Authors: Hansen Matthew , Meckler Garth , Lambert William , Dickinson Caitlin , Dickinson Kathryn , Van Otterloo Joshua , Guise Jeanne-Marie
Published: November 2016

Objective

To describe the frequency and characterise the nature of patient safety events in paediatric out-of-hospital airway management.

Methods

We conducted a retrospective cross-sectional medical record review of all ‘lights and sirens’ emergency medicine services transports from 2008 to 2011 in patients <18 years of age in the Portland Oregon metropolitan area. A chart review tool (see online supplementary appendix) was adapted from landmark patient safety studies and revised after pilot testing. Expert panels of physicians and paramedics performed blinded reviews of each chart, identified safety events and described their nature. The primary outcomes were presence and severity of patient safety events related to airway management including oxygen administration, bag-valve-mask ventilation (BVM), airway adjuncts and endotracheal intubation (ETI).

supplementary appendix

Results

From the 11 328 paediatric transports during the study period, there were 497 ‘lights and sirens’ (code 3) transports (4.4%). 7 transports were excluded due to missing data. Of the 490 transports included in the analysis, 329 had a total of 338 airway management procedures (some had more than 1 procedure): 61.6% were treated with oxygen, 15.3% with BVM, 8.6% with ETI and 2% with airway adjuncts. The frequency of errors was: 21% (71/338) related to oxygen use, 9.8% (33/338) related to BVM, 9.5% (32/338) related to intubation and 0.9% (3/338) related to airway adjunct use. 58% of intubations required 3 or more attempts or failed altogether. Cardiac arrest was associated with higher odds of a severe error.

Conclusions

Errors in paediatric out-of-hospital airway management are common, especially in the context of intubations and during cardiac arrest.
Concerns, perceived need and competing priorities: a qualitative exploration of decision-making and non-participation in a population-based flexible sigmoidoscopy screening programme to prevent colorectal cancer
Authors: Hall N , Birt L , Rees C J , Walter F M , Elliot S , Ritchie M , Weller D , Rubin G
Published: November 2016

Objective

Optimising uptake of colorectal cancer (CRC) screening is important to achieve projected health outcomes. Population-based screening by flexible sigmoidoscopy (FS) was introduced in England in 2013 (NHS Bowel scope screening). Little is known about reactions to the invitation to participate in FS screening, as offered within the context of the Bowel scope programme. We aimed to investigate responses to the screening invitation to inform understanding of decision-making, particularly in relation to non-participation in screening.

Design

Qualitative analysis of semistructured in-depth interviews and written accounts.

Participants and setting

People from 31 general practices in the North East and East of England invited to attend FS screening as part of NHS Bowel scope screening programme were sent invitations to take part in the study. We purposively sampled interviewees to ensure a range of accounts in terms of beliefs, screening attendance, sex and geographical location.

Results

20 screeners and 25 non-screeners were interviewed. Written responses describing reasons for, and circumstances surrounding, non-participation from a further 28 non-screeners were included in the analysis. Thematic analysis identified a range of reactions to the screening invitation, decision-making processes and barriers to participation. These include a perceived or actual lack of need; inability to attend; anxiety and fear about bowel preparation, procedures or hospital; inability or reluctance to self-administer an enema; beliefs about low susceptibility to bowel cancer or treatment and understanding of harm and benefits. The strength, rather than presence, of concerns about the test and perceived need for reassurance were important in the decision to participate for screeners and non-screeners. Decision-making occurs within the context of previous experiences and day-to-day life.

Conclusions

Understanding the reasons for non-participation in FS screening can help inform strategies to improve uptake and may be transferable to other screening programmes.
Gabapentinoids for chronic low back pain: a protocol for systematic review and meta-analysis of randomised controlled trials
Authors: Shanthanna Harsha , Gilron Ian , Thabane Lehana , Devereaux Philip J , Bhandari Mohit , AlAmri Rizq , Rajarathinam Manikandan , Kamath Sriganesh
Published: November 2016

Introduction

Chronic low back pain (CLBP) is a common condition and causes significant pain, distress and disability across the world. It is multifactorial in aetiology and is challenging to manage. Although the underlying mechanism of pain is predominantly non-specific, many argue that there is a substantial neuropathic pain element. Neuropathic pain is more severe, with significant disability. Gabapentinoids, including gabapentin and pregabalin, have proven efficacy in some neuropathic pain conditions. Despite no clear evidence, a substantial population of patients with CLBP are treated with gabapentinoids.

