bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Auricular Acupressure Therapy on Primary Dysmenorrhea for Female High School Students in South Korea
Authors: Cha Nam Hyun , Sok Sohyune R.
Published: August 2016

Purpose

To examine the effect of auricular acupressure therapy on primary dysmenorrhea among female high school students in South Korea.

Design

A randomized controlled trial was employed.

Methods

The study sample consisted of 91 female high school students, with 45 participants in the experimental group and 46 in the control group in two regions of South Korea. The average age of the participants was 16.7 years, and the average age of menarche was 12.2 years. Auricular acupressure therapy including an auricular acupressure needle on skin paper tape was applied on an ear for 3 days during periods of extreme primary dysmenorrhea. The acupoint names were Jagung, Sinmun, Gyogam, and Naebunbi. For the placebo control group, only the skin paper tape without an auricular acupressure needle was applied on the same acupoints. Measures used were the Menstrual Distress Questionnaire to assess primary dysmenorrhea, and the visual analog scale to assess abdominal and back pain of participants.

Findings

There were significant differences on abdominal pain (t = 24.594, p < .001), back pain (t = 22.661, p < .001), and primary dysmenorrhea (t = 32.187, p < .001) between the two groups. Auricular acupressure therapy decreased abdominal pain, back pain, and primary dysmenorrhea of female high school students in South Korea.

Conclusions

Auricular acupressure therapy was an effective intervention for alleviating abdominal pain, back pain, and primary dysmenorrhea of female high school students in South Korea. For feasibility of the auricular acupressure therapy in practice, it is needed to train and learn the exact positions of acupoints in ear.

Clinical Relevance

Health providers should consider providing auricular acupressure therapy as an alternative method for reducing abdominal and back pain, and primary dysmenorrhea in female high school students in South Korea.
Delirium the under‐recognised syndrome: survey of healthcare professionals’ awareness and practice in the intensive care units
Authors: Selim Abeer A , Ely E. Wesley
Published: August 2016

Aims and objectives

To survey intensive care unit healthcare professionals’ awareness and practice related to delirium.

Background

Despite the current evidence revealing the risks linked to delirium and advances in practice guidelines promoting delirium assessment, healthcare professionals show little sensitivity towards delirium and evident training needs.

Design

The study had a cross‐sectional survey design.

Methods

A sample of 168 intensive care unit healthcare professionals including nurses and physicians completed a semistructured questionnaire to survey their awareness, screening and management of delirium in intensive care units. The survey took place at 11 intensive care units from academic (university) and nonacademic (nonuniversity) governmental hospitals in Mansoura, Egypt.

Results

The mean score of delirium awareness was 64·4 ± 14·0 among intensive care unit healthcare professionals. Awareness of delirium was significantly lower when definition of delirium was not provided, among diploma nurses compared to bachelor degree nurses and physicians, among those who did not attend any workshop/lecture or read an article related to delirium and lastly, those who work in an intensive care unit when <50% of patients develop delirium. The survey found that only 26·8% of the healthcare professionals screen for delirium on a routine basis, and 14·3% reported attending workshops or lectures or reading an article related to delirium in the last year. In screening delirium, healthcare professionals did not use any tools, nor did they follow adopted protocols or guidelines to manage delirium. To manage delirium, 52·4% of the participants reported using sedatives, 36·9% used no drugs, and 10·7% reported using antipsychotics (primarily haloperidol).

Conclusion

Intensive care unit healthcare professionals do not have adequate training or routine screening of delirium. There is an evident absence of using standardised tools or adapting protocols to monitor and manage delirium.

Relevance to clinical practice

This study has the potentials to shed some lights on the variables that might explain the problem of underdiagnosing delirium by healthcare professionals at intensive care units in Mansoura.
Newborn hearing screening in Queensland 2009–2011: Comparison of hearing screening and diagnostic audiological assessment between term and preterm infants
Authors: Calcutt Trent L , Dornan Dimity , Beswick Rachael , Tudehope David I
Published: August 2016

Aim

This study compares rates and timing of newborn hearing screening outcomes, audiological assessment and hearing loss diagnosis between infants of different gestational age groups. Early identification and management of sensorineural hearing loss (SNHL), ideally by 3–6 months of age, facilitates speech and language optimisation. Literature stratifying hearing screening and diagnostic audiology assessment by gestational age groups is lacking.

Methods

Subjects were infants with recorded gestational ages receiving newborn hearing screening in Queensland between 2009 and 2011. Data were provided through the Queensland Healthy Hearing database. Infants were analysed in <34 weeks, 34–36+6 weeks, 37–38+6 weeks and ≥39 weeks gestational age groups.

Results

Infants (175 911) were eligible for analysis, 7.9% being preterm. Per 1000 infants analysed, bilateral SNHL of >40 dB occurred in 2.4 for <34, 1.4 for 34–36+6, 0.7 for 37–38+6 and 0.7 for ≥39 weeks gestation. Diagnoses attributable to newborn hearing screening direct referral were 93.1% for bilateral >40 dB SNHL and 88.2% for other hearing loss. Relative to term, preterm infants had a higher incidence of direct and targeted surveillance referrals, audiology assessment and hearing loss diagnosis. Preterm infants were screened later after birth.

