bluuebunny/crossref_metadata_2025 huggingface dataset:
Pharmacokinetics of posaconazole in koalas (Phascolarctos cinereus) after intravenous and oral administration
Authors: Gharibi S. , Kimble B. , Vogelnest L. , Barnes J. , Stadler C. K. , Govendir M.
Published: April 2017

The pharmacokinetic profile of posaconazole in clinically normal koalas (n = 8) was investigated. Single doses of posaconazole were administered intravenously (i.v.; 3 mg/kg; n = 2) or orally (p.o.; 6 mg/kg; n = 6) with serial plasma samples collected over 24 and 36 hr, respectively. Plasma concentrations of posaconazole were quantified by validated high‐performance liquid chromatography. A noncompartmental pharmacokinetic analysis of data was performed. Following i.v. administration, estimates of the median (range) of plasma clearance (CL) and steady‐state volume of distribution (Vss) were 0.15 (0.13–0.18) L hr−1 kg−1 and 1.23 (0.93–1.53) L/kg, respectively. The median (range) elimination half‐life (t1/2) after i.v. and p.o. administration was 7.90 (7.62–8.18) and 12.79 (11.22–16.24) hr, respectively. Oral bioavailability varied from 0.43 to 0.99 (median: 0.66). Following oral administration, maximum plasma concentration (Cmax; median: 0.72, range: 0.55–0.93 μg/ml) was achieved in 8 (range 6–12) hr. The in vitro plasma protein binding of posaconazole incubated at 37°C was 99.25 ± 0.29%. Consideration of posaconazole pharmacokinetic/pharmacodynamic (PK/PD) targets for some yeasts such as disseminated candidiasis suggests that posaconazole could be an efficacious treatment for cryptococcosis in koalas.
Emergency department utilization in pediatric heart transplant recipients
Authors: Wittlieb‐Weber Carol A. , Rossano Joseph W. , Weber David R. , Lin Kimberly Y. , Ravishankar Chitra , Mascio Christopher E. , Shaddy Robert E. , O'Connor Matthew J.
Published: April 2017

We used the NEDS database (2010) to evaluate ED utilization in PED HT recipients compared to other patient populations with focus on characteristics of ED visits, risk factors for admission, and charges. We analyzed 433 ED visits by PED HT recipients (median age 8 [range: 0‐18] years). The most common primary diagnosis category was infectious (n=163, 37.6%), with pneumonia being the most common infectious etiology. When compared to all PED visits, HT visits were more likely to result in hospital admission (32.6% versus 3.9%, P<.001), had greater hospital LOS (median of 3 days [IQR 2‐4] versus 2 days [IQR 1‐4], P=.001), and accumulated greater total hospital charges (median $26 317 [IQR $11 438‐$46 407] versus $12 332 [IQR $7092‐$22 583], P<.001). When compared to visits by other SOT recipients, results varied with similar rates of hospital admission for HT, LUNGT, and KT visits and similar LOS for HT and KT visits but differing total hospital charges. Although PED HT recipients account for a small percentage of overall ED visits, they are more likely to be hospitalized and require greater resource utilization compared to the general PED population, but not when compared to other SOT recipients.
High‐milking sheep have a lower ovulation rate and tend to yield fewer embryos in response to superovulation and intrauterine artificial insemination
Authors: Meraï A , Dattena M , Casu S , Rekik M , Lassoued N
Published: April 2017

Contents

Antagonistic relationship between milk yield and reproduction is reported in several livestock species. This study aimed to investigate whether genetic merit for milk production in dairy sheep affects responses to superovulation, embryo yield and quality. A total of 21 cross‐bred Sarda x Lacaune ewes homogeneous for age, parity and stage of lactation were included. The ewes were stratified as high‐producing or low‐producing based on their genetic merit for milk production estimated by a pentatrait repeatability animal model. Oestrus was synchronized using an intravaginal progesterone pessary inserted on Day 0 and removed on Day 14. Superovulatory treatment consisted of 350 I.U. of porcine FSH administered in eight decreasing intramuscular doses every 12 hr with a total dose of 10 ml of solution starting 12 days after insertion of sponges. Laparoscopic artificial insemination (AI) was performed 48 hr after pessary removal. Surgical embryo recovery was performed at Day 8 after pessary removal. Correlation between breeding value for milk production and the number of corpora lutea (CL) was significantly different from zero (−0.49). High‐producing ewes had a lower number of CL than low‐producing counterparts (7.6 ± 2.50 vs 12.1 ± 5.16 respectively; p < .02). Furthermore, there was a tendency for high‐producing ewes to yield fewer embryos than low‐producing females (5.3 ± 3.46 vs 9.18 ± 5.11; p = .09). No differences were observed between ewes in both genetic groups with regard to the number of embryos of grades 1, 2 and 3. To our knowledge, this is the first report highlighting an antagonism between genetic merit for milk production and the ability to produce embryos in sheep. These results deserve to be considered in sheep breeding programmes.
Detection of intracellular reactive oxygen species (superoxide anion and hydrogen peroxide) and lipid peroxidation during cryopreservation of alpaca spermatozoa
Authors: Evangelista‐Vargas S , Santiani A
Published: April 2017

