bluuebunny/crossref_metadata_2025 huggingface dataset:
An international survey on the role of the hospital transfusion committee
Authors: Yazer Mark H. , Lozano Miguel , Fung Mark , Kutner Jose , Murphy Michael F. , Oveland Apelseth Torunn , Pogłód Ryszard , Selleng Kathleen , Tinmouth Alan , Wendel Silvano , Yahalom Vered ,   on behalf of the Biomedical Excellence for Safer Transfusion (BEST) Collaborative
Published: February 2017

BACKGROUND

Hospital transfusion committees (HTCs) can oversee all aspects of transfusion practice at a hospital. This survey sought to identify which quality variables were being reported at HTCs around the world.

STUDY DESIGN AND METHODS

A working party composed of members of the Biomedical Excellence for Safer Transfusion (BEST) collaborative developed a survey of quality variables that could be potentially presented at HTC meetings. The survey was electronically sent to all BEST members who were encouraged to complete it if they were active on an HTC and to send it to other colleagues with similar experience. An expert panel was convened to determine which quality variables are the most important for review at HTC meetings.

RESULTS

There were 121 respondents; the majority were from Europe (52%), Asia (19%), or North America (19%). Most respondents (68%) were at university hospitals. Of the 117 (97%) respondents with an HTC, the committee most often met quarterly (42%) and reviewed transfusion reactions (79%) and risk management–reported events (52%). The HTCs most commonly included transfusion medicine physicians, anesthesiologists, and other physicians who regularly transfuse blood products. Some of the most commonly reported quality variables included number of blood products transfused, wasted, and expired and the number of improperly labeled specimens. The expert panel analysis revealed that some variables that were deemed important were not being frequently reported at HTCs.

CONCLUSION

There is variability in the variables being reported at HTCs around the world with some important variables not frequently reported.
p38 mitogen‐activated protein kinase regulates mitochondrial function and microvesicle release in riboflavin‐ and ultraviolet light–treated apheresis platelet concentrates
Authors: Chen Zhongming , Schubert Peter , Bakkour Sonia , Culibrk Brankica , Busch Michael P. , Devine Dana V.
Published: February 2017

BACKGROUND

Biochemical analyses of mechanisms triggered in platelets (PLTs) upon pathogen inactivation (PI) are crucial to further understand the impact of PI on PLT functionality and, subsequently, quality.

STUDY DESIGN AND METHODS

PLT concentrates (PCs) were split into four small illumination bags: 1) untreated control, 2) treated with riboflavin and ultraviolet light (RF/UV), and spiked with 3) solvent control dimethyl sulfoxide and 4) p38 mitogen‐activated protein kinase (MAPK) inhibitor SB203580 before RF/UV treatment. Flow cytometry was used to monitor PLT mitochondrial potential (ΔΨm); generation of intracellular reactive oxygen species (ROS); and release of microvesicles (MVs), mitochondria (MT), and MVs containing MT (MVs/MT). Quantitative polymerase chain reaction (qPCR) was used to quantify extracellular mitochondrial DNA (mtDNA). Translocation of selected mitochondrial proteins was analyzed in subcellular fractions by immunoblot.

RESULTS

RF/UV treatment triggered an increased mitochondrial translocation of both Bax and Bid (p < 0.05, Day 7) and cytochromecrelease (p < 0.01, Day 7), loss of ΔΨm(p < 0.05, Day 5 and Day 7), and ROS generation (p < 0.01, Day 5 and Day 7) in PCs compared to the untreated control during storage. These PI‐triggered changes were inhibited by SB203580 (p < 0.05). The release of MVs, MT, and MVs/MT was increased upon the RF/UV treatment during storage (p < 0.05) and, with the exception of MT, the release was decreased by the inhibitor (p < 0.05). qPCR analysis showed that RF/UV does not trigger mtDNA release during storage.

CONCLUSION

These findings further our understanding of mechanisms in PLTs initiated by the RF/UV treatment, demonstrating that this treatment induces p38 MAPK‐dependent mitochondrial signaling and MV release in apheresis PCs.
Prioritizing of bacterial infections transmitted through substances of human origin in Europe
Authors: Domanović Dragoslav , Cassini Alessandro , Bekeredjian‐Ding Isabelle , Bokhorst Arlinke , Bouwknegt Martijn , Facco Giuseppina , Galea George , Grossi Paolo , Jashari Ramadan , Jungbauer Christoph , Marcelis Jan , Raluca‐Siska Ioana , Andersson‐Vonrosen Inger , Suk Jonathan E.
Published: February 2017

BACKGROUND

Bacteria are the pathogens most frequently transmitted through substances of human origin (SoHO). The European Centre for Disease Prevention and Control (ECDC) organized an expert consultation, with the objective of developing a priority list of bacterial pathogens transmissible via SoHO. The list will be used to further assess risks and determine appropriate preventive measures.