Conclusions

Specific hearing screening and diagnosis characteristics differed between preterm infants <34 and 34–36+6 weeks gestation, and term infants. Consideration of unique gestational age strata characteristics supports care individualisation. Preterm infants represent a diagnostic challenge, with higher rates of bilateral >40 dB SNHL than term but correspondingly higher false positive results on screening, justifying vigilant monitoring. Focused research into specific risk factors in preterm infants is warranted.
Immune cells in repair of the infarcted myocardium
Authors: Chen Bijun , Frangogiannis Nikolaos G.
Published: August 2016

The immune system plays a critical role in both repair and remodeling of the infarcted myocardium. Danger signals released by dying cardiomyocytes mobilize, recruit, and activate immune cells, triggering an inflammatory reaction. CXC chemokines containing the ELR motif attract neutrophils, while CC chemokines mediate recruitment of mononuclear cell subpopulations, contributing to clearance of the infarct from dead cells and matrix debris. Immune cell subsets also participate in suppression and containment of the postinfarction inflammatory response by secreting anti‐inflammatory mediators, such as IL‐10 and TGF‐β. As proinflammatory signaling is suppressed, macrophage subpopulations, mast cells and lymphocytes, activate fibrogenic and angiogenic responses, contributing to scar formation. In the viable remodeling myocardium, chronic activation of immune cells may promote fibrosis and hypertrophy. This review discusses the role of immune cells in repair and remodeling of the infarcted myocardium. Understanding the role of immune cells in myocardial infarction is critical for the development of therapeutic strategies aimed at protecting the infarcted heart from adverse remodeling. Moreover, modulation of immune cell phenotype may be required in order to achieve the visionary goal of myocardial regeneration.
Successful switching of anticoagulants in a patient with superficial siderosis and atrial fibrillation
Authors: Inatomi Yuichiro , Nakajima Makoto , Yonehara Toshiro , Ando Yukio
Published: August 2016

A 78‐year‐old man with atrial fibrillation treated with warfarin experienced acute headache, deafness, cerebellar ataxia and dysuria. Superficial siderosis was diagnosed on brain imaging; however, no bleeding sources were detected. His neurological deficits and bloody cerebrospinal fluid diminished after withdrawal of warfarin. Apixaban was started 2 months after the onset, and his deficits and siderosis had not recurred 18 months after switching these anticoagulants.
Determinants and co‐expression of anti‐predator responses in amphibian tadpoles: a meta‐analysis
Authors: Hossie Thomas , Landolt Kristen , Murray Dennis L.
Published: August 2016

A wide range of taxa respond to perceived predation risk (PPR) through inducible defenses, and many prey are capable of responding both behaviorally and morphologically to the same risk event. In cases where multiple defenses confer protection by independent means (i.e. they are mechanistically independent) responses will either be co‐expressed, or the expression of one defense will limit the capacity (or need) to respond along another axis. Our ability to generate a broad understanding of these patters has been limited, in part, by difficulties in comparing results across studies that employ distinct experimental protocols. Using the extensive literature on tadpole responses to PPR, we conducted a meta‐analysis to identify the ecological and experimental determinants of inducible defence expression. We then assessed whether the magnitude of response to PPR along behavioural versus morphological response axes was positively, or negatively, correlated. The most commonly quantified responses to perceived risk in tadpoles included reductions in movement and swimming behaviour, and altered tail morphology. Our analyses reveal that tadpole behavioural responses are strongly influenced by prey family, predator taxon, evolutionary history with the predator (native versus non‐native), amount of prey consumed by the predator, and how perceived risk was manipulated (e.g. presence versus absence of alarm cues). Tail morphology was similarly influenced by these factors, but also whether the target prey was palatable to predators. Thus, our results identify ecological and experimental features that critically influence the observed effect size in tadpole responses to PPR. A positive correlation between behavioural and morphological responses in studies where both were measured indicates that trait co‐specialization is the predominant pattern of defense deployment in larval amphibians. This positive relationship suggests that survival tends to be maximized in tadpoles through equivalent co‐activation of multiple independent axes of protection, opposed to maximal expression along any single axis.

Synthesis

Our understanding of plastic responses to perceived predation risk (PPR) has benefited substantially from the vast amount of experimental work examining inducible defences in anuran tadpoles. Indeed this research has illustrated the wide variety of ways that prey animals can respond to the same risk event. We conducted a metaanalysis to identify the key ecological and experimental determinants of inducible defence expression. We then show that, in most cases, behavioural and morphological responses to PPR tend to be co‐expressed suggesting that responding along one axis (moving behaviour) does not limit their ability to respond along another distinct axis (tail morphology).
Optimal Design of Experiments on Connected Units with Application to Social Networks
Authors: Parker Ben M. , Gilmour Steven G. , Schormans John
Published: August 2016

When experiments are performed on social networks, it is difficult to justify the usual assumption of treatment–unit additivity, owing to the connections between actors in the network. We investigate how connections between experimental units affect the design of experiments on those experimental units. Specifically, where we have unstructured treatments, whose effects propagate according to a linear network effects model which we introduce, we show that optimal designs are no longer necessarily balanced; we further demonstrate how experiments which do not take a network effect into account can lead to much higher variance than necessary and/or a large bias. We show the use of this methodology in a very wide range of experiments in agricultural trials, and crossover trials, as well as experiments on connected individuals in a social network.
Macro‐scale stability with micro‐scale diversity: modelling changing ethnic minority residential segregation – London 2001–2011
Authors: Johnston Ron , Jones Kelvyn , Manley David , Owen Dewi
Published: August 2016