Contents

The objective of this study was to detect changes in intracellular reactive oxygen species (superoxide anion and hydrogen peroxide) production and lipid peroxidation during cryopreservation of alpaca spermatozoa. Twelve alpaca semen samples were conventionally cryopreserved. Intracellular superoxide anion and hydrogen peroxide were evaluated by fluorescence microscopy using dihydroethidium (DHE)/YO‐PRO‐1 and dichlorofluorescein diacetate (H2DCFDA)/propidium iodide (PI), respectively. Evaluations were performed during cooling curve at (1) 25°C, (2) 15°C, (3) 5°C/0 min, (4) 5°C/15 min, (5) 5°C/30 min and (6) after freezing/thawing. Evaluation of lipid peroxidation by measuring malondialdehyde (MDA) was performed at 25°C, 5°C/30 min and after thawing. Maximum percentages of total spermatozoa producing superoxide anion and hydrogen peroxide were found at 5°C/30 min (62.8 ± 6.3% and 30.5 ± 5.6%, respectively), and these results were higher (p < .05) than initial (25°C: 10.8 ± 3.8% and 6.8 ± 0.7%, respectively) and after thawing (29.8 ± 9.5% and 7.5 ± 1.8%, respectively) values. However, considering only viable spermatozoa, production of superoxide anion and hydrogen peroxide during overall stabilization at 5°C (>76% and >91%, respectively) and after thawing (74.9 ± 5.0% and 78.9 ± 2.2%, respectively) was higher (p < .05) than initial values at 25°C (38.7 ± 3.1% and 53.6 ± 2.0%, respectively). Lipid peroxidation at 25°C, 5°C/30 min, and post‐thawing were 346.5 ± 99.8, 401.1 ± 64.8 and 527.7 ± 142.8 ng/ml MDA, respectively. These results showed that high percentage of viable alpaca spermatozoa produces intracellular reactive species oxygen (ROS) during the cryopreservation process of alpaca semen.
Extended day length in late winter/early spring, with a return to natural day length of shorter duration, increased plasma testosterone and sexual performance in rams with or without melatonin implants
Authors: Abecia JA , Chemineau P , Keller M , Delgadillo JA
Published: April 2017

Contents

Sixteen rams were used to quantify the effects of long days, imposed during late winter/early spring, with or without exogenous melatonin, on plasma testosterone concentrations and ram serving capacity. Rams were assigned to two groups: photoperiod‐treated rams (Artificial Photoperiod, AP; n = 8), exposed to 2 months of long days (16 hr of light/day) between 22 December and 22 February, and control rams (Natural Photoperiod, NP; n = 8). At the end of the long‐day period, AP rams were returned to the natural photoperiod, and each ram in the two groups either did (+M) or did not (‐M) receive three subcutaneous melatonin implants. Four groups were created as follows: AP+M (n = 4), AP‐M (n = 4), NP+M (n = 4) and NP‐M (n = 4). Thirty days after of the onset of photoperiodic treatment, AP rams (13.5 ± 2.8 ng/ml) had significantly (p < .05) lower testosterone levels than NP rams (36.7 ± 1.0), and similar differences were not apparent at the end of the photoperiod treatment. A month later, AP rams (24.3 ± 7.9) had higher (p < .10) testosterone levels than NP rams (13.1 ± 5.0), with no effect of melatonin treatment. Fifty days after melatonin implantations, rams were exposed for 20 min to three oestrous ewes. AP rams (2.50 ± 0.42) exhibited significantly (p < .05) more serves than did NP rams (1.11 ± 0.39), and melatonin treatment had no significant effect; however, the interaction between treatments was significant. Time to first serve was significantly (p < .05) shorter in AP (2.30 ± 1.20 min) than it was in NP rams (5.58 ± 0.68 min). In conclusion, exposure to 2 months of long days in late winter/early spring, with a return to natural day length of shorter duration, increased plasma testosterone concentrations and sexual performance in rams with or without exogenous melatonin. This particular management is an option if a non‐hormonal reproductive strategy is scheduled; yet, if the use of exogenous hormones is feasible, melatonin implants increase the mating efficiency of rams.
Relationship between the risk factors for human papillomavirus infection and lower genital tract precursor lesion and cancer development in female transplant recipients
Authors: de Oliveira Martins Caroline Alves , Do Val Guimarães Isabel Cristina Chulvis , Velarde Luis Guillermo Coca
Published: April 2017

Background

This study aimed to analyze the relationship between the several risk factors for human papillomavirus (HPV) infection and its persistence and to assess the prevalence of the lower genital tract precursor lesions, cancer, and HPV infection in female transplant recipients, besides the most prevalent HPV types.

Methods

The methodology adopted was a cross‐sectional study with a random sample of 61 patients.

Results

The results indicated 10 cases (16.4%) of lesions, 54.5% of the overall prevalence of HPV infection, and HPV 16 was the most common high‐risk HPV type, followed by HPV 51/53/70. A multiple logistic regression was done and hormone use presented a statistically significant association with high‐risk HPV infection (P=.037). No statistically significant association was identified for the set of all factors with the lesions studied.