Objectives

We aim to assess whether the use of gabapentinoids is effective and safe in the treatment of predominant CLBP, by conducting a systematic review and meta-analysis of randomised control trials (RCTs).

Methodology

We will search the databases of MEDLINE, EMBASE and Cochrane for RCTs published in English language and have used gabapentinoids for the treatment of CLBP. Study selection and data extraction will be performed independently by paired reviewers using structured electronic forms, piloted between pairs of reviewers. The review outcomes will be guided by Initiative on Methods, Measurement and Pain Assessment in Clinical Trials guidelines, with pain relief as the primary outcome. We propose to carry out meta-analysis if there are three or more studies in a particular outcome domain, using a random effects model. Pooled outcomes will be reported as weighted mean differences or standardised mean differences and risk ratios with their corresponding 95% CIs, for continuous outcomes and dichotomous outcomes, respectively. Rating of quality of evidence will be reported using GRADE summary of findings table.

Discussion

The proposed systematic review will be able to provide valuable evidence to help decision-making in the use of gabapentinoids for the treatment of CLBP. This will help advance patient care and potentially highlight limitations in existing evidence to direct future research.

Ethics and dissemination

Being a systematic review, this study would not necessitate ethical review and approval. We plan to report and publish our study findings in a high impact medical journal, with online access.

Trial registration number

CRD42016034040.
Sugar, dental caries and the incidence of acute rheumatic fever: a cohort study of Māori and Pacific children
Authors: Thornley Simon , Marshall Roger J , Bach Katie , Koopu Pauline , Reynolds Gary , Sundborn Gerhard , Ei Win Le Shwe Sin
Published: November 2016

Objective

To determine whether dental caries, as an indicator of cumulative exposure to sugar, is associated with the incidence of acute rheumatic fever and chronic rheumatic heart disease, in Māori and Pacific children aged 5 and 6 years at their first dental visit.

Materials and methods

A cohort study was undertaken which linked school dental service records of caries with national hospital discharge and mortality records. Cox models were used to investigate the strength of the association between dental caries and rheumatic fever incidence.

Results

A total of 20 333 children who were free of rheumatic heart disease at enrolment were available for analysis. During a mean follow-up time of 5 years, 96 children developed acute rheumatic fever or chronic rheumatic heart disease. After adjustment for potential confounders, children with five or more primary teeth affected by caries were 57% (95% CI: 20% to 106%) more likely to develop disease during follow-up, compared to children whose primary teeth were caries free. The population attributable to the risk for caries in this cohort was 22%.

Conclusions

Dental caries is positively associated with the incidence of acute rheumatic fever and chronic rheumatic heart disease in Māori and Pacific children. Sugar intake, an important risk factor for dental caries, is also likely to influence the aetiology of rheumatic fever.
L Ron Hubbard's science fiction quest against psychiatry
Authors: Hirshbein Laura
Published: November 2016

Layfayette Ronald Hubbard (1911–1986) was a colourful and prolific American writer of science fiction in the 1930s and 1940s. During the time between his two decades of productivity and his return to science fiction in 1980, Hubbard founded the Church of Scientology. In addition to its controversial status as a religion and its troubling pattern of intimidation and litigation directed towards its foes, Scientology is well known as an organised opponent to psychiatry. This paper looks at Hubbard's science fiction work to help understand the evolution of Scientology's antipsychiatry stance, as well as the alternative to psychiatry offered by Hubbard.
The real-world use of regorafenib for metastatic colorectal cancer: multicentre analysis of treatment pattern and outcomes in Hong Kong
Authors: Lam Ka-On , Lee Kin-Chung , Chiu Joanne , Lee Victor Ho-Fun , Leung Roland , Choy T S , Yau Thomas
Published: November 2016

Purpose of the study

To evaluate the benefits and tolerability of regorafenib in the real-world setting, we performed a multicentre analysis in Hong Kong.

Study design

Individual patient data were retrieved from three leading oncology centres in Hong Kong for analyses. All patients with metastatic colorectal cancer (mCRC) treated with regorafenib after failure of all standard systemic options were included.

Results

From July 2013 to December 2015, 45 consecutive patients treated with regorafenib for mCRC were analysed. The median age was 63. Twenty patients were started at 160 mg, while the other 25 patients were started at a lower dose. The median progression-free survival was 15.6 weeks (95% CI 13.1 to 18.1 weeks) and the median overall survival was 30.4 weeks (95% CI 16.6 to 44.3 weeks). Among the 31 evaluable patients, only 1 patient (3.2%) achieved partial response and another 10 patients (32.3%) had stable disease. The commonest grade 3 non-haematological adverse event (AE) was hand–foot skin reaction (26.7%) and the commonest grade 3 or 4 haematological AE was anaemia (8.9%). Notably, patients who were started on a lower dose of regorafenib had significantly lower risk of grade 3 treatment-emergent AEs. Overall, 78.3% of the patients had dose reduction during the first and second cycles. Patients older than 65 years were more likely to experience cycle suspension and require dose reduction.