STUDY DESIGN AND METHODS

The 14 most frequently SoHO‐transmitted bacteria identified through a scoping literature review were then prioritized during an expert workshop through a methodology based on multicriteria decision analysis. The selection of the prioritization method was based upon an ECDC framework for best practices in conducting risk‐ranking exercises. Three transmission pathways, blood and blood components, tissues and cells, and organs, were considered in the ranking exercise.

RESULTS

According to the ranking score (RS), bacteria were organized within each SoHO pathway into one of four risk tiers: Tier 1 (RS ≥ 0.70), Tier 2 (RS = 0.60‐0.69), Tier 3 (RS = 0.40‐0.59), or Tier 4 (RS < 0.40). The most consistently identified pathogens in the highest risk Tiers 1 and 2 of all three pathways were: Staphylococcus aureus, Klebsiella spp., Escherichia coli, β‐hemolytic streptococci, Pseudomonas spp., and Acinetobacter spp.

CONCLUSION

Six bacteria were defined as being of the highest priority in respect of the threat to the safety of SoHO and will be the subject of subsequent in‐depth risk assessments to be conducted by ECDC to identify measures to mitigate the risk posed by these bacteria.
Clinical effectiveness of platelets in additive solution treated with two commercial pathogen‐reduction technologies
Authors: Rebulla Paolo , Vaglio Stefania , Beccaria Francesco , Bonfichi Maurizio , Carella Angelo , Chiurazzi Federico , Coluzzi Serelina , Cortelezzi Agostino , Gandini Giorgio , Girelli Gabriella , Graf Maria , Isernia Paola , Marano Giuseppe , Marconi Maurizio , Montemezzi Rachele , Olivero Barbara , Rinaldi Marianna , Salvaneschi Laura , Scarpato Nicola , Strada Paolo , Milani Silvano , Grazzini Giuliano
Published: February 2017

BACKGROUND

Two noninferiority, randomized, controlled trials were conducted in parallel comparing the safety and efficacy of platelets treated with Intercept or Mirasol pathogen‐reduction technologies versus standard platelets.

STUDY DESIGN AND METHODS

The primary endpoint was the percentage of hematology patients who developed World Health Organization Grade 2 or greater bleeding. A noninferiority margin of 11% was chosen based on expected Grade 2 or greater bleeding in 20% of controls. The study was closed for financial restrictions before reaching the planned sample size of 828 patients, and an intention‐to‐treat analysis was conducted on 424 evaluable patients.

RESULTS

In the Intercept trial (113 treated vs. 115 control patients), the absolute risk difference in Grade 2 or greater bleeding was 6.1%, with an upper one‐sided 97.5% confidence limit of 19.2%. The absolute risk difference in the Mirasol trial (99 treated vs. 97 control patients) was 4.1%, and the upper one‐sided 97.5% confidence limit was 18.4%. Neither absolute risk difference was statistically significant. In both trials, posttransfusion platelet count increments were significantly lower in treated versus control patients. Mean blood component use in treated patients versus controls was 54% higher (95% confidence interval, 36%‐74%; Intercept) and 34% higher (95% confidence interval, 16%‐54%; Mirasol) for platelets and 23% higher (95% confidence interval, 8%‐39%; Intercept) and 32% higher (95% confidence interval, 10%‐57%; Mirasol) for red blood cells. Unexpected reactions and adverse events were not reported. Mortality did not differ significantly between treated and control patients.

CONCLUSION

Although conclusions on noninferiority could not be drawn due to low statistical power, the study provides additional information on the safety and efficacy of pathogen‐reduced platelets treated with two commercial pathogen‐reduction technologies.
Pathogen inactivation treatment of plasma and platelet concentrates and their predicted functionality in massive transfusion protocols
Authors: Arbaeen Ahmad F. , Schubert Peter , Serrano Katherine , Carter Cedric J. , Culibrk Brankica , Devine Dana V.
Published: February 2017

BACKGROUND

Trauma transfusion packages for hemorrhage control consist of red blood cells, plasma, and platelets at a set ratio. Although pathogen reduction improves the transfusion safety of platelet and plasma units, there is an associated reduction in quality. This study aimed to investigate the impact of riboflavin/ultraviolet light‐treated plasma or platelets in transfusion trauma packages composed of red blood cell, plasma, and platelet units in a ratio of 1:1:1 in vitro by modeling transfusion scenarios for trauma patients and assessing function by rotational thromboelastometry.

STUDY DESIGN AND METHODS

Pathogen‐reduced or untreated plasma and buffy coat platelet concentrate units produced in plasma were used in different combinations with red blood cells in trauma transfusion packages. After reconstitution of these packages with hemodiluted blood, the hemostatic functionality was analyzed by rotational thromboelastometry.

RESULTS

Hemostatic profiles of pathogen‐inactivated buffy coat platelet concentrate and plasma indicated decreased activity compared with their respective controls. Reconstitution of hemodiluted blood (hematocrit = 20%) with packages that contained treated or nontreated components resulted in increased alpha and maximum clot firmness and enhanced clot‐formation time. Simulating transfusion scenarios based on 30% blood replacement with a transfusion trauma package resulted in a nonsignificant difference in rotational thromboelastometry parameters between packages containing treated and nontreated blood components (p ≥ 0.05). Effects of pathogen inactivation treatment were evident when the trauma package percentage was 50% or greater and contained both pathogen inactivation‐treated plasma and buffy coat platelet concentrate.