Conclusions

The high prevalence of HPV infection and its precursor lesions confirmed the importance of differential screening and follow‐up of transplanted patients. Condom use must be encouraged. The study result also confirmed the importance of knowledge of the viral type, in order to analyze the impact of HPV immunization, particularly against HPV types that are not 16/18. The reasonable rate of other genotypes, besides the vaccine types, should be considered to evaluate vaccination strategies.
Comparison of the rates of Clostridium difficile and bacteremia after delaying fluoroquinolone prophylaxis from day 0 to day +3 post autologous stem cell transplantation
Authors: Koseck Karin , Steinberg Amir , Caliendo Gina , Meyer Joanne , Kim Sara S.
Published: April 2017

Prophylactic fluoroquinolones are routinely administered after stem cell transplantation (SCT) to prevent bacterial infection; however, fluoroquinolones may increase the risk of Clostridium difficile infection, particularly in immunocompromised patients. This study is designed to evaluate the effect of a delay by 3 days in fluoroquinolone prophylaxis after autologous SCT (ASCT) on the rates of C. difficile infection and bacteremia. A single‐center retrospective cohort study was performed in 118 patients who received levofloxacin prophylaxis following ASCT at our institution between November 2014 and October 2015. In efforts to reduce the rate of C. difficile, initiation of levofloxacin prophylaxis was delayed from day 0 to day +3 of SCT beginning April 30, 2015. The incidence of C. difficile infection and of bacteremia in patients who initiated levofloxacin on day 0 was compared with those who started prophylaxis on day +3. We found no difference in the rates of C. difficile (7.9% vs 5.5%, P=.593) and bacteremia (7.9% vs 3.6%, P=.323) in patients who initiated levofloxacin on day 0 compared with those who initiated prophylaxis on day +3. Delaying the initiation of levofloxacin prophylaxis by 3 days post ASCT showed no difference in the incidence of C. difficile or bacteremia. Future studies are warranted to show feasibility of delaying the initiation of antibiotic prophylaxis until neutropenia post ASCT, to further minimize the duration of antibiotic exposure.
Does the intercept of the heat–stress relation provide an accurate estimate of cardiac activation heat?
Authors: Pham Toan , Tran Kenneth , Mellor Kimberley M , Hickey Anthony , Power Amelia , Ward Marie‐Louise , Taberner Andrew , Han June‐Chiew , Loiselle Denis
Published: April 2017

Key points

The heat of activation of cardiac muscle reflects the metabolic cost of restoring ionic homeostasis following a contraction. The accuracy of its measurement depends critically on the abolition of crossbridge cycling.

We abolished crossbridge activity in isolated rat ventricular trabeculae by use of blebbistatin, an agent that selectively inhibits myosin II ATPase.

We found cardiac activation heat to be muscle length independent and to account for 15–20% of total heat production at body temperature.

We conclude that it can be accurately estimated at minimal muscle length.

Activation heat arises from two sources during the contraction of striated muscle. It reflects the metabolic expenditure associated with Ca2+ pumping by the sarcoplasmic reticular Ca2+‐ATPase and Ca2+ translocation by the Na+/Ca2+ exchanger coupled to the Na+,K+‐ATPase. In cardiac preparations, investigators are constrained in estimating its magnitude by reducing muscle length to the point where macroscopic twitch force vanishes. But this experimental protocol has been criticised since, at zero force, the observed heat may be contaminated by residual crossbridge cycling activity. To eliminate this concern, the putative thermal contribution from crossbridge cycling activity must be abolished, at least at minimal muscle length. We achieved this using blebbistatin, a selective inhibitor of myosin II ATPase. Using a microcalorimeter, we measured the force production and heat output, as functions of muscle length, of isolated rat trabeculae from both ventricles contracting isometrically at 5 Hz and at 37°C. In the presence of blebbistatin (15 μmol l−1), active force was zero but heat output remained constant, at all muscle lengths. Activation heat measured in the presence of blebbistatin was not different from that estimated from the intercept of the heat–stress relation in its absence. We thus reached two conclusions. First, activation heat is independent of muscle length. Second, residual crossbridge heat is negligible at zero active force; hence, the intercept of the cardiac heat–force relation provides an estimate of activation heat uncontaminated by crossbridge cycling. Both results resolve long‐standing disputes in the literature.
Origami by frontal photopolymerization
Authors: Zhao Zeang , Wu Jiangtao , Mu Xiaoming , Chen Haosen , Qi H. Jerry , Fang Daining
Published: April 2017

A novel method to create 3D origami structures was developed using frontal photopolymerization with grayscale light.
Mitochondrial protein-linked DNA breaks perturb mitochondrial gene transcription and trigger free radical–induced DNA damage
Authors: Chiang Shih-Chieh , Meagher Martin , Kassouf Nick , Hafezparast Majid , McKinnon Peter J. , Haywood Rachel , El-Khamisy Sherif F.
Published: April 2017

Mitochondrial protein-linked DNA repair promotes gene transcription and protects from free radical–induced DNA damage.
Carbon-free H
2
production from ammonia triggered at room temperature with an acidic RuO
2
/γ-Al
2
O
3
catalyst
Authors: Nagaoka Katsutoshi , Eboshi Takaaki , Takeishi Yuma , Tasaki Ryo , Honda Kyoko , Imamura Kazuya , Sato Katsutoshi
Published: April 2017

Heat evolved by ammonia adsorption on RuO
2
/γ-Al
2
O
3
heats the catalyst from room temperature to the autoignition temperature.
Genuine binding energy of the hydrated electron
Authors: Luckhaus David , Yamamoto Yo-ichi , Suzuki Toshinori , Signorell Ruth
Published: April 2017

Water microjet photoemission experiments and scattering calculations reveal genuine properties of the hydrated electron.
A Novel Quorum-Quenching
N
-Acylhomoserine Lactone Acylase from Acidovorax sp. Strain MR-S7 Mediates Antibiotic Resistance
Authors: Kusada Hiroyuki , Tamaki Hideyuki , Kamagata Yoichi , Hanada Satoshi , Kimura Nobutada
Published: April 2017