Most studies of ethnic residential segregation that address the issue of spatial scale make it implicit – if not explicit – that segregation is greater at smaller than larger scales. Such studies, however, invariably measure segregation separately at those scales, and take no account of the fact that measures at the smaller scale necessarily incorporate that at any larger scales. The present paper rectifies that situation by, for the first time, modelling ethnic segregation in London at the 2001 and 2011 censuses within a Bayesian statistical framework at three scales, which allows for the statistical significance of any changes to be formally assessed – something not possible before. It finds that for many of the groups studied segregation was as great, if not greater, at the macro‐scale as at the micro‐scale, with both measures larger than at the meso‐scale, with significant reductions in segregation across the decade, especially at the micro‐scale.
Hepatitis E virus infection in the Irish blood donor population
Authors: O'Riordan Joan , Boland Fiona , Williams Padraig , Donnellan Joe , Hogema Boris M. , Ijaz Samreen , Murphy William G.
Published: August 2016

BACKGROUND

Hepatitis E virus (HEV) Genotype 3 (G3) infection is a zoonosis that may be transmitted during the acute phase by transfusion. The aim of this study was to determine the incidence of HEV and seroprevalence among Irish blood donors.

STUDY DESIGN AND METHODS

Anonymized samples from 1076 donations collected in 2012 were tested for HEV immunoglobulin (Ig)G using the Wantai enzyme‐linked immunosorbent assay. A total of 24,985 anonymized donations collected between December 2013 and June 2014 were individually tested for HEV RNA using the Procleix HEV assay; reactive donations were confirmed by an in‐house real‐time polymerase chain reaction (PCR) test.

RESULTS

Seroprevalence for anti‐IgG was 5.3% (95% confidence interval [CI], 4.0%‐6.8%), ranging from 1.1% in the 18‐ to 29‐years age group to 33.3% in males over 60 years. HEV RNA screening of 24,985 samples yielded five PCR‐confirmed donations (1:4997, 0.02%; 95% CI, 0.0065%‐0.0467%), only one of which was serologically reactive (HEV IgM reactive only). Viral loads ranged from 10 to 44,550 IU/mL. Genotype analysis on three samples identified HEV G3 virus. Four of the five viremic donations were from donors in the 18‐ to 29‐years age group (p = 0.01).

CONCLUSION

Seroprevalence for anti‐HEV IgG was low compared to some European countries, but 1 in 5000 donations was viremic. Viremia was predominantly in younger Irish donors. After Department of Health approval the Irish Blood Transfusion Service implemented individual blood donation HEV RNA screening initially for a 3‐year period from January 2016.
Orbital reconstruction in the dog, cat, and horse
Authors: Wallin‐Håkansson Nils , Berggren Karin
Published: August 2016

Objective

To describe an adaptable method for reconstruction of the orbit following partial orbitectomy.

Animals studied

One horse, one cat, and four dogs.

Procedure(s)

Following partial orbitectomy for removal of bone and soft tissue affected by pathologic processes, reconstruction was achieved. Cerclage wires were used to reconstitute the orbital rim and other salient facial contours involved in excisions. These wires were then covered with a prolene mesh, first inside the orbit and then outwards over the affected extraorbital areas. Thereafter, a collagen sheet was placed over the mesh. Finally, subcutis and skin were closed over the construct.

Results

All operated eyes remained visual with normal position, direction, and mobility. Eyelid function, tear production, and nasolacrimal function were preserved. Side effects were mild and temporary, but animals requiring a lateral‐posterior surgical approach experienced concavity to the side of the head posterior to the orbital ligament region. One bone tumor out of three recurred.

Conclusions

The reconstruction method presented offers excellent results tectonically, cosmetically, and functionally, even following extensive orbitectomy. By adapted application of three reconstruction steps using readily available materials, large defects may be surgically repaired. Once orbitectomy is mastered, reconstruction requires no additional specialized techniques or equipment.
Ahmed glaucoma valve implantation with Ologen®Collagen Matrix for the surgical treatment of feline glaucoma
Authors: Park Kang‐hyo , Kim Joon‐Young , Choo Seoung‐woo , Yoon Hun‐young , Jeong Soon‐wuk
Published: August 2016

An Ahmed valve implantation with an Ologen®Collagen Matrix (Ologen®CM, Aeon Astron, Leiden, the Netherlands) was performed for the treatment of uncontrolled glaucoma in a cat. This cat was a 5‐year‐old castrated Russian Blue male with a 12‐week history of conjunctival hyperemia and mydriasis of the left eye. During the ophthalmic examination, the intraocular pressure (IOP) oculus sinister (OS) was 52 mmHg, and a narrow iridocorneal angle (ICA) was detected by gonioscopy. Medical treatment with Cosopt®(2% dorzolamide and 0.5% timolol) failed to decrease theIOP. The left eye still had vision, and an Ahmed valve implantation was performed. During the gonioimplantation, Ologen®CMwas used to inhibit scar formation around the valve. Following the operation, theIOPwas stable at an approximate average of 15 mmHg during the 7‐month follow‐up period, and vision in the left eye was retained without medication. An adequate subconjunctival filtering bleb was formed after 140 days. This is the first case report in which an Ahmed valve gonioimplant with an Ologen®CMhas been used for the surgical treatment of glaucoma in a cat.
White blood cell concentrations during lower body negative pressure and blood loss in humans
Authors: van Helmond Noud , Johnson Blair D. , Curry Timothy B. , Cap Andrew P. , Convertino Victor A. , Joyner Michael J.
Published: August 2016

New Findings

What is the central question of this study?