CONCLUSION

Rotational thromboelastometry investigations suggest that there is relatively little impact of pathogen inactivation treatment on whole blood clot formation unless large amounts of treated components are used.
Characterization of hepatitis B virus (HBV) preS/S gene mutations in blood donors with occult HBV infection in the Baoji area of North China
Authors: Liao Hao , Liu Yan , Chen Jianhong , Ding Weiping , Li Xiaodong , Xu Zhihui , Yang Yuan , Chen Rongjuan , Si Lanlan , Xu Xiaoyuan , Guo Jianxun , Xu Dongping
Published: February 2017

BACKGROUND

Occult hepatitis B virus (HBV) infection (OBI) in blood donors was investigated in the Baoji area of North China, and OBI‐related viral mutations in donors were characterized.

STUDY DESIGN AND METHODS

In total, 110,843 blood donor samples that were consecutively collected from December 2011 to March 2015 at the Baoji Blood Center were examined. Hepatitis B surface antigen‐negative and HBV DNA‐positive OBI samples were amplified for sequence analysis of OBI‐related mutations in the HBV preS/S region. HBV genomes from 108 adult patients with chronic hepatitis B from North China were used as controls.

RESULTS

OBI was detected in 60 (1:1847) individual blood donors. All OBI samples were negative for hepatitis B e‐antigen, and 55 were positive for anti‐hepatitis B core antigen. The preS/S genes were successfully sequenced for 43 OBI samples. OBI‐related S gene mutations in the major hydrophilic region were detected more frequently in blood donors with OBI than that in controls (51.16 vs. 12.96%; p < 0.01). Specifically, the incidence of five OBI‐related major hydrophilic region mutations (sS117T, sT118K, sT131N, sT134Y/L, and sD144E) was significantly higher in blood donors with OBI than in controls. In addition, the coexistence of multiple OBI‐related mutations in the major hydrophilic region was detected more frequently in donors than in controls (30.23 vs. 1.85%; p < 0.01), and preS deletions greater than 33 base pairs also were detected more frequently in blood donors with OBI than in controls.

CONCLUSION

OBI in blood donors should be addressed attentively in the Baoji area of North China, and HBV preS/S gene mutations may play an important role in OBI prevalence in the area.
Human platelet antigen (HPA)‐specific immunoglobulin M antibodies in neonatal alloimmune thrombocytopenia can inhibit the binding of HPA‐specific immunoglobulin G antibodies
Authors: Hopkins Matthew , Lucas Geoff , Calvert Anthony , Bendukidze Nina , Green Frances , Kotecha Krishna , Poles Anthony
Published: February 2017

BACKGROUND

A term baby with unexplained thrombocytopenia and a platelet (PLT) count of 14 × 109/L (maternal PLT count was 200 × 109/L) was investigated for neonatal alloimmune thrombocytopenia.

STUDY DESIGN AND METHODS

Serologic investigations were performed using the PLT immunofluorescence test (PIFT), monoclonal antibody immobilization of PLT antigens (MAIPA), and a bead‐based assay (BBA) with maternal sera taken up to 56 days postdelivery. One serum sample was also separated into “immunoglobulin (Ig)M‐rich” and “IgM‐depleted” fractions and tested for PLT‐specific antibodies. The family was genotyped for HPA.

RESULTS

HPA‐3a–specific IgM antibodies were detected in the PIFT and confirmed in the BBA. PLT‐specific IgG HPA‐3a antibodies were not detected in the MAIPA assay and BBA in the initial sample but were detected in both techniques in subsequent serum samples. Testing of IgM‐rich and IgM‐depleted fractions in the MAIPA assay revealed that IgG antibody binding of the IgM‐depleted fraction was inhibited by approximately 50% when it was reconstituted with the IgM‐rich fraction suggesting that the IgM antibodies blocked the binding of the IgG antibodies. This effect was not observed when the IgM‐depleted fraction or untreated serum was diluted with elution buffer. Incompatibility for HPA‐3 was identified between the mother and the infant. The infant received one HPA‐1a, −5b negative neonatal PLT transfusion, and one random PLT transfusion, with satisfactory outcomes. Both units were later found to be HPA‐3b3b.