N
-Acylhomoserine lactone acylase (AHL acylase) is a well-known enzyme responsible for disrupting cell-cell communication (quorum sensing) in bacteria. Here, we isolated and characterized a novel and unique AHL acylase (designated MacQ) from a multidrug-resistant bacterium,
Acidovorax
sp. strain MR-S7. The purified MacQ protein heterologously expressed in
Escherichia coli
degraded a wide variety of AHLs, ranging from C
6
to C
14
side chains with or without 3-oxo substitutions. We also observed that AHL-mediated virulence factor production in a plant pathogen,
Pectobacterium carotovorum
, was dramatically attenuated by coculture with MacQ-overexpressing
Escherichia coli
, whereas
E. coli
with an empty vector was unable to quench the pathogenicity, which strongly indicates that MacQ can act
in vivo
as a quorum-quenching enzyme and interfere with the quorum-sensing system in the pathogen. In addition, this enzyme was found to be capable of degrading a wide spectrum of β-lactams (penicillin G, ampicillin, amoxicillin, carbenicillin, cephalexin, and cefadroxil) by deacylation, clearly indicating that MacQ is a bifunctional enzyme that confers both quorum quenching and antibiotic resistance on strain MR-S7. MacQ has relatively low amino acid sequence identity to any of the known acylases (<39%) and has among the broadest substrate range. Our findings provide the possibility that AHL acylase genes can be an alternative source of antibiotic resistance genes posing a threat to human health if they migrate and transfer to pathogenic bacteria.

IMPORTANCE
N
-Acylhomoserine lactones (AHLs) are well-known signal molecules for bacterial cell-cell communication (quorum sensing), and AHL acylase, which is able to degrade AHLs, has been recognized as a major target for quorum-sensing interference (quorum quenching) in pathogens. In this work, we succeeded in isolating a novel AHL acylase (MacQ) from a multidrug-resistant bacterium and demonstrated that the MacQ enzyme could confer multidrug resistance as well as quorum quenching on the host organism. Indeed, the purified MacQ protein was found to be bifunctional and capable of degrading not only various AHL derivatives but also multiple β-lactam antibiotics by deacylation activities. Although quorum quenching and antibiotic resistance have been recognized to be distinct biological functions, our findings clearly link the two functions by discovering the novel bifunctional enzyme and further providing the possibility that a hitherto-overlooked antibiotic resistance mechanism mediated by the quorum-quenching enzyme may exist in natural environments and perhaps in clinical settings.
Novel Method for Reliable Identification of Siccibacter and Franconibacter Strains: from “Pseudo-Cronobacter” to New Enterobacteriaceae Genera
Authors: Svobodová Barbora , Vlach Jiří , Junková Petra , Karamonová Ludmila , Blažková Martina , Fukal Ladislav
Published: April 2017

In the last decade, strains of the genera
Franconibacter
and
Siccibacter
have been misclassified as first
Enterobacter
and later
Cronobacter
. Because
Cronobacter
is a serious foodborne pathogen that affects premature neonates and elderly individuals, such misidentification may not only falsify epidemiological statistics but also lead to tests of powdered infant formula or other foods giving false results. Currently, the main ways of identifying
Franconibacter
and
Siccibacter
strains are by biochemical testing or by sequencing of the
fusA
gene as part of
Cronobacter
multilocus sequence typing (MLST), but in relation to these strains the former is generally highly difficult and unreliable while the latter remains expensive. To address this, we developed a fast, simple, and most importantly, reliable method for
Franconibacter
and
Siccibacter
identification based on intact-cell matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS). Our method integrates the following steps: data preprocessing using mMass software; principal-component analysis (PCA) for the selection of mass spectrum fingerprints of
Franconibacter
and
Siccibacter
strains; optimization of the Biotyper database settings for the creation of main spectrum projections (MSPs). This methodology enabled us to create an in-house MALDI MS database that extends the current MALDI Biotyper database by including
Franconibacter
and
Siccibacter
strains. Finally, we verified our approach using seven previously unclassified strains, all of which were correctly identified, thereby validating our method.

IMPORTANCE
We show that the majority of methods currently used for the identification of
Franconibacter
and
Siccibacter
bacteria are not able to properly distinguish these strains from those of
Cronobacter
. While sequencing of the
fusA
gene as part of
Cronobacter
MLST remains the most reliable such method, it is highly expensive and time-consuming. Here, we demonstrate a cost-effective and reliable alternative that correctly distinguishes between
Franconibacter
,
Siccibacter
, and
Cronobacter
bacteria and identifies
Franconibacter
and
Siccibacter
at the species level. Using intact-cell MALDI-TOF MS, we extend the current MALDI Biotyper database with 11
Franconibacter
and
Siccibacter
MSPs. In addition, the use of our approach is likely to lead to a more reliable identification scheme for
Franconibacter
and
Siccibacter
strains and, consequently, a more trustworthy epidemiological picture of their involvement in disease.
Rational Engineering of a Cold-Adapted α-Amylase from the Antarctic Ciliate Euplotes focardii for Simultaneous Improvement of Thermostability and Catalytic Activity
Authors: Yang Guang , Yao Hua , Mozzicafreddo Matteo , Ballarini Patrizia , Pucciarelli Sandra , Miceli Cristina
Published: April 2017

