bluuebunny/crossref_metadata_2025 huggingface dataset:
Suitability of the forearm for non‐invasive blood pressure measurement in children
Authors: Greaney David , Nakhjavani Solmaz , Desmond Fiona , Tormey Mark , Dawkins Ian , O'Hare Brendan
Published: October 2017

Background

Measurement of forearm blood pressure (BP) in pediatric patients during general anesthesia is periodically employed despite a lack of evidence for this practice. Upper arm BP measurement may be impossible to perform for either patient or surgical reasons, and the forearm has theoretical benefits over the lower leg when an alternate site is required. We hypothesize that forearm BP measurement provides an accurate and reliable alternative to the upper arm. Published adult data do not support this hypothesis, and the little pediatric data published contain methodological shortcomings.

Methods

A dedicated, externally calibrated noninvasive oscillometer was used to compare BP measurements in the upper arm and ipsilateral forearm of pediatric patients undergoing general anesthesia prior to application of a surgical stimulus. Both upper arm BP and ipsilateral forearm BP were sequentially measured 20 seconds apart on 3 separate occasions with an appropriately sized cuff. The systolic, diastolic, and mean blood pressures were recorded under steady‐state conditions.

Results

Thirty‐five elective surgical patients aged 1 to 10 years were studied. The bias (±limits of agreement) for forearm minus upper arm blood pressures were as follows: mean BP −1.3 mm Hg (±7.2), diastolic BP −3.3 (±5.3), and systolic BP +3.2 mm Hg (±8.3). Differences greater than ±5 mm Hg occurred in 59% (systolic BP), 42% (diastolic BP), and 46% (mean BP) of all observations and greater than ±10 mm Hg in 17% (systolic BP), 8.6% (diastolic BP), and 15% (mean BP).

Conclusion

The differences within mean ±1.96 standard deviations reside considerably outside the clinically accepted tolerance of ±5 mm Hg. Thus, the forearm may not be used interchangeably with upper limb BP readings in anesthetized healthy children. Future use of the forearm for BP measurement requires a validated anthropomorphically appropriate forearm cuff.
Responses of the soil fungal communities to the co‐invasion of two invasive species with different cover classes
Authors: Wang C. , Zhou J. , Liu J. , Jiang K. , Xiao H. , Du D.
Published: October 2017

Soil fungal communities play an important role in the successful invasion of non‐native species. It is common for two or more invasive plant species to co‐occur in invaded ecosystems.

This study aimed to determine the effects of co‐invasion of two invasive species (Erigeron annuus and Solidago canadensis) with different cover classes on soil fungal communities using high‐throughput sequencing.

Invasion of E. annuus and/or S. canadensis had positive effects on the sequence number, operational taxonomic unit (OTU) richness, Shannon diversity, abundance‐based cover estimator (ACE index) and Chao1 index of soil fungal communities, but negative effects on the Simpson index. Thus, invasion of E. annuus and/or S. canadensis could increase diversity and richness of soil fungal communities but decrease dominance of some members of these communities, in part to facilitate plant further invasion, because high soil microbial diversity could increase soil functions and plant nutrient acquisition. Some soil fungal species grow well, whereas others tend to extinction after non‐native plant invasion with increasing invasion degree and presumably time. The sequence number, OTU richness, Shannon diversity, ACE index and Chao1 index of soil fungal communities were higher under co‐invasion of E. annuus and S. canadensis than under independent invasion of either individual species.

The co‐invasion of the two invasive species had a positive synergistic effect on diversity and abundance of soil fungal communities, partly to build a soil microenvironment to enhance competitiveness of the invaders. The changed diversity and community under co‐invasion could modify resource availability and niche differentiation within the soil fungal communities, mediated by differences in leaf litter quality and quantity, which can support different fungal/microbial species in the soil.
Identification of sesame (Sesamum indicum L.) chromosomes using the BAC‐FISH system
Authors: Zhao R. , Miao H. , Song W. , Chen C. , Zhang H.
Published: October 2017

Sesame (Sesamum indicum L.; Pedaliaceae) is a commercially valuable oilseed crop with high oil content. Its small genome size favours the genomic analysis of key biological processes, such as oil synthesis and metabolism. However, the 13 chromosome pairs of sesame have not been characterised because of technological limitations and their small size.

We constructed a BAC library comprising 57,600 BAC clones for sesame. The estimated genome coverage of the sesame BAC library was 13.8×. The successive double colour fluorescence in situ hybridisation (FISH) with bacterial artificial chromosomes (BACs) for sesame was established in this study.

Subsequently, the 13 sesame chromosome pairs were individually differentiated using 17 specific BACs for the first time. The schematic of the sesame chromosome set was drawn according to the chromosome relative length and relative position of the BAC signal. The cytogenetic characteristics of sesame chromosomes were also explored.

The results provide the technical background required for further cytogenetic map construction, genome assembly and localisation of the DNA sequence in sesame.
The novel allele, HLA‐A*31:108, identified by sequence‐based typing in a Chinese individual
Authors: Ding H. , Huang Y. , Chen Y. , Ye X.
Published: October 2017

The HLA‐A*31:108 allele was found.
An Arabidopsis divergent pumilio protein, APUM24, is essential for embryogenesis and required for faithful pre‐rRNA processing
Authors: Shanmugam Thiruvenkadam , Abbasi Nazia , Kim Hyung‐Sae , Kim Ho Bang , Park Nam‐il , Park Guen Tae , Oh Sung Aeong , Park Soon Ki , Muench Douglas G. , Choi Yeonhee , Park Youn‐Il , Choi Sang‐Bong
Published: October 2017