Is lower body negative pressure a useful surrogate to study white blood cell responses to haemorrhage in humans?

What is the main finding and its importance?

We found that lower body negative pressure appears to be a useful surrogate to study the early white blood cell mobilization response during blood loss.

Hypovolaemia has been associated with an immune response that might be secondary to sympathoexcitation. We tested the hypothesis that simulated hypovolaemia using lower body negative pressure (LBNP) and real hypovolaemia induced via experimental blood loss (BL) cause similar increases in the white blood cell concentration ([WBC]). We measured [WBC] and catecholamine concentrations in 12 men who underwent an LBNP and a BL protocol in a randomized order. We compared 45 mmHg of LBNP with 1000 ml of BL; therefore, [WBC] and catecholamine concentrations were plotted against central venous pressure to obtain stimulus–response relationships using the linear regression line slopes for both protocols. Mean regression line slopes were similar for total [WBC] (LBNP 183 ± 4 μl−1 mmHg−1 versus BL 155 ± 109 μl−1 mmHg−1, P = 0.15), neutrophils (LBNP 110 ± 2 μl−1 mmHg−1 versus BL 96 ± 72 μl−1 mmHg−1, P = 0.15) and lymphocytes (LBNP 65 ± 21 μl−1 mmHg−1 versus BL 59 ± 38 μl−1 mmHg−1, P = 0.90). Mean regression line slopes for adrenaline were similar (LBNP 15 ± 5 pg ml−1 mmHg−1 versus BL 16 ± 4 pg ml−1 mmHg−1, P = 0.84) and were steeper during LBNP for noradrenaline (LBNP 28 ± 6 pg ml−1 mmHg−1 versus BL 9 ± 6 pg ml−1 mmHg−1, P = 0.01). These data indicate that central hypovolaemia elicits a relative leucocytosis with a predominantly neutrophil‐based response. Additionally, our results indicate that LBNP models the stimulus–response relationship between central venous pressure and [WBC] observed during BL.
Learnability of prosodic boundaries: Is infant-directed speech easier?
Authors: Ludusan Bogdan , Cristia Alejandrina , Martin Andrew , Mazuka Reiko , Dupoux Emmanuel
Published: August 2016

This study explores the long-standing hypothesis that the acoustic cues to prosodic boundaries in infant-directed speech (IDS) make those boundaries easier to learn than those in adult-directed speech (ADS). Three cues (pause duration, nucleus duration, and pitch change) were investigated, by means of a systematic review of the literature, statistical analyses of a corpus of Japanese, and machine learning experiments. The review of previous work revealed that the effect of register on boundary cues is less well established than previously thought, and that results often vary across studies for certain cues. Statistical analyses run on a large database of mother-child and mother-interviewer interactions showed that the duration of a pause and the duration of the syllable nucleus preceding the boundary are two cues which are enhanced in IDS, while f0 change is actually degraded in IDS. Supervised and unsupervised machine learning techniques applied to these acoustic cues revealed that IDS boundaries were consistently better classified than ADS ones, regardless of the learning method used. The role of the cues examined in this study and the importance of these findings in the more general context of early linguistic structure acquisition is discussed.
Structural photoactivation of a full-length bacterial phytochrome
Authors: Björling Alexander , Berntsson Oskar , Lehtivuori Heli , Takala Heikki , Hughes Ashley J. , Panman Matthijs , Hoernke Maria , Niebling Stephan , Henry Léocadie , Henning Robert , Kosheleva Irina , Chukharev Vladimir , Tkachenko Nikolai V. , Menzel Andreas , Newby Gemma , Khakhulin Dmitry , Wulff Michael , Ihalainen Janne A. , Westenhoff Sebastian
Published: August 2016

Time-resolved x-ray solution scattering reveals the conformational signaling mechanism of a bacterial phytochrome.
Identification of the Main Regulator Responsible for Synthesis of the Typical Yellow Pigment Produced by Trichoderma reesei
Authors: Derntl Christian , Rassinger Alice , Srebotnik Ewald , Mach Robert L. , Mach-Aigner Astrid R.
Published: August 2016

The industrially used ascomycete
Trichoderma reesei
secretes a typical yellow pigment during cultivation, while other
Trichoderma
species do not. A comparative genomic analysis suggested that a putative secondary metabolism cluster, containing two polyketide-synthase encoding genes, is responsible for the yellow pigment synthesis. This cluster is conserved in a set of rather distantly related fungi, including
Acremonium chrysogenum
and
Penicillium chrysogenum
. In an attempt to silence the cluster in
T. reesei
, two genes of the cluster encoding transcription factors were individually deleted. For a complete genetic proof-of-function, the genes were reinserted into the genomes of the respective deletion strains. The deletion of the first transcription factor (termed yellow pigment regulator 1 [Ypr1]) resulted in the full abolishment of the yellow pigment formation and the expression of most genes of this cluster. A comparative high-pressure liquid chromatography (HPLC) analysis of supernatants of the
ypr1
deletion and its parent strain suggested the presence of several yellow compounds in
T. reesei
that are all derived from the same cluster. A subsequent gas chromatography/mass spectrometry analysis strongly indicated the presence of sorbicillin in the major HPLC peak. The presence of the second transcription factor, termed yellow pigment regulator 2 (Ypr2), reduces the yellow pigment formation and the expression of most cluster genes, including the gene encoding the activator Ypr1.