The α-amylases are endo-acting enzymes that hydrolyze starch by randomly cleaving the 1,4-α-
d
-glucosidic linkages between the adjacent glucose units in a linear amylose chain. They have significant advantages in a wide range of applications, particularly in the food industry. The eukaryotic α-amylase isolated from the Antarctic ciliated protozoon
Euplotes focardii
(
Ef
Amy) is an alkaline enzyme, different from most of the α-amylases characterized so far. Furthermore,
Ef
Amy has the characteristics of a psychrophilic α-amylase, such as the highest hydrolytic activity at a low temperature and high thermolability, which is the major drawback of cold-active enzymes in industrial applications. In this work, we applied site-directed mutagenesis combined with rational design to generate a cold-active
Ef
Amy with improved thermostability and catalytic efficiency at low temperatures. We engineered two
Ef
Amy mutants. In one mutant, we introduced Pro residues on the A and B domains in surface loops. In the second mutant, we changed Val residues to Thr close to the catalytic site. The aim of these substitutions was to rigidify the molecular structure of the enzyme. Furthermore, we also analyzed mutants containing these combined substitutions. Biochemical enzymatic assays of engineered versions of
Ef
Amy revealed that the combination of mutations at the surface loops increased the thermostability and catalytic efficiency of the enzyme. The possible mechanisms responsible for the changes in the biochemical properties are discussed by analyzing the three-dimensional structural model.

IMPORTANCE
Cold-adapted enzymes have high specific activity at low and moderate temperatures, a property that can be extremely useful in various applications as it implies a reduction in energy consumption during the catalyzed reaction. However, the concurrent high thermolability of cold-adapted enzymes often limits their applications in industrial processes. The α-amylase from the psychrophilic Antarctic ciliate
Euplotes focardii
(named
Ef
Amy) is a cold-adapted enzyme with optimal catalytic activity in an alkaline environment. These unique features distinguish it from most α-amylases characterized so far. In this work, we engineered a novel
Ef
Amy with improved thermostability, substrate binding affinity, and catalytic efficiency to various extents, without impacting its pH preference. These characteristics can be considered important properties for use in the food, detergent, and textile industries and in other industrial applications. The enzyme engineering strategy developed in this study may also provide useful knowledge for future optimization of molecules to be used in particular industrial applications.
Improved α-Amylase Production by Dephosphorylation Mutation of CreD, an Arrestin-Like Protein Required for Glucose-Induced Endocytosis of Maltose Permease and Carbon Catabolite Derepression in Aspergillusoryzae
Authors: Tanaka Mizuki , Hiramoto Tetsuya , Tada Hinako , Shintani Takahiro , Gomi Katsuya
Published: April 2017

Aspergillus
oryzae
produces copious amount of amylolytic enzymes, and MalP, a major maltose permease, is required for the expression of amylase-encoding genes. The expression of these genes is strongly repressed by carbon catabolite repression (CCR) in the presence of glucose. MalP is transported from the plasma membrane to the vacuole by endocytosis, which requires the homolog of E6-AP carboxyl terminus ubiquitin ligase HulA, an ortholog of yeast Rsp5. In yeast, arrestin-like proteins mediate endocytosis as adaptors of Rsp5 and transporters. In the present study, we examined the involvement of CreD, an arrestin-like protein, in glucose-induced MalP endocytosis and CCR of amylase-encoding genes. Deletion of
creD
inhibited the glucose-induced endocytosis of MalP, and CreD showed physical interaction with HulA. Phosphorylation of CreD was detected by Western blotting, and two serine residues were determined as the putative phosphorylation sites. However, the phosphorylation state of the serine residues did not regulate MalP endocytosis and its interaction with HulA. Although α-amylase production was significantly repressed by
creD
deletion, both phosphorylation and dephosphorylation mimics of CreD had a negligible effect on α-amylase activity. Interestingly, dephosphorylation of CreD was required for CCR relief of amylase genes that was triggered by disruption of the deubiquitinating enzyme-encoding gene
creB
. The α-amylase activity of the
creB
mutant was 1.6-fold higher than that of the wild type, and the dephosphorylation mimic of CreD further improved the α-amylase activity by 2.6-fold. These results indicate that a combination of the dephosphorylation mutation of CreD and
creB
disruption increased the production of amylolytic enzymes in
A. oryzae
.

IMPORTANCE
In eukaryotes, glucose induces carbon catabolite repression (CCR) and proteolytic degradation of plasma membrane transporters via endocytosis. Glucose-induced endocytosis of transporters is mediated by their ubiquitination, and arrestin-like proteins act as adaptors of transporters and ubiquitin ligases. In this study, we showed that CreD, an arrestin-like protein, is involved in glucose-induced endocytosis of maltose permease and carbon catabolite derepression of amylase gene expression in
Aspergillus
oryzae
. Dephosphorylation of CreD was required for CCR relief triggered by the disruption of
creB
, which encodes a deubiquitinating enzyme; a combination of the phosphorylation-defective mutation of CreD and
creB
disruption dramatically improved α-amylase production. This study shows the dual function of an arrestin-like protein and provides a novel approach for improving the production of amylolytic enzymes in
A. oryzae
.
Yersinia pestis Resists Predation by Acanthamoeba castellanii and Exhibits Prolonged Intracellular Survival
Authors: Benavides-Montaño Javier A. , Vadyvaloo Viveka
Published: April 2017