Pumilio RNA‐binding proteins are largely involved in mRNA degradation and translation repression. However, a few evolutionarily divergent Pumilios are also responsible for proper pre‐rRNA processing in human and yeast. Here, we describe an essential Arabidopsis nucleolar Pumilio, APUM24, that is expressed in tissues undergoing rapid proliferation and cell division. A T‐DNA insertion for APUM24 did not affect the male and female gametogenesis, but instead resulted in a negative female gametophytic effect on zygotic cell division immediately after fertilization. Additionally, the mutant embryos displayed defects in cell patterning from pro‐embryo through globular stages. The mutant embryos were marked by altered auxin maxima, which were substantiated by the mislocalization of PIN1 and PIN7 transporters in the defective embryos. Homozygous apum24 callus accumulates rRNA processing intermediates, including uridylated and adenylated 5.8S and 25S rRNA precursors. An RNA–protein interaction assay showed that the histidine‐tagged recombinant APUM24 binds RNAin vitro with no apparent specificity. Overall, our results demonstrated that APUM24 is required for rRNA processing and early embryogenesis in Arabidopsis.
Genome‐wide characterization of differential transcript usage in Arabidopsis thaliana
Authors: Vaneechoutte Dries , Estrada April R. , Lin Ying‐Chen , Loraine Ann E. , Vandepoele Klaas
Published: October 2017

Alternative splicing and the usage of alternate transcription start‐ or stop sites allows a single gene to produce multiple transcript isoforms. Most plant genes express certain isoforms at a significantly higher level than others, but under specific conditions this expression dominance can change, resulting in a different set of dominant isoforms. These events of differential transcript usage (DTU) have been observed for thousands of Arabidopsis thaliana, Zea mays and Vitis vinifera genes, and have been linked to development and stress response. However, neither the characteristics of these genes, nor the implications of DTU on their protein coding sequences or functions, are currently well understood. Here we present a dataset of isoform dominance and DTU for all genes in the AtRTD2 reference transcriptome based on a protocol that was benchmarked on simulated data and validated through comparison with a published reverse transciptase‐polymerase chain reaction panel. We report DTU events for 8148 genes across 206 public RNA‐Seq samples, and find that protein sequences are affected in 22% of the cases. The observed DTU events show high consistency across replicates, and reveal reproducible patterns in response to treatment and development. We also demonstrate that genes with different evolutionary ages, expression breadths and functions show large differences in the frequency at which they undergo DTU, and in the effect that these events have on their protein sequences. Finally, we showcase how the generated dataset can be used to explore DTU events for genes of interest or to find genes with specific DTU in samples of interest.
Fetal/neonatal alloimmune thrombocytopenia due to anti‐CD36 antibodies: antibody evaluations by CD36‐transfected cell lines
Authors: Lin Marie , Xu Xiuzhang , Lee Hui‐Lin , Liang Der‐Cheng , Santoso Sentot
Published: October 2017

BACKGROUND

Isoantibodies against CD36 (platelet glycoprotein 4), developed in Type I CD36‐deficient mothers are frequently reported as the cause of fetal/neonatal alloimmune thrombocytopenia in the Asian population. Therefore, further detailed characterization of anti‐CD36–mediated fetal/neonatal alloimmune thrombocytopenia is warranted. Here, we report the characterization of a patient with fetal/neonatal alloimmune thrombocytopenia in a Taiwanese family caused by anti‐CD36 isoantibodies using a novel antigen‐capture method.

STUDY DESIGN AND METHODS

Platelets and monocytes were analyzed for CD36 expression by flow cytometry. Sequencing analysis of the CD36 gene was performed to identify the mutation underlying the CD36 deficiency. Stable transfected human embryonic kidney HEK293 cells expressing recombinant CD36 were established. These cells were used for the characterization of anti‐CD36 isoantibodies by flow cytometry, immunoprecipitation, and antigen‐capture assay.

RESULTS

Flow cytometry analysis revealed a total absence of CD36 on both platelets and monocytes of the mother (Type I CD36‐deficient) caused by heterozygous deletions of the CD36 gene (332_333delCA and c.1254 + 6_1254 + 11delTATTTG). Analysis of maternal serum with CD36‐transfected HEK293 cells by flow cytometry, immunoprecipitation, and antigen‐capture assay demonstrated the presence of anti‐CD36 isoantibodies in maternal serum. Interestingly, this antibody could not be detected by the monoclonal antibody immobilization of platelet antigens assay when anti‐CD36 monoclonal antibody (clone FA6‐152) was used as the capture antibody.

CONCLUSION

This case reemphasizes the role of anti‐CD36 isoantibodies on the pathomechanism of fetal/neonatal alloimmune thrombocytopenia. The fact that the monoclonal antibody immobilization of platelet antigens assay does not seem to be reliable for the identification of all anti‐CD36 antibodies indicates that screening of anti‐CD36 isoantibodies by a monoclonal antibody‐independent method, as presented here, should be considered.
Text Messages Promoting Mental Imagery Increase Self-Reported Physical Activity in Older Adults: A Randomized Controlled Study
Authors: Robin Nicolas , Toussaint Lucette , Coudevylle Guillaume R. , Ruart Shelly , Hue Olivier , Sinnapah Stephane
Published: October 2017