IMPORTANCE
Trichoderma reesei
is used for industry-scale production of carbohydrate-active enzymes. During growth, it secretes a typical yellow pigment. This is not favorable for industrial enzyme production because it makes the downstream process more complicated and thus increases operating costs. In this study, we demonstrate which regulators influence the synthesis of the yellow pigment. Based on these data, we also provide indication as to which genes are under the control of these regulators and are finally responsible for the biosynthesis of the yellow pigment. These genes are organized in a cluster that is also found in other industrially relevant fungi, such as the two antibiotic producers
Penicillium chrysogenum
and
Acremonium chrysogenum
. The targeted manipulation of a secondary metabolism cluster is an important option for any biotechnologically applied microorganism.
Understanding the Streptococcus mutans Cid/Lrg System through CidB Function
Authors: Ahn Sang-Joon , Rice Kelly C.
Published: August 2016

The
Streptococcus mutans
lrgAB
and
cidAB
operons have been previously described as a potential model system to dissect the complexity of biofilm development and virulence of
S. mutans
. Herein, we have attempted to further characterize the Cid/Lrg system by focusing on CidB, which has been shown to be critical for the ability of
S. mutans
to survive and persist in a nonpreferred oxygen-enriched condition. We have found that the expression level of
cidB
is critical to oxidative stress tolerance of
S. mutans
, most likely by impacting
lrg
expression. Intriguingly, the impaired aerobic growth phenotype of the
cidB
mutant could be restored by the additional loss of either CidA or LrgA. Growth-dependent expression of
cid
and
lrg
was demonstrated to be tightly under the control of both CcpA and the VicKR two-component system (TCS), regulators known to play an essential role in controlling major catabolic pathways and cell envelope homeostasis, respectively. RNA sequencing (RNA-Seq) analysis revealed that mutation of
cidB
resulted in global gene expression changes, comprising major domains of central metabolism and virulence processes, particularly in those involved with oxidative stress resistance. Loss of CidB also significantly changed the expression of genes related to genomic islands (GI) TnSmu1 and TnSmu2, the CRISPR (clustered regularly interspaced short palindromic repeats)-Cas system, and toxin-antitoxin (T/A) modules. Taken together, these data show that CidB impinges on the stress response, as well as the fundamental cellular physiology of
S. mutans
, and further suggest a potential link between Cid/Lrg-mediated cellular processes,
S. mutans
pathogenicity, and possible programmed growth arrest and cell death mechanisms.

IMPORTANCE
The ability of
Streptococcus mutans
to survive a variety of harmful or stressful conditions and to emerge as a numerically significant member of stable oral biofilm communities are essential elements for its persistence and cariogenicity. In this study, the homologous
cidAB
and
lrgAB
operons, previously identified as being highly balanced and coordinated during
S. mutans
aerobic growth, were further characterized through the functional and transcriptomic analysis of CidB. Precise control of CidB levels is shown to impact the expression of
lrg
, oxidative stress tolerance, major metabolic domains, and the molecular modules linked to cell death and lysis. This study advances our understanding of the Cid/Lrg system as a key player in the integration of complex environmental signals (such as oxidative stress) into the regulatory networks that modulate
S. mutans
virulence and cell homeostasis.
Core Genome Multilocus Sequence Typing for Identification of Globally Distributed Clonal Groups and Differentiation of Outbreak Strains of Listeria monocytogenes
Authors: Chen Yi , Gonzalez-Escalona Narjol , Hammack Thomas S. , Allard Marc W. , Strain Errol A. , Brown Eric W.
Published: August 2016

Many listeriosis outbreaks are caused by a few globally distributed clonal groups, designated clonal complexes or epidemic clones, of
Listeria monocytogenes
, several of which have been defined by classic multilocus sequence typing (MLST) schemes targeting 6 to 8 housekeeping or virulence genes. We have developed and evaluated core genome MLST (cgMLST) schemes and applied them to isolates from multiple clonal groups, including those associated with 39 listeriosis outbreaks. The cgMLST clusters were congruent with MLST-defined clonal groups, which had various degrees of diversity at the whole-genome level. Notably, cgMLST could distinguish among outbreak strains and epidemiologically unrelated strains of the same clonal group, which could not be achieved using classic MLST schemes. The precise selection of cgMLST gene targets may not be critical for the general identification of clonal groups and outbreak strains. cgMLST analyses further identified outbreak strains, including those associated with recent outbreaks linked to contaminated French-style cheese, Hispanic-style cheese, stone fruit, caramel apple, ice cream, and packaged leafy green salad, as belonging to major clonal groups. We further developed lineage-specific cgMLST schemes, which can include accessory genes when core genomes do not possess sufficient diversity, and this provided additional resolution over species-specific cgMLST. Analyses of isolates from different common-source listeriosis outbreaks revealed various degrees of diversity, indicating that the numbers of allelic differences should always be combined with cgMLST clustering and epidemiological evidence to define a listeriosis outbreak.