CONCLUSION

HPA‐3a IgM antibodies can inhibit PLT‐specific HPA‐3a IgG antibodies in the MAIPA assay.
Rural habitat as risk factor for hepatitis E virus seroconversion in HIV‐infected patients: A prospective longitudinal study
Authors: Rivero‐Juarez A. , Cuenca‐Lopez F. , Martinez‐Peinado A. , Camacho A. , Real L. M. , Frias M. , Gordon A. , Cantisán S. , Torre‐Cisneros J. , Pineda J. A. , Rivero A.
Published: February 2017

Our objective was to determine the incidence and clinical manifestations of acute hepatitis E virus (HEV) in HIV‐infected patients. A prospective longitudinal study including HIV‐infected HEV‐seronegative patients was conducted; HEV seroconversion (to IgG and/or IgM) was the main outcome variable. All patients were tested for HEV antibodies every 3–6 months. For patients who developed HEV seroconversion, a data collection protocol was followed to identify associated clinical manifestations and analytical alterations. A total of 627 patients (89.9%) were followed during a median of 11.96 months (IQR: 8.52–14.52 months) and formed the study population. Forty‐one patients developed detectable anti‐HEV antibodies (7.2 cases per 100 patients/year). Our study found a high incidence of HEV in HIV‐infected patients in southern Spain strongly associated with a rural habitat.
Effect of Thermal Properties of a Coated Elastohydrodynamic Lubrication Line Contact Under Various Slide-to-Roll Ratios
Authors: Liu Huaiju , Zhu Caichao , Gu Zonglin , Wang Zhanjiang , Tang Jinyuan
Published: March 2017

A numerical thermal elastohydrodynamic lubrication (EHL) model is developed for coated line contacts by considering both the mechanical properties and the thermal properties of the coating and the substrate. The temperature fields within the oil film and within the solids are solved by deriving the energy equations for the solids and the oil film. Heat continuity conditions are satisfied at the interfaces between the solids and the oil film, and the coating/substrate interfaces. Effects of the slide-to-roll ratio (SR), the thermal conductivities of the coating bodies, and the oil film on temperature fields are studied.
Determination of an optimal organ set to implement deformations to support four‐dimensional dose calculations in radiation therapy planning
Authors: Soofi Wafa , Starkschall George , Britton Keith , Vedam Sastry
Published: March 2017

Surface‐based deformable image registration to generate a four‐dimensional (4D) dose calculation in radiation treatment planning requires the selection of a set of organ contours representing a basis set from which to generate anatomic deformation. The purpose of the present work was to determine the optimal set of organs needed to generate a basis set for deformation in treatment planning for thoracic tumors, such that the required computations are minimized, but that dose accuracy is high. Using retrospectively reviewed records and a deformable model algorithm in a research version of a commercial radiation treatment planning system, we calculated 4D dose distributions based on treatment plans for 10 patients with thoracic tumors. Various combinations of organs (total lungs, heart, spinal cord, external body surface) were used to generate the basis set used in the calculations for deformations. The external surface contour did not have a noticeable effect on the dose calculation. Total lung, heart, and spinal cord together provided an adequate set of deformation organs to generate accurate dose deformations. The magnitude of the calculated dose differences had no obvious relationship to tumor parameters, including site, histologic type, disease stage, extent of motion, or degree of centralization.

PACS numbers: 87.55.D‐, 87.55.dk, 87.55.kh
Differential contribution of APP metabolites to early cognitive deficits in a TgCRND8 mouse model of Alzheimer’s disease
Authors: Hamm Valentine , Héraud Céline , Bott Jean-Bastien , Herbeaux Karine , Strittmatter Carole , Mathis Chantal , Goutagny Romain
Published: February 2017

Aβ is unable to account for all aspects of cognitive impairments in a pre-plaque mouse model of Alzheimer’s disease.
Efficient oxygen reduction catalysis by subnanometer Pt alloy nanowires
Authors: Jiang Kezhu , Zhao Dandan , Guo Shaojun , Zhang Xu , Zhu Xing , Guo Jun , Lu Gang , Huang Xiaoqing
Published: February 2017

Highly efficient oxygen reduction electrocatalysis was enabled by a new class of subnanometer Pt alloy nanowires.
Mitochondrial metabolic regulation by GRP78
Authors: Prasad Manoj , Pawlak Kevin J. , Burak William E. , Perry Elizabeth E. , Marshall Brendan , Whittal Randy M. , Bose Himangshu S.
Published: February 2017

ER chaperone GRP78 is the central regulator of adrenals and gonadal steroid synthesis for mammalian survival.
Electrostatic cloaking of surface structure for dynamic wetting
Authors: Nita Satoshi , Do-Quang Minh , Wang Jiayu , Chen Yu-Chung , Suzuki Yuji , Amberg Gustav , Shiomi Junichiro
Published: February 2017

Hindrance of dynamic wetting due to surface microstructures can be deactivated by applying electric fields.
The checkpoint for agonist selection precedes conventional selection in human thymus
Authors: Verstichel Greet , Vermijlen David , Martens Liesbet , Goetgeluk Glenn , Brouwer Margreet , Thiault Nicolas , Van Caeneghem Yasmine , De Munter Stijn , Weening Karin , Bonte Sarah , Leclercq Georges , Taghon Tom , Kerre Tessa , Saeys Yvan , Van Dorpe Jo , Cheroutre Hilde , Vandekerckhove Bart
Published: February 2017

Immature thymocytes are diverted to the agonist selection pathway before selection of conventional T cells.
Tracking the fate and origin of clinically relevant adoptively transferred CD8
+
T cells in vivo
Authors: Chapuis Aude G. , Desmarais Cindy , Emerson Ryan , Schmitt Thomas M. , Shibuya Kendall C. , Lai Ivy P. , Wagener Felecia , Chou Jeffrey , Roberts Ilana M. , Coffey David G. , Warren Edus H. , Robins Harlan , Greenberg Philip D. , Yee Cassian
Published: February 2017