Objective:This study tested whether text messages prompting adults 50 years of age and older to perform mental imagery would increase aerobic physical activity (APA) duration using a randomized parallel trial design.Method:Participants were assigned to an Imagery 1, Imagery 2, or placebo group. For 4 weeks, each group was exposed to two conditions (morning text message vs. no morning text message). In the morning message condition, the imagery groups received a text message with the instruction to mentally imagine performing an APA, and the placebo group received a placebo message. All participants received an evening text message of “Did you do your cardio today? If yes, what did you do?” for 3 days per week.Results:Participants of the imagery groups reported significantly more weekly minutes of APA in the morning text message condition compared with the no morning message condition.Conclusion:Electronic messages were effective at increasing minutes of APA.
Classical strong metal–support interactions between gold nanoparticles and titanium dioxide
Authors: Tang Hailian , Su Yang , Zhang Bingsen , Lee Adam F. , Isaacs Mark A. , Wilson Karen , Li Lin , Ren Yuegong , Huang Jiahui , Haruta Masatake , Qiao Botao , Liu Xin , Jin Changzi , Su Dangsheng , Wang Junhu , Zhang Tao
Published: October 2017

The classical strong metal–support interaction between TiO2and IB metals was demonstrated.
Integrating networks and comparative genomics reveals retroelement proliferation dynamics in hominid genomes
Authors: Levy Orr , Knisbacher Binyamin A. , Levanon Erez Y. , Havlin Shlomo
Published: October 2017

Integrating comparative genomics and network science delineates the dynamics of retroelement activity in the genome
Lead halide perovskites: Crystal-liquid duality, phonon glass electron crystals, and large polaron formation
Authors: Miyata Kiyoshi , Atallah Timothy L. , Zhu X.-Y.
Published: October 2017

Efficient dynamic screening is key to the success of lead halide perovskites in solar cells and light emitting devices.
Enzymatic synthesis of core 2 O-glycans governs the tissue-trafficking potential of memory CD8
+
T cells
Authors: Osborn Jossef F. , Mooster Jana L. , Hobbs Samuel J. , Munks Michael W. , Barry Conrad , Harty John T. , Hill Ann B. , Nolz Jeffrey C.
Published: October 2017

Central memory CD8
+
T cells are the major memory CD8
+
T cell subset that traffics into inflamed, nonlymphoid tissues.
An A/U-Rich Enhancer Region Is Required for High-Level Protein Secretion through the HlyA Type I Secretion System
Authors: Khosa Sakshi , Scholz Romy , Schwarz Christian , Trilling Mirko , Hengel Hartmut , Jaeger Karl-Erich , Smits Sander H. J. , Schmitt Lutz
Published: October 2017

Efficient protein secretion is often a valuable alternative to classic cellular expression to obtain homogenous protein samples. Early on, bacterial type I secretion systems (T1SS) were employed to allow heterologous secretion of fusion proteins. However, this approach was not fully exploited, as many proteins could not be secreted at all or only at low levels. Here, we present an engineered microbial secretion system which allows the effective production of proteins up to a molecular mass of 88 kDa. This system is based on the hemolysin A (HlyA) T1SS of the Gram-negative bacterium
Escherichia coli
, which exports polypeptides when fused to a hemolysin secretion signal. We identified an A/U-rich enhancer region upstream of
hlyA
required for effective expression and secretion of selected heterologous proteins irrespective of their prokaryotic, viral, or eukaryotic origin. We further demonstrate that the ribosomal protein S1 binds to the
hlyA
A/U-rich enhancer region and that this region is involved in the high yields of secretion of functional proteins, like maltose-binding protein or human interferon alpha-2.

IMPORTANCE
A 5′ untranslated region of the mRNA of substrates of type I secretion systems (T1SS) drastically enhanced the secretion efficiency of the endogenously secreted protein. The identification of ribosomal protein S1 as the interaction partner of this 5′ untranslated region provides a rationale for the enhancement. This strategy furthermore can be transferred to fusion proteins allowing a broader, and eventually a more general, application of this system for secreting heterologous fusion proteins.
Housefly (Musca domestica) and Blow Fly (Protophormia terraenovae) as Vectors of Bacteria Carrying Colistin Resistance Genes
Authors: Zhang Jilei , Wang Jiawei , Chen Li , Yassin Afrah Kamal , Kelly Patrick , Butaye Patrick , Li Jing , Gong Jiansen , Cattley Russell , Qi Kezong , Wang Chengming
Published: October 2017

Flies have the capacity to transfer pathogens between different environments, acting as one of the most important vectors of human diseases worldwide. In this study, we trapped flies on a university campus and tested them for mobile resistance genes against colistin, a last-resort antibiotic in human medicine for treating clinical infections caused by multidrug-resistant Gram-negative bacteria. Quantitative PCR assays we developed showed that 34.1% of
Musca domestica
(86/252) and 51.1% of
Protophormia terraenovae
(23/45) isolates were positive for the
mcr-1
gene, 1.2% of
M. domestica
(3/252) and 2.2% of
P. terraenovae
(2.2%, 1/45) isolates were positive for
mcr-2
, and 5.2% of
M. domestica
(13/252) and 44.4% of
P. terraenovae
(20/45) isolates were positive for
mcr-3
. Overall, 4.8% (9/189) of bacteria isolated from the flies were positive for the
mcr-1
gene (
Escherichia coli
: 8.3%, 4/48;
Enterobacter cloacae
: 12.5%, 1/8;
Providencia alcalifaciens
: 11.8%, 2/17;
Providencia stuartii
: 4.9%, 2/41), while none were positive for
mcr-2
and
mcr-3
. Four
mcr-1
-positive isolates (two
P. stuartii
and two
P. alcalifaciens
) from blow flies trapped near a dumpster had a MIC for colistin above 4 mg/ml. This study reports
mcr-1
carriage in
Providencia
spp. and detection of
mcr-2
and
mcr-3
after their initial identification in Belgium and China, respectively. This study suggests that flies might contribute significantly to the dissemination of bacteria, carrying these genes into a large variety of ecological niches. Further studies are warranted to explore the roles that flies might play in the spread of colistin resistance genes.