Conclusions

Our study confirmed the efficacy and tolerability of regorafenib in the real-world setting. It also suggested that individualised dosing of regorafenib in patients with mCRC might result in better clinical outcomes.
What is the reliability of non-trained investigators in recognising structural MRI lesions of sacroiliac joints in patients with recent inflammatory back pain? Results of the DESIR cohort
Authors: Jacquemin Charlotte , Rubio Vargas Roxana , van den Berg Rosaline , Thévenin Fabrice , Lenczner Gregory , Reijnierse Monique , Ferkal Salah , Le Corvoisier Philippe , Rahmouni Alain , Loeuille Damien , Feydy Antoine , Dougados Maxime , van der Heijde Désirée , Claudepierre Pascal
Published: November 2016

Objective

The objective of this study was to evaluate the reliability of recognising structural lesions on MRI (erosions, fatty lesions, ankylosis) of the sacroiliac joints (MRI-SIJ) in clinical practice compared to a central reading in patients with a possible recent axial spondyloarthritis (axSpA).

Methods

Patients aged 18–50 years, with recent (<3 years) and chronic (≥3 months) inflammatory back pain, suggestive of axSpA were included in the DEvenir des Spondyloarthrites Indifférenciées Récentes (DESIR) cohort. MRI-SIJ structural lesions were scored by non-trained local readers, and by two trained central readers. Local readers scored each SIJ as normal, doubtful or definite lesions. Central readers scored separately each type of lesion. The central reading (mean of the two central readers’ scores) was the external standard. Agreement (κ) was calculated first between local (3 definitions of a positive MRI-SIJ) and central readings (9 definitions), and then between the two central readers.

Results

664/708 patients with complete available images were included. Agreements between local and central readings were overall ‘fair’, except when considering at least 2 or 3 fatty lesions and at least 3 erosions and/or fatty lesions where agreement was ‘moderate’. Agreement between central readers was similar. MRI-SIJ was positive for 52.6% of patients according to central reading (at least 1 structural lesion) and for 35.4% of patients according to local reading (at least unilateral ‘doubtful‘ or ‘definite’ structural lesions).

Conclusions

Agreement on a positive structural MRI-SIJ was fair to moderate between local and central readings, as well as between central readers. The reliability improved when fatty lesions were considered.

Trial registration number

NCTO 164 8907.
Spaces and places: an historical overview of the development of geology in Austria (Habsburg Monarchy) in the eighteenth and nineteenth centuries
Authors: Klemun Marianne
Published: November 2016

Geology as a discipline emerged in Austria between the eighteenth and nineteenth centuries from mineralogy and geognosy, two sciences dominated by descriptive rather than theoretical approaches. Austria, in this context, refers to the conglomerate of different states under the Habsburg Monarchy that existed until 1918. Two key aspects shaped the development of geology and the establishment of public institutions: mining industries and natural history collections. As a result of their close relation, the k.k. Geologische Reichsanstalt (Imperial Geological Survey) was founded in Vienna in 1849, which had repercussions for the entire empire. The Reichsanstalt carried out a geological survey of all the territories of the Habsburg Monarchy, resulting in a unifying geological map published in 1867. Geology was triggered and accelerated not least by the industrialization processes of the nineteenth century. From the 1850s, geology no longer focused only on the empire and its provinces, but took a global perspective. From 1862 a growing number of university chairs for geology were established, making the ‘new’ academic discipline visible. The internationally outstanding research work of Eduard Suess contributed to this success story.
History of a discussion: selected aspects of the Earth expansion v. plate tectonics theories
Authors: Cwojdzinski Stefan
Published: November 2016