Long-term tumor control originates from very rare T cells with proliferative and/or survival advantages.
Engineering Cyanobacterial Cell Morphology for Enhanced Recovery and Processing of Biomass
Authors: Jordan Adam , Chandler Jenna , MacCready Joshua S. , Huang Jingcheng , Osteryoung Katherine W. , Ducat Daniel C.
Published: February 2017

Cyanobacteria are emerging as alternative crop species for the production of fuels, chemicals, and biomass. Yet, the success of these microbes depends on the development of cost-effective technologies that permit scaled cultivation and cell harvesting. Here, we investigate the feasibility of engineering cell morphology to improve biomass recovery and decrease energetic costs associated with lysing cyanobacterial cells. Specifically, we modify the levels of Min system proteins in
Synechococcus elongatus
PCC 7942. The Min system has established functions in controlling cell division by regulating the assembly of FtsZ, a tubulin-like protein required for defining the bacterial division plane. We show that altering the expression of two FtsZ-regulatory proteins, MinC and Cdv3, enables control over cell morphology by disrupting FtsZ localization and cell division without preventing continued cell growth. By varying the expression of these proteins, we can tune the lengths of cyanobacterial cells across a broad dynamic range, anywhere from an ∼20% increased length (relative to the wild type) to near-millimeter lengths. Highly elongated cells exhibit increased rates of sedimentation under low centrifugal forces or by gravity-assisted settling. Furthermore, hyperelongated cells are also more susceptible to lysis through the application of mild physical stress. Collectively, these results demonstrate a novel approach toward decreasing harvesting and processing costs associated with mass cyanobacterial cultivation by altering morphology at the cellular level.

IMPORTANCE
We show that the cell length of a model cyanobacterial species can be programmed by rationally manipulating the expression of protein factors that suppress cell division. In some instances, we can increase the size of these cells to near-millimeter lengths with this approach. The resulting elongated cells have favorable properties with regard to cell harvesting and lysis. Furthermore, cells treated in this manner continue to grow rapidly at time scales similar to those of uninduced controls. To our knowledge, this is the first reported example of engineering the cell morphology of cyanobacteria or algae to make them more compatible with downstream processing steps that present economic barriers to their use as alternative crop species. Therefore, our results are a promising proof-of-principle for the use of morphology engineering to increase the cost-effectiveness of the mass cultivation of cyanobacteria for various sustainability initiatives.
Metatranscriptomic Profiling Reveals Linkages between the Active Rumen Microbiome and Feed Efficiency in Beef Cattle
Authors: Li Fuyong , Guan Le Luo
Published: February 2017

Exploring compositional and functional characteristics of the rumen microbiome can improve the understanding of its role in rumen function and cattle feed efficiency. In this study, we applied metatranscriptomics to characterize the active rumen microbiomes of beef cattle with different feed efficiencies (efficient,
n
= 10; inefficient,
n
= 10) using total RNA sequencing. Active bacterial and archaeal compositions were estimated based on 16S rRNAs, and active microbial metabolic functions including carbohydrate-active enzymes (CAZymes) were assessed based on mRNAs from the same metatranscriptomic data sets. In total, six bacterial phyla (
Proteobacteria
,
Firmicutes
,
Bacteroidetes
,
Spirochaetes
,
Cyanobacteria
, and
Synergistetes
), eight bacterial families (
Succinivibrionaceae
,
Prevotellaceae
,
Ruminococcaceae
,
Lachnospiraceae
,
Veillonellaceae
,
Spirochaetaceae
,
Dethiosulfovibrionaceae
, and
Mogibacteriaceae
), four archaeal clades (
Methanomassiliicoccales
,
Methanobrevibacter ruminantium
,
Methanobrevibacter gottschalkii
, and
Methanosphaera
), 112 metabolic pathways, and 126 CAZymes were identified as core components of the active rumen microbiome. As determined by comparative analysis, three bacterial families (
Lachnospiraceae
,
Lactobacillaceae
, and
Veillonellaceae
) tended to be more abundant in low-feed-efficiency (inefficient) animals (
P
< 0.10), and one archaeal taxon (
Methanomassiliicoccales
) tended to be more abundant in high-feed-efficiency (efficient) cattle (
P
< 0.10). Meanwhile, 32 microbial metabolic pathways and 12 CAZymes were differentially abundant (linear discriminant analysis score of >2 with a
P
value of <0.05) between two groups. Among them, 30 metabolic pathways and 11 CAZymes were more abundant in the rumen of inefficient cattle, while 2 metabolic pathways and 1 CAZyme were more abundant in efficient animals. These findings suggest that the rumen microbiomes of inefficient cattle have more diverse activities than those of efficient cattle, which may be related to the host feed efficiency variation.