IMPORTANCE
Antimicrobial resistance is recognized as one of the most serious global threats to human health. An option for treatment of the Gram-negative ESKAPE (
Enterococcus faecium
,
Staphylococcus aureus
,
Klebsiella pneumoniae
,
Acinetobacter baumannii
,
Pseudomonas aeruginosa
, and
Enterobacter
species) bacteria with multiple drug resistance was the reintroduction of the older antibiotic colistin. However, a mobile colistin resistance gene (
mcr-1
) has recently been found to occur widely; very recently, two other colistin resistance genes (
mcr-2
and
mcr-3
) have been identified in Belgium and China, respectively. In this study, we report the presence of colistin resistance genes in flies. This study also reports the carriage of colistin resistance genes in the genus
Providencia
and detection of
mcr-2
and
mcr-3
after their initial identification. This study will stimulate more in-depth studies to fully elucidate the transmission mechanisms of the colistin resistance genes and their interaction.
Methanogenic Paraffin Biodegradation: Alkylsuccinate Synthase Gene Quantification and Dicarboxylic Acid Production
Authors: Oberding Lisa K. , Gieg Lisa M.
Published: October 2017

Paraffinic
n
-alkanes (>C
17
) that are solid at ambient temperature comprise a large fraction of many crude oils. The comparatively low water solubility and reactivity of these long-chain alkanes can lead to their persistence in the environment following fuel spills and pose serious problems for crude oil recovery operations by clogging oil production wells. However, the degradation of waxy paraffins under the anoxic conditions characterizing contaminated groundwater environments and deep subsurface energy reservoirs is poorly understood. Here, we assessed the ability of a methanogenic culture enriched from freshwater fuel-contaminated aquifer sediments to biodegrade the model paraffin
n
-octacosane (C
28
H
58
). Compared with that in controls, the consumption of
n
-octacosane was coupled to methane production, demonstrating its biodegradation under these conditions.
Smithella
was postulated to be an important C
28
H
58
degrader in the culture on the basis of its high relative abundance as determined by 16S rRNA gene sequencing. An identified
assA
gene (known to encode the α subunit of alkylsuccinate synthase) aligned most closely with those from other
Smithella
organisms. Quantitative PCR (qPCR) and reverse transcription qPCR assays for
assA
demonstrated significant increases in the abundance and expression of this gene in C
28
H
58
-degrading cultures compared with that in controls, suggesting
n
-octacosane activation by fumarate addition. A metabolite analysis revealed the presence of several long-chain α,ω-dicarboxylic acids only in the C
28
H
58
-degrading cultures, a novel observation providing clues as to how methanogenic consortia access waxy hydrocarbons. The results of this study broaden our understanding of how waxy paraffins can be biodegraded in anoxic environments with an application toward bioremediation and improved oil recovery.

IMPORTANCE
Understanding the methanogenic biodegradation of different classes of hydrocarbons has important applications for effective fuel-contaminated site remediation and for improved recovery from oil reservoirs. Previous studies have clearly demonstrated that short-chain alkanes (<C
17
) can be biodegraded anaerobically, but less is understood regarding the biodegradation of longer chain waxy alkanes (>C
17
) that comprise many fuel mixtures. Using an enrichment culture derived from a freshwater fuel-contaminated site, we demonstrate that the model waxy alkane
n
-octacosane can be biodegraded under methanogenic conditions by a presumed
Smithella
phylotype. Compared with that of controls, we show an increased abundance and expression of the
assA
gene, which is known to be important for anaerobic
n
-alkane metabolism. Metabolite analyses revealed the presence of a range of α,ω-dicarboxylic acids found only in
n
-octacosane-degrading cultures, a novel finding that lends insight as to how anaerobic communities may access waxes as growth substrates in anoxic environments.
Plantaricyclin A, a Novel Circular Bacteriocin Produced by Lactobacillus plantarum NI326: Purification, Characterization, and Heterologous Production
Authors: Borrero Juan , Kelly Eoin , O'Connor Paula M. , Kelleher Philip , Scully Colm , Cotter Paul D. , Mahony Jennifer , van Sinderen Douwe
Published: October 2017

Bacteriocins from lactic acid bacteria (LAB) are of increasing interest in recent years due to their potential as natural preservatives against food and beverage spoilage microorganisms. In a screening study for LAB, we isolated from olives a strain,
Lactobacillus plantarum
NI326, with activity against the beverage-spoilage bacterium
Alicyclobacillus acidoterrestris
. Genome sequencing of NI326 enabled the identification of a gene cluster (designated
plc
) encoding a putative circular bacteriocin and proteins involved in its modification, transport, and immunity. This novel bacteriocin, named plantaricyclin A (PlcA), was grouped into the circular bacteriocin subgroup II due to its high degree of similarity with other gassericin A-like bacteriocins. Purification of PlcA from the supernatant of
Lb. plantarum
NI326 resulted in an active peptide with a molecular mass of 5,570 Da, corresponding to that predicted from the (processed) PlcA amino acid sequence. The
plc
gene cluster was cloned and expressed in
Lactococcus lactis
NZ9000, resulting in the production of an active 5,570-Da bacteriocin in the supernatant. PlcA is believed to be produced as a 91-amino-acid precursor with a 33-amino-acid leader peptide, which is predicted to be removed, followed by joining of the N and C termini via a covalent linkage to form the mature 58-amino-acid circular bacteriocin PlcA. We report the characterization of a circular bacteriocin produced by
Lb. plantarum
. The inhibition displayed against
A. acidoterrestris
highlights its potential use as a preservative in food and beverages.