IMPORTANCE
Classic multilocus sequence typing (MLST) schemes targeting internal fragments of 6 to 8 genes that define clonal complexes or epidemic clones have been widely employed to study
L. monocytogenes
biodiversity and its relation to pathogenicity potential and epidemiology. We demonstrated that core genome MLST schemes can be used for the simultaneous identification of clonal groups and the differentiation of individual outbreak strains and epidemiologically unrelated strains of the same clonal group. We further developed lineage-specific cgMLST schemes that targeted more genomic regions than the species-specific cgMLST schemes. Our data revealed the genome-level diversity of clonal groups defined by classic MLST schemes. Our identification of U.S. and international outbreaks caused by major clonal groups can contribute to further understanding of the global epidemiology of
L. monocytogenes
.
Heterologous Production of Cyanobacterial Mycosporine-Like Amino Acids Mycosporine-Ornithine and Mycosporine-Lysine in Escherichia coli
Authors: Katoch Meenu , Mazmouz Rabia , Chau Rocky , Pearson Leanne A. , Pickford Russell , Neilan Brett A.
Published: August 2016

Mycosporine-like amino acids (MAAs) are an important class of secondary metabolites known for their protection against UV radiation and other stress factors. Cyanobacteria produce a variety of MAAs, including shinorine, the active ingredient in many sunscreen creams. Bioinformatic analysis of the genome of the soil-dwelling cyanobacterium
Cylindrospermum stagnale
PCC 7417 revealed a new gene cluster with homology to MAA synthase from
Nostoc punctiforme
. This newly identified gene cluster is unusual because it has five biosynthesis genes (
mylA
to
mylE
), compared to the four found in other MAA gene clusters. Heterologous expression of
mylA
to
mylE
in
Escherichia coli
resulted in the production of mycosporine-lysine and the novel compound mycosporine-ornithine. To our knowledge, this is the first time these compounds have been heterologously produced in
E. coli
and structurally characterized via direct spectral guidance. This study offers insight into the diversity, biosynthesis, and structure of cyanobacterial MAAs and highlights their amenability to heterologous production methods.

IMPORTANCE
Mycosporine-like amino acids (MAAs) are significant from an environmental microbiological perspective as they offer microbes protection against a variety of stress factors, including UV radiation. The heterologous expression of MAAs in
E. coli
is also significant from a biotechnological perspective as MAAs are the active ingredient in next-generation sunscreens.
Effect of Leaf Surface Chemical Properties on Efficacy of Sanitizer for Rotavirus Inactivation
Authors: Fuzawa Miyu , Ku Kang-Mo , Palma-Salgado Sindy Paola , Nagasaka Kenya , Feng Hao , Juvik John A. , Sano Daisuke , Shisler Joanna L. , Nguyen Thanh H.
Published: August 2016

The use of sanitizers is essential for produce safety. However, little is known about how sanitizer efficacy varies with respect to the chemical surface properties of produce. To answer this question, the disinfection efficacies of an oxidant-based sanitizer and a new surfactant-based sanitizer for porcine rotavirus (PRV) strain OSU were examined. PRV was attached to the leaf surfaces of two kale cultivars with high epicuticular wax contents and one cultivar of endive with a low epicuticular wax content and then treated with each sanitizer. The efficacy of the oxidant-based sanitizer correlated with leaf wax content as evidenced by the 1-log
10
PRV disinfection on endive surfaces (low wax content) and 3-log
10
disinfection of the cultivars with higher wax contents. In contrast, the surfactant-based sanitizer showed similar PRV disinfection efficacies (up to 3 log
10
) that were independent of leaf wax content. A statistical difference was observed with the disinfection efficacies of the oxidant-based sanitizer for suspended and attached PRV, while the surfactant-based sanitizer showed similar PRV disinfection efficacies. Significant reductions in the entry and replication of PRV were observed after treatment with either disinfectant. Moreover, the oxidant-based-sanitizer-treated PRV showed sialic acid-specific binding to the host cells, whereas the surfactant-based sanitizer increased the nonspecific binding of PRV to the host cells. These findings suggest that the surface properties of fresh produce may affect the efficacy of virus disinfection, implying that food sanitizers should be carefully selected for the different surface characteristics of fresh produce.

IMPORTANCE
Food sanitizer efficacies are affected by the surface properties of vegetables. This study evaluated the disinfection efficacies of two food sanitizers, an oxidant-based sanitizer and a surfactant-based sanitizer, on porcine rotavirus strain OSU adhering to the leaf epicuticular surfaces of high- and low-wax-content cultivars. The disinfection efficacy of the oxidant-based sanitizer was affected by the surface properties of the vegetables, while the surfactant-based sanitizer was effective for both high- and low-wax leafy vegetable cultivars. This study suggests that the surface properties of vegetables may be an important factor that interacts with disinfection with food sanitizers of rotaviruses adhering to fresh produce.
Copper Resistance of the Emerging Pathogen Acinetobacter baumannii
Authors: Williams Caitlin L. , Neu Heather M. , Gilbreath Jeremy J. , Michel Sarah L. J. , Zurawski Daniel V. , Merrell D. Scott
Published: August 2016