Plague is a flea-borne rodent-associated zoonotic disease caused by
Yersinia pestis
. The disease is characterized by epizootics with high rodent mortalities, punctuated by interepizootic periods when the bacterium persists in an unknown reservoir. This study investigates the interaction between
Y. pestis
and the ubiquitous soil free-living amoeba (FLA)
Acanthamoeba castellanii
to assess if the bacterium can survive within soil amoebae and whether intracellular mechanisms are conserved between infection of mammalian macrophages and soil amoebae. The results demonstrate that during coculture with amoebae, representative
Y. pestis
strains of epidemic biovars Medievalis, Orientalis, and Antiqua are phagocytized and able to survive within amoebae for at least 5 days. Key
Y. pestis
determinants of the intracellular interaction of
Y. pestis
and phagocytic macrophages, PhoP and the type three secretion system (T3SS), were then tested for their roles in the
Y. pestis
-amoeba interaction. Consistent with a requirement for the PhoP transcriptional activator in the intracellular survival of
Y. pestis
in macrophages, a PhoP mutant is unable to survive when cocultured with amoebae. Additionally, induction of the T3SS blocks phagocytic uptake of
Y. pestis
by amoebae, similar to that which occurs during macrophage infection. Electron microscopy revealed that in
A. castellanii
,
Y. pestis
resides intact within spacious vacuoles which were characterized using lysosomal trackers as being separated from the lysosomal compartment. This evidence for prolonged survival and subversion of intracellular digestion of
Y. pestis
within FLA suggests that protozoa may serve as a protective soil reservoir for
Y. pestis
.

IMPORTANCE
Yersinia pestis
is a reemerging flea-borne zoonotic disease. Sylvatic plague cycles are characterized by an epizootic period during which the disease spreads rapidly, causing high rodent mortality, and an interepizootic period when the bacterium quiescently persists in an unknown reservoir. An understanding of the ecology of
Y. pestis
in the context of its persistence in the environment and its reactivation to initiate a new epizootic cycle is key to implementing novel surveillance strategies to more effectively predict and prevent new disease outbreaks. Here, we demonstrate prolonged survival and subversion of intracellular digestion of
Y. pestis
within a soil free-living amoeba. This suggests the potential role for protozoa as a protective soil reservoir for
Y. pestis
, which may help explain the recrudescence of plague epizootics.
Lack of Methylated Hopanoids Renders the Cyanobacterium Nostoc punctiforme Sensitive to Osmotic and pH Stress
Authors: Garby Tamsyn J. , Matys Emily D. , Ongley Sarah E. , Salih Anya , Larkum Anthony W. D. , Walter Malcolm R. , Summons Roger E. , Neilan Brett A.
Published: April 2017

To investigate the function of 2-methylhopanoids in modern cyanobacteria, the
hpnP
gene coding for the radical
S
-adenosyl methionine (SAM) methylase protein that acts on the C-2 position of hopanoids was deleted from the filamentous cyanobacterium
Nostoc punctiforme
ATCC 29133S. The resulting Δ
hpnP
mutant lacked all 2-methylhopanoids but was found to produce much higher levels of two bacteriohopanepentol isomers than the wild type. Growth rates of the Δ
hpnP
mutant cultures were not significantly different from those of the wild type under standard growth conditions. Akinete formation was also not impeded by the absence of 2-methylhopanoids. The relative abundances of the different hopanoid structures in akinete-dominated cultures of the wild-type and Δ
hpnP
mutant strains were similar to those of vegetative cell-dominated cultures. However, the Δ
hpnP
mutant was found to have decreased growth rates under both pH and osmotic stress, confirming a role for 2-methylhopanoids in stress tolerance. Evidence of elevated photosystem II yield and NAD(P)H-dependent oxidoreductase activity in the Δ
hpnP
mutant under stress conditions, compared to the wild type, suggested that the absence of 2-methylhopanoids increases cellular metabolic rates under stress conditions.

IMPORTANCE
As the first group of organisms to develop oxygenic photosynthesis,
Cyanobacteria
are central to the evolutionary history of life on Earth and the subsequent oxygenation of the atmosphere. To investigate the origin of cyanobacteria and the emergence of oxygenic photosynthesis, geobiologists use biomarkers, the remnants of lipids produced by different organisms that are found in geologic sediments. 2-Methylhopanes have been considered indicative of cyanobacteria in some environmental settings, with the parent lipids 2-methylhopanoids being present in many contemporary cyanobacteria. We have created a
Nostoc punctiforme
Δ
hpnP
mutant strain that does not produce 2-methylhopanoids to assess the influence of 2-methylhopanoids on stress tolerance. Increased metabolic activity in the mutant under stress indicates compensatory alterations in metabolism in the absence of 2-methylhopanoids.
Effect of cardiorespiratory and strength exercises on disease activity in patients with inflammatory rheumatic diseases: a systematic review and meta-analysis
Authors: Sveaas Silje Halvorsen , Smedslund Geir , Hagen Kåre Birger , Dagfinrud Hanne
Published: April 2017

Objective

To investigate the effects of cardiorespiratory and strength exercises on disease activity for patients with inflammatory rheumatic diseases (IRDs).

Design

A systematic review with meta-analysis registered at PROSPERO (CRD42015020004).

Participants

Patients with IRDs.

Data sources

The databases MEDLINE, AMED, Embase and CINAHL were searched from inception up to April 2016.