IMPORTANCE
In this work, we describe the purification and characterization of an antimicrobial peptide, termed plantaricyclin A (PlcA), produced by a
Lactobacillus plantarum
strain isolated from olives. This peptide has a circular structure, and all genes involved in its production, circularization, and secretion were identified. PlcA shows antimicrobial activity against different strains, including
Alicyclobacillus acidoterrestris
, a common spoilage bacterium, which causes substantial economic losses in the beverage industry every year. In this study, we describe a circular antimicrobial peptide, PlcA, for a
Lactobacillus plantarum
strain.
Use of Comparative Genomics-Based Markers for Discrimination of Host Specificity in Fusarium oxysporum
Authors: van Dam Peter , de Sain Mara , ter Horst Anneliek , van der Gragt Michelle , Rep Martijn
Published: October 2017

The polyphyletic nature of manyformae specialesofFusarium oxysporumprevents molecular identification of newly encountered strains based on conserved, vertically inherited genes. Alternative molecular detection methods that could replace labor- and time-intensive disease assays are therefore highly desired. Effectors are functional elements in the pathogen-host interaction and have been found to show very limited sequence diversity between strains of the sameforma specialis, which makes them potential markers for host-specific pathogenicity. We therefore compared candidate effector genes extracted from 60 existing and 22 newly generated genome assemblies, specifically targeting strains affecting cucurbit plant species. Based on these candidate effector genes, a total of 18 PCR primer pairs were designed to discriminate between each of the seven Cucurbitaceae-affectingformae speciales. When tested on a collection of strains encompassing different clonal lineages of theseformae speciales, nonpathogenic strains, and strains of otherformae speciales, they allowed clear recognition of the host range of each evaluated strain. WithinFusarium oxysporumf. sp.melonismore genetic variability exists than anticipated, resulting in threeF. oxysporumf. sp.melonismarker patterns that partially overlapped with the cucurbit-infectingFusarium oxysporumf. sp.cucumerinum,Fusarium oxysporumf. sp.niveum,Fusarium oxysporumf. sp.momordicae, and/orFusarium oxysporumf. sp.lagenariae. ForF. oxysporumf. sp.niveum, a multiplex TaqMan assay was evaluated and was shown to allow quantitative and specific detection of template DNA quantities as low as 2.5 pg. These results provide ready-to-use marker sequences for the mentionedF. oxysporumpathogens. Additionally, the method can be applied to find markers distinguishing other host-specific forms ofF. oxysporum.

IMPORTANCEPathogenic strains ofFusarium oxysporumare differentiated intoformae specialesbased on their host range, which is normally restricted to only one or a few plant species. However, horizontal gene transfer between strains in the species complex has resulted in a polyphyletic origin of host specificity in many of theseformae speciales. This hinders accurate and rapid pathogen detection through molecular methods. In our research, we compared the genomes of 88 strains ofF. oxysporumwith each other, specifically targeting virulence-related genes that are typically highly similar within eachforma specialis. Using this approach, we identified marker sequences that allow the discrimination ofF. oxysporumstrains affecting various cucurbit plant species through different PCR-based methods.
Novel Metabolic Pathway for
N
-Methylpyrrolidone Degradation in Alicycliphilus sp. Strain BQ1
Authors: Solís-González Claudia Julieta , Domínguez-Malfavón Lilianha , Vargas-Suárez Martín , Gaytán Itzel , Cevallos Miguel Ángel , Lozano Luis , Cruz-Gómez M. Javier , Loza-Tavera Herminia
Published: October 2017

The molecular mechanisms underlying the biodegradation of
N
-methylpyrrolidone (NMP), a widely used industrial solvent that produces skin irritation in humans and is teratogenic in rats, are unknown.
Alicycliphilus
sp. strain BQ1 degrades NMP. By studying a transposon-tagged mutant unable to degrade NMP, we identified a six-gene cluster (
nmpABCDEF
) that is transcribed as a polycistronic mRNA and encodes enzymes involved in NMP biodegradation.
nmpA
and the transposon-affected gene
nmpB
encode an
N
-methylhydantoin amidohydrolase that transforms NMP to γ-
N
-methylaminobutyric acid; this is metabolized by an amino acid oxidase (NMPC), either by demethylation to produce γ-aminobutyric acid (GABA) or by deamination to produce succinate semialdehyde (SSA). If GABA is produced, the activity of a GABA aminotransferase (GABA-AT), not encoded in the
nmp
gene cluster, is needed to generate SSA. SSA is transformed by a succinate semialdehyde dehydrogenase (SSDH) (NMPF) to succinate, which enters the Krebs cycle. The abilities to consume NMP and to utilize it for growth were complemented in the transposon-tagged mutant by use of the
nmpABCD
genes. Similarly,
Escherichia coli
MG1655, which has two SSDHs but is unable to grow in NMP, acquired these abilities after functional complementation with these genes. In wild-type (wt) BQ1 cells growing in NMP, GABA was not detected, but SSA was present at double the amount found in cells growing in Luria-Bertani medium (LB), suggesting that GABA is not an intermediate in this pathway. Moreover,
E. coli
GABA-AT deletion mutants complemented with
nmpABCD
genes retained the ability to grow in NMP, supporting the possibility that γ-
N
-methylaminobutyric acid is deaminated to SSA instead of being demethylated to GABA.