Acinetobacter baumannii
is an important emerging pathogen that is capable of causing many types of severe infection, especially in immunocompromised hosts. Since
A. baumannii
can rapidly acquire antibiotic resistance genes, many infections are on the verge of being untreatable, and novel therapies are desperately needed. To investigate the potential utility of copper-based antibacterial strategies against
Acinetobacter
infections, we characterized copper resistance in a panel of recent clinical
A. baumannii
isolates. Exposure to increasing concentrations of copper in liquid culture and on solid surfaces resulted in dose-dependent and strain-dependent effects; levels of copper resistance varied broadly across isolates, possibly resulting from identified genotypic variation among strains. Examination of the growth-phase-dependent effect of copper on
A. baumannii
revealed that resistance to copper increased dramatically in stationary phase. Moreover,
A. baumannii
biofilms were more resistant to copper than planktonic cells but were still susceptible to copper toxicity. Exposure of bacteria to subinhibitory concentrations of copper allowed them to better adapt to and grow in high concentrations of copper; this copper tolerance response is likely achieved via increased expression of copper resistance mechanisms. Indeed, genomic analysis revealed numerous putative copper resistance proteins that share amino acid homology to known proteins in
Escherichia coli
and
Pseudomonas aeruginosa
. Transcriptional analysis revealed significant upregulation of these putative copper resistance genes following brief copper exposure. Future characterization of copper resistance mechanisms may aid in the search for novel antibiotics against
Acinetobacter
and other highly antibiotic-resistant pathogens.

IMPORTANCE
Acinetobacter baumannii
causes many types of severe nosocomial infections; unfortunately, some isolates have acquired resistance to almost every available antibiotic, and treatment options are incredibly limited. Copper is an essential nutrient but becomes toxic at high concentrations. The inherent antimicrobial properties of copper give it potential for use in novel therapeutics against drug-resistant pathogens. We show that
A. baumannii
clinical isolates are sensitive to copper
in vitro
, both in liquid and on solid metal surfaces. Since bacterial resistance to copper is mediated though mechanisms of efflux and detoxification, we identified genes encoding putative copper-related proteins in
A. baumannii
and showed that expression of some of these genes is regulated by the copper concentration. We propose that the antimicrobial effects of copper may be beneficial in the development of future therapeutics that target multidrug-resistant bacteria.
Minor Coat and Heat Shock Proteins Are Involved in the Binding of Citrus Tristeza Virus to the Foregut of Its Aphid Vector, Toxoptera citricida
Authors: Killiny N. , Harper S. J. , Alfaress S. , El Mohtar C. , Dawson W. O.
Published: August 2016

Vector transmission is a critical stage in the viral life cycle, yet for most plant viruses how they interact with their vector is unknown or is explained by analogy with previously described relatives. Here we examined the mechanism underlying the transmission of citrus tristeza virus (CTV) by its aphid vector,
Toxoptera citricida
, with the objective of identifying what virus-encoded proteins it uses to interact with the vector. Using fluorescently labeled virions, we demonstrated that CTV binds specifically to the lining of the cibarium of the aphid. Through
in vitro
competitive binding assays between fluorescent virions and free viral proteins, we determined that the minor coat protein is involved in vector interaction. We also found that the presence of two heat shock-like proteins, p61 and p65, reduces virion binding
in vitro
. Additionally, treating the dissected mouthparts with proteases did not affect the binding of CTV virions. In contrast, chitinase treatment reduced CTV binding to the foregut. Finally, competition with glucose,
N
-acetyl-β-
d
-glucosamine, chitobiose, and chitotriose reduced the binding. These findings together suggest that CTV binds to the sugar moieties of the cuticular surface of the aphid cibarium, and the binding involves the concerted activity of three virus-encoded proteins.

IMPORTANCE
Limited information is known about the specific interactions between citrus tristeza virus and its aphid vectors. These interactions are important for the process of successful transmission. In this study, we localized the CTV retention site as the cibarium of the aphid foregut. Moreover, we demonstrated that the nature of these interactions is protein-carbohydrate binding. The viral proteins, including the minor coat protein and two heat shock proteins, bind to sugar moieties on the surface of the foregut. These findings will help in understanding the transmission mechanism of CTV by the aphid vector and may help in developing control strategies which interfere with the CTV binding to its insect vector to block the transmission.
Molecular Mechanisms Contributing to the Growth and Physiology of an Extremophile Cultured with Dielectric Heating
Authors: Cusick Kathleen D. , Lin Baochuan , Malanoski Anthony P. , Strycharz-Glaven Sarah M. , Cockrell-Zugell Allison , Fitzgerald Lisa A. , Cramer Jeffrey A. , Barlow Daniel E. , Boyd Thomas J. , Biffinger Justin C.
Published: August 2016

The effect of microwave frequency electromagnetic fields on living microorganisms is an active and highly contested area of research. One of the major drawbacks to using mesophilic organisms to study microwave radiation effects is the unavoidable heating of the organism, which has limited the scale (<5 ml) and duration (<1 h) of experiments. However, the negative effects of heating a mesophile can be mitigated by employing thermophiles (organisms able to grow at temperatures of >60°C). This study identified changes in global gene expression profiles during the growth ofThermus scotoductusSA-01 at 65°C using dielectric (2.45 GHz, i.e., microwave) heating. RNA sequencing was performed on cultures at 8, 14, and 24 h after inoculation to determine the molecular mechanisms contributing to long-term cellular growth and survival under microwave heating conditions. Over the course of growth, genes associated with amino acid metabolism, carbohydrate metabolism, and defense mechanisms were upregulated; the number of repressed genes with unknown function increased; and at all time points, transposases were upregulated. Genes involved in cell wall biogenesis and elongation were also upregulated, consistent with the distinct elongated cell morphology observed after 24 h using microwave heating. Analysis of the global differential gene expression data enabled the identification of molecular processes specific to the response ofT. scotoductusSA-01 to dielectric heating during growth.