A global tensional system of mid-ocean ridges was discovered at the end of the 1950s. All the world's oceans were proved to be young Mesozoic–Cenozoic structures and these new ideas in geotectonics led directly to the theories of ocean floor spreading and plate tectonics. The development of the idea of ocean floor spreading proved to be a crucial turning point in modern geotectonics. These facts could suggest the expansion of the Earth. However, the view that invoked Wegener's theory of continental drift – plate tectonics – which assumed a constant size for the Earth, remained the most prevalent idea. From this point of view, plate tectonics is just a hypothesis for a non-expanding Earth. The followers of plate tectonics believe that the drift of the continents relies on their movement upon the asthenosphere. New oceanic crust has to be consumed in the so-called subduction zones, compensating for the growth of the Earth. If the process of the opening of the oceans is a unidirectional global process – and it is not compensated for – then the Earth will increase in size. The process of the widening of new oceans, such as the Atlantic, Arctic and Indian oceans, should be simultaneous with the shrinking of the Palaeo-Pacific Ocean. If the Pacific Ocean expands, then the expansion of the Earth is inevitable.
New Biofunctional Loading of Natural Antimicrobial Agent in Biodegradable Polymeric Films for Biomedical Applications
Authors: Ghafoor Bakhtawar , Ali Murtaza Najabat , Ansari Umar , Bhatti Muhammad Faraz , Mir Mariam , Akhtar Hafsah , Darakhshan Fatima
Published: November 2016

The study focuses on the development of novelAloe verabased polymeric composite films and antimicrobial suture coatings. Polyvinyl alcohol (PVA), a synthetic biocompatible and biodegradable polymer, was combined withAloe vera, a natural herb used for soothing burning effects and cosmetic purposes. The properties of these two materials were combined together to get additional benefits such as wound healing and prevention of surgical site infections. PVA andAloe verawere mixed in a fixed quantity to produce polymer based films. The films were screened for antibacterial and antifungal activity against bacterial (E. coli,P. aeruginosa) and fungal strains (Aspergillus flavusandAspergillus tubingensis) screened.Aloe verabased PVA films showed antimicrobial activity against all the strains; the lowestAloe veraconcentration (5%) showed the highest activity against all the strains.In vitrodegradation and release profile of these films was also evaluated. The coating for sutures was prepared,in vitroantibacterial tests of these coated sutures were carried out, and later onin vivostudies of these coated sutures were also performed. The results showed that sutures coated withAloe vera/PVA coating solution have antibacterial effects and thus have the potential to be used in the prevention of surgical site infections andAloe vera/PVA based films have the potential to be used for wound healing purposes.
Prospective, Multicenter Clinical Trial of Everolimus as Primary Therapy in Waldenstrom Macroglobulinemia (WMCTG 09-214)
Authors: Treon Steven P. , Meid Kirsten , Tripsas Christina , Heffner Leonard T. , Eradat Herbert , Badros Ashraf Z. , Xu Lian , Hunter Zachary R. , Yang Guang , Patterson Christopher J. , Gustine Joshua , Castillo Jorge J. , Matous Jeffrey , Ghobrial Irene M.
Published: November 2016

Purpose: Everolimus inhibits mTOR, a component of PI3K/AKT prosurvival signaling triggered by MYD88 and CXCR4-activating mutations in Waldenstrom macroglobulinemia.

Experimental design: We evaluated everolimus in a prospective, multicenter study of 33 symptomatic, previously untreated Waldenstrom macroglobulinemia patients. Intended therapy consisted of everolimus (10 mg/day) until progression or unacceptable toxicity. Dose deescalation was permitted. The study was registered at www.clinicaltrials.gov (NCT00976248).

Results: At best response, median serum IgM levels declined from 4,440 to 1,360 mg/dL (P < 0.0001), median hemoglobin rose from 10.8 to 12 g/dL (P = 0.001), and median bone marrow disease burden declined from 75% to 52.5% in serially biopsied patients. The ORR and major response rates were 72.7% and 60.6%, respectively. Among genotyped patients, nonresponders associated with wild-type MYD88 and mutated CXCR4 status. Median time to response was 4 weeks. Discordance between serum IgM levels and bone marrow disease burden was remarkable. With a median follow-up of 13.1 (range, 1.6–64.6 months), the median time to progression was 21 months for all patients and 33 months for major responders. Discontinuation of everolimus led to rapid serum IgM rebound in 7 patients and symptomatic hyperviscosity in 2 patients. Toxicity led to treatment discontinuation in 27% of patients, including 18% for pneumonitis.

Conclusions: Everolimus is active in previously untreated Waldenstrom macroglobulinemia. IgM discordance is common, and treatment cessation can often lead to rapid serum IgM rebound. Pneumonitis also appears more pronounced in untreated versus previously treated Waldenstrom macroglobulinemia patients. The risks and benefits of everolimus should be carefully weighed against other primary Waldenstrom macroglobulinemia therapy options. Clin Cancer Res; 23(10); 2400–4. ©2016 AACR.