IMPORTANCE
N
-Methylpyrrolidone is a cyclic amide reported to be biodegradable. However, the metabolic pathway and enzymatic activities for degrading NMP are unknown. By developing molecular biology techniques for
Alicycliphilus
sp. strain BQ1, an environmental bacterium able to grow in NMP, we identified a six-gene cluster encoding enzymatic activities involved in NMP degradation. These findings set the basis for the study of new enzymatic activities and for the development of biotechnological processes with potential applications in bioremediation.
Improving the ability to review preoperative radiographs intraoperatively in trauma and orthopaedic theatres at Lancashire teaching hospitals
Authors: Jump Christopher
Published: October 2017

Background

The ability to review preoperative radiographs during trauma and orthopaedic surgery is essential for the surgeon to provide optimum treatment to the patient. However, due to current information technology (IT) systems, screen-savers frequently interrupt the ability to review images and theatre staff are not routinely available to deactivate the screen-saver. This prolongs theatre time for the patient and affects the quality of care provided. The aim of this quality improvement project was to improve the availability of radiographs for the surgeon to review intraoperatively.

Method/results

Data were collected from all trauma and orthopaedic theatres at two hospital sites covering all subspecialties and including emergency and elective cases. Baseline measurements showed that the frequency of preoperative radiographs not interrupted during an operation was 0% (0/50). Following this the Trust’s IT systems were improved to prevent activation of the screen-saver on the theatre computers using the generic theatre login details. After the first-cycle intervention, data were collected showing 52% (14/27) of preoperative radiographs were not interrupted by a screen-saver. The cause for this result being less than expected was investigated and found to be due to an alternative computer login being used on the theatre computers at one of the hospital sites. Education of theatre staff was then undertaken to ensure the correct theatre login was used and notices to remind staff placed on the theatre computers. After the second-cycle intervention, data were collected showing that 100% (26/26) of preoperative radiographs were not interrupted during operative time allowing the surgeon to review images when required.

Conclusion/implications

This quality improvement project has made changes to theatre IT systems and practices of theatre staff which has resulted in a significant improvement in the ability for the operating surgeon to review preoperative radiographs intraoperatively.
Long-term outcomes of patients with pulmonary arteriovenous malformations considered for lung transplantation, compared with similarly hypoxaemic cohorts
Authors: Shovlin Claire L , Buscarini Elisabetta , Hughes J Michael B , Allison David J , Jackson James E
Published: October 2017

Introduction

Pulmonary arteriovenous malformations (PAVMs) may not be amenable to treatment by embolisation or surgical resection, and many patients are left with significant hypoxaemia. Lung transplantation has been undertaken. There is no guidance on selection criteria.

Methods

To guide transplantation listing assessments, the outcomes of the six patients who had been considered for transplantation were compared with a similarly hypoxaemic patient group recruited prospectively between 2005 and 2016 at the same UK institution.

Results

Six patients had been formally considered for lung transplantation purely for PAVMs. One underwent a single lung transplantation for diffuse PAVMs and died within 4 weeks of surgery. The other five were not transplanted, in four cases at the patients’ request. Their current survival ranges from 16 to 27 (median 21) years post-transplant assessment.

Of 444 consecutive patients with PAVMs recruited between 2005 and 2016, 42 were similarly hypoxaemic to the ‘transplant-considered’ cohort (SaO2<86.5%). Hypoxaemic cohorts maintained arterial oxygen content (CaO2) through secondary erythrocytosis and higher haemoglobin. The ‘transplant-considered’ cohort had similar CaO2to the hypoxaemic comparator group, but higher Medical Research Council (MRC) dyspnoea scores (p=0.023), higher rates of cerebral abscesses (p=0.0043) and higher rates of venous thromboemboli (p=0.0009) that were evident before and after the decision to list for transplantation.

Conclusions

The non-transplanted patients demonstrated marked longevity. Symptoms and comorbidities were better predictors of health than oxygen measurements. While a case-by-case decision, weighing survival estimates and quality of life will help patients in their decision making, the data suggest a very strong case must be made before lung transplantation is considered.
Burden of bacterial upper respiratory tract pathogens in school children of Nepal
Authors: Thapa Sangita , Gokhale Shishir , Sharma Annavarapu Laxminarasimha , Sapkota Lokendra Bahadur , Ansari Shamshul , Gautam Rajendra , Shrestha Sony , Neopane Puja
Published: October 2017

Introduction

Respiratory tract infections are one of the most common human infections in all age group and important cause of mortality and morbidity worldwide. Most bacterial upper respiratory tract infections are vaccine preventable. This study aimed to determine the prevalence of carrier state of bacterial upper respiratory tract pathogens among school children. It also aimed to study their antibiograms.

Methods

The specimen from posterior pharyngeal wall and tonsils were collected from 204 participants on calcium alginate coated swabs (HiMedia). Isolates were identified by standard microbiological methods and tested for in vitro antibiotic susceptibility testing by modified Kirby-Bauer disc diffusion method.

Results

In this study,Streptococcus pneumoniae(16.6%) was the most common bacterial pathogen recovered, followed byStaphylococcus aureus(14.7%), β-haemolytic streptococci (non-Group A) (8.8%),Streptococcus pyogenes(5.3%) andCorynebacterium diphtheriae(3.4%). The Gram negative bacteria wereKlebsiella pneumoniae(4.9%),Haemophilus influenzae(3.4%) andNeisseria meningitidis(1.4%). Important findings in antibiogram include high resistance ofStreptococcus pneumoniaeto penicillin (91.17%) and resistance ofS. aureusto oxacillin (23.3%).