Eligibility criteria for selecting studies

Trials were included if they were randomised controlled trials of adults with IRDs, comparing the effect of cardiorespiratory and strength exercises with usual care on disease activity and followed the American College of Sports Medicine's exercise recommendations. The primary outcome was disease activity in terms of inflammation, joint damage and symptoms.

Data synthesis

Data were pooled in a random-effect model for all outcomes, and standardised mean differences (SMDs) were calculated. The quality of evidence was evaluated according to the Grading of Recommendations Assessment, Development and Evaluation approach.

Results

Twenty-six trials with a total of 1286 participants were included. There was high to moderate quality evidence, for a small beneficial effect on disease activity scores (0.19 (95% CI 0.05 to 0.33), p<0.01) and joint damage (SMD 0.27 (95% CI 0.07 to 0.46), p<0.01). Furthermore, moderate quality evidence for a small beneficial effect on erythrocyte sedimentation rate (SMD 0.20 (95% CI 0.0 to 0.39), p=0.04) and for no effect on C reactive protein (SMD −0.14 (95% CI −0.37 to 0.08), p=0.21). Beneficial effects were also seen for symptoms.

Conclusions

The results of this review suggest beneficial effects of exercises on inflammation, joint damage and symptoms in patients with IRDs.
Role of advanced neuroimaging, fluid biomarkers and genetic testing in the assessment of sport-related concussion: a systematic review
Authors: McCrea Michael , Meier Timothy , Huber Daniel , Ptito Alain , Bigler Erin , Debert Chantel T , Manley Geoff , Menon David , Chen Jen-Kai , Wall Rachel , Schneider Kathryn J , McAllister Thomas
Published: April 2017

Objective

To conduct a systematic review of published literature on advanced neuroimaging, fluid biomarkers and genetic testing in the assessment of sport-related concussion (SRC).

Data sources

Computerised searches of Medline, PubMed, Cumulative Index to Nursing and Allied Health Literature (CINAHL), PsycINFO, Scopus and Cochrane Library from 1 January 2000 to 31 December 2016 were done. There were 3222 articles identified.

Study selection

In addition to medical subject heading terms, a study was included if (1) published in English, (2) represented original research, (3) involved human research, (4) pertained to SRC and (5) involved data from neuroimaging, fluid biomarkers or genetic testing collected within 6 months of injury. Ninety-eight studies qualified for review (76 neuroimaging, 16 biomarkers and 6 genetic testing).

Data extraction

Separate reviews were conducted for neuroimaging, biomarkers and genetic testing. A standardised data extraction tool was used to document study design, population, tests employed and key findings. Reviewers used a modified quality assessment of studies of diagnostic accuracy studies (QUADAS-2) tool to rate the risk of bias, and a modified Grading of Recommendations Assessment, Development, and Evaluation (GRADE) system to rate the overall level of evidence for each search.

Data synthesis

Results from the three respective reviews are compiled in separate tables and an interpretive summary of the findings is provided.

Conclusions

Advanced neuroimaging, fluid biomarkers and genetic testing are important research tools, but require further validation to determine their ultimate clinical utility in the evaluation of SRC. Future research efforts should address current gaps that limit clinical translation. Ultimately, research on neurobiological and genetic aspects of SRC is predicted to have major translational significance to evidence-based approaches to clinical management of SRC, much like applied clinical research has had over the past 20 years.
What is the difference between comprehensive and selective primary health care? Evidence from a five-year longitudinal realist case study in South Australia
Authors: Baum Fran , Freeman Toby , Lawless Angela , Labonte Ronald , Sanders David
Published: April 2017

Background

Since the WHO’s Alma Ata Declaration on Primary Health Care (PHC) there has been debate about the advisability of adopting comprehensive or selective PHC. Proponents of the latter argue that a more selective approach will enable interim gains while proponents of a comprehensive approach argue that it is needed to address the underlying causes of ill health and improve health outcomes sustainably.

Methods

This research is based on four case studies of government-funded and run PHC services in Adelaide, South Australia. Program logic models were constructed from interviews and workshops. The initial model represented relatively comprehensive service provision in 2010. Subsequent interviews in 2013 permitted the construction of a selective PHC program logic model following a series of restructuring service changes.

Results

Comparison of the PHC service program logic models before and after restructuring illustrates the changes to the operating context, underlying mechanisms, service qualities, activities, activity outcomes and anticipated community health outcomes. The PHC services moved from focusing on a range of community, group and individual clinical activities to a focus on the management of people with chronic disease. Under the more comprehensive model, activities were along a continuum of promotive, preventive, rehabilitative and curative. Under the selective model, the focus moved to rehabilitative and curative with very little other activities.

Conclusion

The study demonstrates the difference between selective and comprehensive approaches to PHC in a rich country setting and is useful in informing debates on PHC especially in the context of the Sustainable Development Goals.
Essential components of integrated care for youth with mental health and addiction needs: protocol for a scoping review
Authors: Settipani Cara A , Cleverley Kristin , Hawke Lisa D , Rice Maureen , Henderson Joanna L
Published: April 2017

Introduction

Efforts to move towards integrated care have been met with increased interest and enthusiasm in recent years given the potential to improve care and population health while containing costs. However, there is a need to better understand community-based integrated care approaches for youth with mental health and/or addiction concerns to guide future implementation efforts and develop a set of standards for key components. The objectives of this scoping review are to: (1) identify the populations, settings, service providers, interventions, infrastructure and care coordination methods that have been included in integrated care for youth with mental health and/or addiction needs and (2) identify constructs that have been measured and evaluated (eg, outcomes, engagement) in the context of youth integrated care.