IMPORTANCE
This study applied total RNA-based metatranscriptomics and showed the linkage between the active rumen microbiome and feed efficiency (residual feed intake) in beef cattle. The data generated from the current study provide fundamental information on active rumen microbiome at both compositional and functional levels, which serve as a foundation to study rumen function and its role in cattle feed efficiency. The findings that the active rumen microbiome may contribute to variations in feed efficiency of beef cattle highlight the possibility of enhancing nutrient utilization and improve cattle feed efficiency through modification of rumen microbial functions.
A Cyanide-Induced 3-Cyanoalanine Nitrilase in the Cyanide-Assimilating Bacterium Pseudomonas pseudoalcaligenes Strain CECT 5344
Authors: Acera Felipe , Carmona María Isabel , Castillo Francisco , Quesada Alberto , Blasco Rafael
Published: February 2017

Pseudomonas pseudoalcaligenes
CECT 5344 is a bacterium able to assimilate cyanide as a sole nitrogen source. Under this growth condition, a 3-cyanoalanine nitrilase enzymatic activity was induced. This activity was encoded by
nit4
, one of the four nitrilase genes detected in the genome of this bacterium, and its expression in
Escherichia coli
enabled the recombinant strain to fully assimilate 3-cyanoalanine.
P. pseudoalcaligenes
CECT 5344 showed a weak growth level with 3-cyanoalanine as the N source, unless KCN was also added. Moreover, a
nit4
knockout mutant of
P. pseudoalcaligenes
CECT 5344 became severely impaired in its ability to grow with 3-cyanoalanine and cyanide as nitrogen sources. The native enzyme expressed in
E. coli
was purified up to electrophoretic homogeneity and biochemically characterized. Nit4 seems to be specific for 3-cyanoalanine, and the amount of ammonium derived from the enzymatic activity doubled in the presence of exogenously added asparaginase activity, which demonstrated that the Nit4 enzyme had both 3-cyanoalanine nitrilase and hydratase activities. The
nit4
gene is located downstream of the cyanide resistance transcriptional unit containing
cio1
genes, whose expression levels are under the positive control of cyanide. Real-time PCR experiments revealed that
nit4
expression was also positively regulated by cyanide in both minimal and LB media. These results suggest that this gene cluster including
cio1
and
nit4
could be involved both in cyanide resistance and in its assimilation by
P. pseudoalcaligenes
CECT 5344.

IMPORTANCE
Cyanide is a highly toxic molecule present in some industrial wastes due to its application in several manufacturing processes, such as gold mining and the electroplating industry. The biodegradation of cyanide from contaminated wastes could be an attractive alternative to physicochemical treatment.
P. pseudoalcaligenes
CECT 5344 is a bacterial strain able to assimilate cyanide under alkaline conditions, thus avoiding its volatilization as HCN. This paper describes and characterizes an enzyme (Nit4) induced by cyanide that is probably involved in cyanide assimilation. The biochemical characterization of Nit4 provides a segment for building a cyanide assimilation pathway in
P. pseudoalcaligenes
. This information could be useful for understanding, and hopefully improving, the mechanisms involved in bacterial cyanide biodegradation and its application in the treatment of cyanide-containing wastes.
Experimental Manipulation Shows a Greater Influence of Population than Dietary Perturbation on the Microbiome of Tyrophagus putrescentiae
Authors: Erban Tomas , Ledvinka Ondrej , Nesvorna Marta , Hubert Jan
Published: February 2017

Tyrophagus putrescentiae
is inhabited by bacteria that differ among mite populations (strains) and diets. Here, we investigated how the microbiome and fitness of
T
.
putrescentiae
are altered by dietary perturbations and mite populations. Four
T. putrescentiae
populations, referred to as dog, Koppert, laboratory, and Phillips, underwent a perturbation, i.e., a dietary switch from a rearing diet to two experimental diets. The microbiome was investigated by sequencing the V1-V3 portion of the 16S rRNA gene, and selected bacterial taxa were quantified by quantitative PCR (qPCR) using group/taxon-specific primers. The parameters observed were the changes in mite population growth and nutritional status, i.e., the total glycogen, lipid, saccharide, and protein contents in mites. The effect of diet perturbation on the variability of the microbiome composition and population growth was lower than the effect induced by mite population. In contrast, the diet perturbation showed a greater effect on nutritional status of mites than the mite population. The endosymbionts exhibited high variations among
T. putrescentiae
populations, including
Cardinium
in the laboratory population,
Blattabacterium
-like bacteria in the dog population, and
Wolbachia
in the dog and Phillips populations.
Solitalea
-like and
Bartonella
-like bacteria were present in the dog, Koppert, and Phillips populations in different proportions. The
T. putrescentiae
microbiome is dynamic and varies based on both the mite population and perturbation; however, the mites remain characterized by robust bacterial communities. Bacterial endosymbionts were found in all populations but represented a dominant portion of the microbiome in only some populations.