Conclusion

Pharyngeal colonisation byS. pneumoniaewas found high among school children and this calls for an urgent need to include pneumococcal vaccine in routine national immunisation schedule of Nepal given the high burden of invasive pneumococcal disease. Despite expected universal vaccination, pharyngeal colonisation byC. diphtheriaeis possible and there is possibility of transmission of these respiratory pathogens to other healthy children.
Alcohol drinking and cigarette smoking in relation to risk of active tuberculosis: prospective cohort study
Authors: Soh Avril Zixin , Chee Cynthia Bin Eng , Wang Yee-Tang , Yuan Jian-Min , Koh Woon-Puay
Published: October 2017

Introduction

Heavy alcohol consumption increases the risk of active tuberculosis (TB). However, the relation between lower levels of alcohol intake and TB risk remains unclear. We aimed to evaluate the association between alcohol intake and risk of active TB and assess whether the associations were modified by smoking status, which is another risk factor for active TB.

Methods

The Singapore Chinese Health Study is a prospective cohort of 63 257 adults aged 45–74 years recruited from 1993 to 1998. Information on alcohol intake and smoking history was collected at recruitment. Active TB cases were identified via linkage with National TB Notification Registry.

Results

During a mean follow-up of 16.8 years, 1249 incident cases of active TB were identified. Among non-smokers, compared with total abstinence, participants who had monthly to weekly intake of alcohol had reduced TB risk (HR 0.70, 95% CI 0.55 to 0.89), but this reduction in risk with low-dose drinking was not observed among current smokers (HR 0.96, 95% CI 0.77 to 1.18; p for interaction=0.02). Comparatively, drinking 2+ drinks daily was associated with increased TB risk among current smokers (HR 1.51, 95% CI 1.11 to 2.05). This increased risk was not observed among non-smokers (HR 0.93, 95% CI 0.49 to 1.77) and the interaction between alcohol intake and smoking status was of borderline significance (p for interaction=0.08). In joint effect, compared with those who neither smoked nor drank, the risk of active TB increased from 1.82 (95% CI 1.57 to 2.10) in current smokers who were non-drinkers to 3.16 (95% CI 2.35 to 4.24) in current smokers who also drank 2+ drinks daily.

Conclusion

While low intake of alcohol may protect against active TB among non-smokers, drinking 2+ drinks daily could act synergistically with smoking to increase the risk of active TB in current smokers.
New biomarker for acute ischaemic stroke: plasma glycogen phosphorylase isoenzyme BB
Authors: Park Kwang-Yeol , Ay Ilknur , Avery Ross , Caceres Juan Alfredo , Siket Matthew S , Pontes-Neto Octavio M , Zheng Hui , Rost Natalia S , Furie Karen L , Sorensen Alma Gregory , Koroshetz Walter J , Ay Hakan
Published: October 2017

Background

Glycogen phosphorylase is the key enzyme that breaks down glycogen to yield glucose-1-phosphate in order to restore depleted energy stores during cerebral ischaemia. We sought to determine whether plasma levels of glycogen phosphorylase BB (GPBB) isoform increased in patients with acute ischaemic stroke (AIS).

Methods

We studied plasma GPBB levels within 12 hours and again at 48±24 hours of symptom onset in 172 patients with imaging-confirmed AIS and 133 stroke-free individuals. We determined the ability of plasma GPBB to discriminate between cases and controls and examined the predictive value of plasma GPBB for 90-day functional outcome, 90-day survival and acute lesion volumes on neuroimaging.

Results

The mean (SD) GPBB levels were higher in cases (46.3±38.6 ng/mL at first measurement and 38.6±36.5 ng/mL at second measurement) than in controls (4.1±7.6 ng/mL, p<0.01 for both). The area under the receiver operating characteristic (ROC) curve for case–control discrimination based on first GPBB measurement was 0.96 (95% CI 0.93 to 0.98). The sensitivity and specificity based on optimal operating point on the ROC curve (7.0 ng/mL) were both 93%. GPBB levels increased in 90% of patients with punctate infarcts (<1.5 mL) and in all patients admitted within the first 4.5 hours of onset. There was no correlation between GPBB concentration and either clinical outcome or acute infarct volume.

Conclusion

GPBB demonstrates robust response to acute ischaemia and high sensitivity for small infarcts. If confirmed in more diverse populations that also include stroke mimics, GPBB could find utility as a stand-alone marker for acute brain ischaemia.
Short delay to initiate plasma exchange is the strongest predictor of outcome in severe attacks of NMO spectrum disorders
Authors: Bonnan Mickael , Valentino Rudy , Debeugny Stéphane , Merle Harold , Fergé Jean-Louis , Mehdaoui Hossein , Cabre Philippe
Published: October 2017

Introduction

Severe attacks of neuromyelitis optica spectrum disorder (NMO-SD) are improved by plasma exchange (PLEX) given as an adjunctive therapy. Initial studies failed to demonstrate a delay of PLEX treatment influenced clinical outcome; however PLEX was always used late. We examine the clinical consequences of delay in PLEX initiation on severe optic neuritis and spinal cord attacks in NMO-SD.

Methods

All of our patients who suffered attacks of NMO-SD, treated in our centre by PLEX, were retrospectively considered for inclusion. Primary outcome was defined as complete improvement. Secondary poor/good outcomes were respectively defined to be the higher/lower third of Delta-Expanded Disability Status Scale (EDSS) (late minus baseline EDSS). Delays from clinical onset to PLEX initiation were categorised for multivariate analysis.

Results

Of the 60 patients included, NMO-SD criteria (2015) were fulfilled in 92%. One hundred and fifteen attacks were included and received PLEX with a median of 7 days (0–54) after clinical onset. The probability to regain complete improvement continuously decreased from 50% for PLEX given at day 0 to 1%–5% after day 20. Through multivariate analysis, the baseline impairment and PLEX delay were associated with the probability to complete improvement (OR 5.3; 95% CI 1.8 to 15.9). Reducing the PLEX delay also influenced the good secondary outcome but not the poor secondary outcome.