Methods and analysis

Seven electronic databases and several grey literature sources will be searched for material from 2001 to 2016. Inclusion criteria will be broad with respect to type of work, as we will include all types of research studies as well as non-research studies that provide information relevant to characteristics and constructs measured in the context of integrated care for youth mental health. Titles and abstracts will be independently screened for eligibility by two raters using inclusion criteria. Full-text articles will then be accessed and independently screened for inclusion. A formal data extraction method will be employed, enabling synthesis of results in quantitative and qualitative formats.

Ethics and dissemination

Results will be widely disseminated to various stakeholders to inform implementation and research efforts. Findings will also launch a Delphi method study leading to the development of an assessment tool for youth mental health services integration. This review does not require ethics approval.
The Value of a Surgical Assessment Unit Ultrasound Facility
Authors: Lai Wesley , Gutteridge Catherine , Regan Alicia , Lambert Anthony
Published: April 2017

Ultrasound scan (USS) is a common and important mode of investigation for emergency surgical admissions. Delay in investigation often leads to delayed diagnosis and treatment, and possible extended length of stay (LOS), which has clinical, cost and service provision implications. We aim to investigate the clinical impact on patient care and the cost-effectiveness of a pilot Surgical Assessment Unit (SAU) USS facility. We performed a retrospective data collection on 100 consecutive SAU inpatients who had an USS investigation on the ward since the introduction of the facility, matched by 100 consecutive SAU inpatients who had an USS in the radiology department before the pilot study. Results of the audit show SAU USS has a reduced mean LOS by 1.44 days compared to departmental USS, and led to more same day discharge than departmental USS (20 vs. 5), thus avoiding unnecessary overnight stay. It also significantly reduced mean waiting time from admission to investigation by 5.21 hours, which can be translated into improved patient and staff satisfaction. All these findings are both statistically and clinically significant. The estimated cost of each SAU USS is comparable to the average departmental USS (£29.71 vs. £30.80). Using the average cost of an excess bed day = £273, SAU USS has produced an estimated saving of £394.72/patient. This does not include saved opportunistic costs such as prevented elective operation cancellations, fines incurred from surgery waiting time/A+E breaches etc. To conclude SAU USS has a significant positive impact on patient care in surgical admissions by reducing LOS and investigation waiting time, as well as facilitating same day discharge.
SWITCH: Al Wakra Hospital Journey to 90% Hand Hygiene Practice Compliance, 2011 – 2015
Authors: Visan Feah Altura - , Zakaria Almunzer , Castro Jenalyn , Alhasanat Omar , Ismail Khalil Al , Ansari Naser Al , Hamed Manal
Published: April 2017

Hand Hygiene is the cheapest and simplest way to prevent the spread of infection, however international compliance is below than 40% (WHO, 2009). In the experience of Al Wakra Hospital, the improvement in hand hygiene compliance highlighted not just interventions towards training and education but also behavioral motivation and physical allocations of hand hygiene appliances and equipment.

Through motivating the behavioral, emotional, physical and intellectual dimensions of the different healthcare worker professions, hand hygiene compliance has increased from 60.78% in 2011 to 94.14% by the end of December 2015. It took 25 months of continuous and collaborative work with different healthcare workers to reach the 90% hand hygiene target.

“Together, we have reached our goals and together we fight against infections! Because we always strive for excellence in everything we do – that is our vision here in Al Wakra Hospital.”
Reducing Central Venous Catheter Use in Peripheral Blood Stem Cell Donation: Quality Improvement Report
Authors: Ghazi Samer , Alaskar Ahmed , Alzahrani Mohsen , Damlaj Moussab , Abuelgasim Khadega A , Gmati Giamal , Alshami Mona , Alshammary Salman , Al-Surimi Khaled , Salama Hind , Alhejazi Ayman , Jazieh Abdul-Rahman
Published: April 2017

Peripheral blood stem cell (PBSC) collection from donors through apheresis has become the main source of stem cells for hematopoietic stem cell transplantation. This procedure requires a high blood flow venous access. A peripheral venous catheter (PVC), compared to a central venous catheter (CVC), is considered to provide safer venous access. However, initially at our institution, King Abdul-Aziz Medical City - Riyadh, a CVC was frequently used (72%). A quality improvement multidisciplinary team has been formed to conduct a systematic quality performance analysis to evaluate the current process of collecting donor PBSCs with the aim to reduce CVC use to less than the international benchmark (20%). A quality improvement methodology, rapid cycles of plan-do-study-act (PDSA), was used to test a set of initiatives. An Intravenous (IV) team assessed the donor's venous access and inserted an appropriate PVC when feasible. This project ran over 16 months with 42 adult donors undergoing PBSC collection. During the first PDSA cycle, 1 CVC was inserted for every 4 donors. In the second PDSA cycle, 1 CVC was inserted for every 8 apheresis donations. In the third PDSA cycle, no CVC was used for 30 apheresis donations. The targeted stem cell dose was collected successfully in one apheresis session in all donors assigned for PVC access with no complications. A significant reduction of CVC use from 72% to 0% was achieved. This quality improvement project demonstrated that a successful apheresis procedure can be achieved easily and safely in the majority of PBSC donors preventing the potential adverse events associated with CVCs. The interdisciplinary collaboration between the IV team, apheresis and clinical hematology teams was paramount to optimize the safe care of donors.