IMPORTANCE
We addressed the question of whether population origin or perturbation exerts a more significant influence on the bacterial community of the stored product mite
Tyrophagus putrescentiae
. The microbiomes of four populations of
T. putrescentiae
insects subjected to diet perturbation were compared. Based on our results, the bacterial community was more affected by the mite population than by diet perturbation. This result can be interpreted as indicating high stability of the putative intracellular symbionts in response to dietary perturbation. The changes in the absolute and relative numbers of
Wolbachia
,
Blattabacterium
-like,
Solitalea
-like, and
Cardinium
bacteria in the
T. putrescentiae
populations can also be caused by neutral processes other than perturbation. When nutritional status is considered, the effect of population appeared less important than the perturbation. We hypothesize that differences in the proportions of the endosymbiotic bacteria result in changes in mite population growth.
A Novel Subfamily Esterase with a Homoserine Transacetylase-like Fold but No Transferase Activity
Authors: Li Ping-Yi , Yao Qiong-Qiong , Wang Peng , Zhang Yi , Li Yue , Zhang Yan-Qi , Hao Jie , Zhou Bai-Cheng , Chen Xiu-Lan , Shi Mei , Zhang Yu-Zhong , Zhang Xi-Ying
Published: February 2017

Microbial esterases play important roles in deep-sea organic carbon degradation and cycling. Although they have similar catalytic triads and oxyanion holes, esterases are hydrolases and homoserine transacetylases (HTAs) are transferases. Because two HTA homologs were identified as acetyl esterases, the HTA family was recently divided into the
bona fide
acetyltransferase subfamily and the acetyl esterase subfamily. Here, we identified and characterized a novel HTA-like esterase, Est22, from a deep-sea sedimentary metagenomic library. Est22 could efficiently hydrolyze esters with acyl lengths of up to six carbon atoms but had no transacetylase activity, which is different from HTAs and HTA-like acetyl esterases. Phylogenetic analysis also showed that Est22 and its homologs form a separate branch of the HTA family. We solved the structures of Est22 and its L374D mutant and modeled the structure of the L374D mutant with
p
-nitrophenyl butyrate. Based on structural, mutational, and biochemical analyses, Phe
71
and Met
176
in the oxyanion hole and Arg
294
were revealed to be the key substrate-binding residues. A detailed structural comparison indicated that differences in their catalytic tunnels lead to the different substrate specificities of Est22 and the other two HTA subfamilies. Biochemical and sequence analyses suggested that Est22 homologs may have the same substrate recognition and catalysis mechanisms as Est22. Due to the significant differences in sequences, structures, and substrate specificities between Est22 (and its homologs) and the other two HTA subfamilies, we suggest that Est22 and its homologs represent a new subfamily in the HTA family.

IMPORTANCE
Microbial esterases play important roles in the turnover of organic carbon in the deep sea. Esterases and HTAs represent two groups of α/β hydrolases. Esterases catalyze the hydrolysis of simple esters and are widely used in the pharmaceutical and agrochemical industries, while HTAs catalyze the transfer of an acetyl group from acetyl-coenzyme A (CoA) to homoserine and are essential for microbial growth. Here, we report on a novel HTA-like esterase, Est22, from a deep-sea sediment. Because of the significant differences in sequences, structures, and substrate specificities of HTAs and HTA-like acetyl esterases, Est22 and its homologs represent a new subfamily in the HTA family. This study offers new knowledge regarding marine esterases.
Botrytisfragariae, a New Species Causing Gray Mold on Strawberries, Shows High Frequencies of Specific and Efflux-Based Fungicide Resistance
Authors: Rupp Sabrina , Plesken Cecilia , Rumsey Sibylle , Dowling Madeline , Schnabel Guido , Weber Roland W. S. , Hahn Matthias
Published: February 2017

Botrytis cinerea
causes pre- and postharvest decay of many fruit and vegetable crops. A survey of German strawberry fields revealed
Botrytis
strains that differed from
B. cinerea
in diagnostic PCR markers and growth appearance. Phylogenetic analyses showed that these strains belong to an undescribed species in
Botrytis
clade 2, named
Botrytis
fragariae
sp. nov. Isolates of
B
.
fragariae
were detected in strawberry fields throughout Germany, sometimes at frequencies similar to those of
B. cinerea
, and in the southeastern United States.
B
.
fragariae
was isolated from overwintering strawberry tissue but not from freshly infected fruit.
B
.
fragariae
invaded strawberry tissues with an efficiency similar to or lower than that of
B. cinerea
but showed poor colonization of inoculated nonhost plant tissues. These data and the exclusive occurrence of this fungus on strawberry plants indicate that
B
.
fragariae
is host specific and has a tissue preference different from that of
B. cinerea
. Various fungicide resistance patterns were observed in
B
.
fragariae
populations. Many
B
.
fragariae
strains showed resistance to one or several chemical classes of fungicides and an efflux-based multidrug resistance (MDR1) phenotype previously described in
B. cinerea
. Resistance-related mutations in
B
.
fragariae
were identical or similar to those of
B. cinerea
for carbendazim (E198A mutation in
tubA
), azoxystrobin (G143A in
cytB
), iprodione (G367A+V368F in
bos1
), and MDR1 (gain-of-function mutations in the transcription factor
mrr1
gene and overexpression of the drug efflux transporter gene
atrB
). The widespread occurrence of
B
.
fragariae
indicates that this species is adapted to fungicide-treated strawberry fields and may be of local importance as a gray mold pathogen alongside
B. cinerea
.