Conclusions

These results confirm an improved clinical benefit of early initiation of PLEX during severe attacks of NMO-SD. Perceiving PLEX as a rescue therapy only after steroid failure could be deleterious.
Serotonin transporter gene polymorphisms may be associated with poststroke neurological recovery after escitalopram use
Authors: Lee Eun-Jae , Oh Mi-Sun , Kim Jong S , Chang Dae-Il , Park Jong-Ho , Cha Jae-Kwan , Heo Ji Hoe , Sohn Sung-Il , Kim Dong-Eog , Kim Hahn Young , Kim Jei , Seo Woo-Keun , Lee Jun , Park Sang-Won , Kim Yun Joong , Lee Byung-Chul
Published: October 2017

Objective

Selective serotonin reuptake inhibitors (SSRIs) putatively improve neurological recovery after stroke. We aimed to investigate whether serotonin transporter (SERT) gene polymorphisms are related to the responsiveness to SSRIs in the poststroke neurological recovery.

Methods

This was a post hoc analysis of the EMOTION study (ClinicalTrials.gov NCT01278498), a randomised, placebo-controlled, double-blind trial examining the efficacy of escitalopram on emotional and neurological disturbances after acute stroke. Patients with no/minimal disability initially (modified Rankin Scale (mRS) 0–1) were excluded. Of the participants, 301 underwent genetic studies of the STin2 (a variable number tandem repeat (VNTR) in intron 2) (STin2 12/10 and STin2 12/12 genotypes) and 5-HTTLPR (a variable-length repeat in the promoter region) polymorphisms of SERT. We explored whether neurological function (National Institutes of Health Stroke Scale (NIHSS) score and mRS) at 3 months would differ according to SERT polymorphisms within each treatment arm (escitalopram and placebo).

Results

Among the escitalopram users (n=159), neurological function in subjects with STin2 12/10 (n=29) improved significantly more than that in STin2 12/12 carriers (n=130) at 3 months. After adjusting for age, initial NIHSS and depression, STin2 12/10 independently predicted a good clinical outcome (mRS 0–1) (OR 2.99, 95% CI 1.04 to 8.58) at 3 months. However, differences between STin2 polymorphisms were not shown in the placebo group (n=142). 5-HTTLPR polymorphisms were not associated with neurological recovery in any treatment group.

Conclusion

STin2 VNTR polymorphisms may be associated with poststroke neurological recovery after SSRI therapy. Further studies are needed to identify the role of serotonin in neurological recovery after stroke.
Shame and the vulnerable self in medical contexts: the compassionate solution
Authors: Gilbert Paul
Published: October 2017

Shame is a powerful experience that plays a vital role in a whole range of aspects of the clinical encounter. Shame experiences can have an impact on our psychological and physiological state and on how we experience ourselves, others and our relationships. The medical encounter is an obvious arena for shame because we are presenting (aspects of) our bodies and minds that can be seen as unattractive and undesirable, diseased, decayed and injured with the various excretions that typically might invite disgust. In contrast, experiences of compassion of acceptance, validation and kindness and can increase approach, openness and preparedness to engage with painful difficult scenarios. While shame is an experience that separates, segregates, marginalises and disengages people, caring and compassion facilitate integration, (re)connection and support. Given the potential opposite impacts of these different types of social experience, this paper will outline their evolutionary origins and compare and contrast them with particular reference to the medical context.
Anaesthetising a cat with tetralogy of Fallot for non‐cardiac surgery
Authors: Pekkola Vuokko , Braun Christina , Larenza Menzies Maria Paula
Published: October 2017

Tetralogy of Fallot (ToF) is a rare congenital cardiac disease, which has been described in many species, including cats. It often causes cyanosis due to deoxygenated blood entering the systemic circulation. The prognosis of uncorrected ToF is generally poor, but some individuals with mild forms of the disease can survive beyond three years of age. A cat with mild, asymptomatic ToF was anaesthetised for elective ovariectomy. The cat was premedicated intramuscularly with methadone and alfaxalone. Anaesthesia was induced with alfaxalone and midazolam and maintained with sevoflurane in oxygen using mechanical ventilation. Intraoperative analgesia was provided with a constant rate infusion of fentanyl and local anaesthesia. Phenylephrine was infused during anaesthesia to maintain adequate systemic vascular resistance. Mild bradycardia during ovarian tissue handling was treated successfully with atropine. Meloxicam and buprenorphine were used to provide postoperative analgesia. The cat did not experience major complications during anaesthesia and made a full recovery.
Epidural anaesthesia for treatment of neuropathic pain associated with pelvic limb amputation in a domestic shorthair cat
Authors: Primason Lily Knox , Gleed Robin , Boesch Jordyn M
Published: October 2017

An approximately 3‐year‐old, intact female, domestic shorthair cat presented to a shelter with a chronic fracture of the right femur. She underwent amputation of her right pelvic limb at the coxofemoral joint. On adoption two weeks following amputation, her owner noted clinical signs of pain associated with the stump, including resentment of stump palpation, paroxysmal vocalization and excessive grooming of the stump. Physical examination, including orthopaedic and neurological evaluation, was unremarkable except for subtle resentment of stump palpation. The history and clinical signs were consistent with neuropathic pain associated with the stump. A multimodal analgesic protocol consisting of oral meloxicam, gabapentin and amitriptyline was initiated; however, only mild improvement was noted and these medications were discontinued. A lumbosacral epidural injection of bupivacaine, buprenorphine and triamcinolone was performed under fluoroscopic guidance. The epidural injection resulted in immediate, complete resolution of clinical signs of pain that had not returned nine months later.