IMPORTANCEThe residual heating of living organisms in the microwave region of the electromagnetic spectrum has complicated the identification of radiation-only effects using microorganisms for 50 years. A majority of the previous experiments used either mature cells or short exposure times with low-energy high-frequency radiation. Using global differential gene expression data, we identified molecular processes unique to dielectric heating usingThermus scotoductusSA-01 cultured over 30 h in a commercial microwave digestor. Genes associated with amino acid metabolism, carbohydrate metabolism, and defense mechanisms were upregulated; the number of repressed genes with unknown function increased; and at all time points, transposases were upregulated. These findings serve as a platform for future studies with mesophiles in order to better understand the response of microorganisms to microwave radiation.
Peptidoglycan Acetylation of Campylobacter jejuni Is Essential for Maintaining Cell Wall Integrity and Colonization in Chicken Intestines
Authors: Iwata Taketoshi , Watanabe Ayako , Kusumoto Masahiro , Akiba Masato
Published: August 2016

Peptidoglycan (PG) acetylation of Gram-positive bacteria confers lysozyme resistance and contributes to survival in the host. However, the importance of PG acetylation in Gram-negative bacteria has not been fully elucidated. The genes encoding putative PG acetyltransferase A (PatA) and B (PatB) are highly conserved in
Campylobacter jejuni
, the predominant cause of bacterial diarrhea worldwide. To evaluate the importance of PatA and PatB of
C. jejuni
, we constructed
patA
and
patB
isogenic mutants and compared their phenotypes with those of the parental strains. Although transmission electron microscopy did not reveal morphological changes, both mutants exhibited decreased motility and biofilm formation
in vitro
. The extent of acetylation of the PG purified from the
patA
and
patB
mutants was significantly lower than the PG acetylation in the parental strains. Both mutants exhibited decreased lysozyme resistance and intracellular survival in macrophage cells. In a chick colonization experiment, significant colonization deficiency was observed for both mutants. These results suggest that PatA and PatB of
C. jejuni
play important roles in maintaining cell wall integrity by catalyzing PG
O
-acetylation and that the loss of these enzymes causes decreased motility and biofilm formation, thus leading to colonization deficiency in chicken infection.

IMPORTANCE
The importance of peptidoglycan (PG) acetylation in Gram-negative bacteria has not been fully elucidated. The genes encoding putative PG acetyltransferase A (PatA) and B (PatB) are highly conserved in
Campylobacter jejuni
, the predominant cause of bacterial diarrhea worldwide. We evaluated the importance of these enzymes using isogenic mutants. The results of this study suggest that PatA and PatB of
C. jejuni
play important roles in maintaining cell wall integrity. The loss of these factors caused multiple phenotypic changes, leading to colonization deficiency in chicken infection. These data should be useful in developing novel control measures to prevent chicken colonization by
C. jejuni
. Inhibitors of the PG acetylation enzymes PatA and PatB might serve as potent anti-
C. jejuni
agents.
Introducing Vibro-Acupuncture: A Psychophysical Study
Authors: Wang Kelun , Larsen Dennis Boye , Jensen Tanja Kim , Liu Tiejun , Liu Zhuxuan , Arendt-Nielsen Lars
Published: August 2016

Background/aim

To potentially enhance the effects of conventional acupuncture, a novel acu-vibrator (prototype) has been developed to perform vibro-acupuncture (VA). The aim of this psychophysical study was to investigate the subjective sensations of VA compared with conventional manual acupuncture (MA) and non-penetrating sham acupuncture (SA).

Methods

30 young healthy volunteers (21 men and 9 women) received VA, MA, and SA at LI4 and LI10 in a randomised, single-blind, placebo-controlled, cross-over manner. After 25 min of treatment, the Massachusetts General Hospital (MGH) acupuncture sensation scale (MASS), McGill pain questionnaire (MPQ), and numerical rating scale (NRS; 0–10) were employed followed by the acupuncture credibility and indication scale. Adverse events were investigated after treatment. Data were analysed using Friedman's test for repeated measures on ranks and post hoc Wilcoxon signed-rank tests with Bonferroni correction.

Results

The MASS scores were significantly higher during MA and VA compared with SA at both LI4 and LI10 (p<0.017). Treatment with VA evoked significantly higher vibration sensations compared with MA and SA (p<0.005). Treatment with SA yielded significantly lower NRS and MPQ scores compared with MA and VA (p<0.001) with no difference between MA and VA (p>0.05). Blinding of participants was achieved for SA and MA; however, VA was correctly identified in 29 of 30 subjects due to the characteristic vibrational stimulation. No serious adverse events were recorded for any of the treatments.

Conclusions

Subjective sensations were influenced by treatment mode, with MA and VA yielding higher stimulation responses compared with SA. VA evoked specific vibrational sensations beyond MA, which might have specific effects in various disorders.