IMPORTANCE
Gray mold is the most important fruit rot on strawberries worldwide and requires fungicide treatments for control. For a long time, it was believed to be caused only by
Botrytis cinerea
, a ubiquitous pathogen with a broad host range that quickly develops fungicide resistance. We report the discovery and description of a new species, named
Botrytis
fragariae
, that is widely distributed in commercial strawberry fields in Germany and the southeastern United States. It was observed on overwintering tissue but not on freshly infected fruit and seems host specific on the basis of its occurrence and artificial infection tests.
B
.
fragariae
has also developed resistance to several fungicides that is caused by mutations similar to those known in
B. cinerea
, including an efflux-based multidrug resistance. Our data indicate that
B
.
fragariae
could be of practical importance as a strawberry pathogen in some regions where its abundance is similar to that of
B. cinerea
.
Spread from the Sink to the Patient:
In Situ
Study Using Green Fluorescent Protein (GFP)-Expressing Escherichia coli To Model Bacterial Dispersion from Hand-Washing Sink-Trap Reservoirs
Authors: Kotay Shireen , Chai Weidong , Guilford William , Barry Katie , Mathers Amy J.
Published: February 2017

There have been an increasing number of reports implicating
Gammaproteobacteria
as often carrying genes of drug resistance from colonized sink traps to vulnerable hospitalized patients. However, the mechanism of transmission from the wastewater of the sink P-trap to patients remains poorly understood. Herein we report the use of a designated hand-washing sink lab gallery to model dispersion of green fluorescent protein (GFP)-expressing
Escherichia coli
from sink wastewater to the surrounding environment. We found no dispersion of GFP-expressing
E. coli
directly from the P-trap to the sink basin or surrounding countertop with coincident water flow from a faucet. However, when the GFP-expressing
E. coli
cells were allowed to mature in the P-trap under conditions similar to those in a hospital environment, a GFP-expressing
E. coli
-containing putative biofilm extended upward over 7 days to reach the strainer. This subsequently resulted in droplet dispersion to the surrounding areas (<30 in.) during faucet operation. We also demonstrated that P-trap colonization could occur by retrograde transmission along a common pipe. We postulate that the organisms mobilize up to the strainer from the P-trap, resulting in droplet dispersion rather than dispersion directly from the P-trap. This work helps to further define the mode of transmission of bacteria from a P-trap reservoir to a vulnerable hospitalized patient.

IMPORTANCE
Many recent reports demonstrate that sink drain pipes become colonized with highly consequential multidrug-resistant bacteria, which then results in hospital-acquired infections. However, the mechanism of dispersal of bacteria from the sink to patients has not been fully elucidated. Through establishment of a unique sink gallery, this work found that a staged mode of transmission involving biofilm growth from the lower pipe to the sink strainer and subsequent splatter to the bowl and surrounding area occurs rather than splatter directly from the water in the lower pipe. We have also demonstrated that bacterial transmission can occur via connections in wastewater plumbing to neighboring sinks. This work helps to more clearly define the mechanism and risk of transmission from a wastewater source to hospitalized patients in a world with increasingly antibiotic-resistant bacteria that can thrive in wastewater environments and cause infections in vulnerable patients.
Comparison of Biochemical Properties of the Original and Newly Identified Oleate Hydratases from Stenotrophomonas maltophilia
Authors: Kang Woo-Ri , Seo Min-Ju , Shin Kyung-Chul , Park Jin-Byung , Oh Deok-Kun
Published: February 2017

Oleate hydratases (OhyAs) catalyze the conversion of unsaturated fatty acids to 10-hydroxy fatty acids, which are used as precursors of important industrial compounds, including lactones and ω-hydroxycarboxylic and α,ω-dicarboxylic acids. The genes encoding OhyA and a putative fatty acid hydratase in
Stenotrophomonas maltophilia
were identified by genomic analysis. The putative fatty acid hydratase was purified and identified as an oleate hydratase (OhyA2) based on its substrate specificity. The activity of OhyA2 as a holoenzyme was not affected by adding cofactors, whereas the activity of the original oleate hydratase (OhyA1) showed an increase. Thus, all characterized OhyAs were categorized as either OhyA1 or OhyA2 based on the activities of holoenzymes upon adding cofactors, which were determined by the type of the fourth conserved amino acid of flavin adenine dinucleotide (FAD)-binding motif. The hydration activities of
S. maltophilia
OhyA2 toward unsaturated fatty acids, including oleic acid, palmitoleic acid, linoleic acid, α-linolenic acid, and γ-linolenic acid, were greater than those of OhyA1. Moreover, the specific activity of
S. maltophilia
OhyA2 toward unsaturated fatty acids, with the exception of γ-linolenic acid, was the highest among all reported OhyAs.

IMPORTANCE
All characterized OhyAs were categorized as OhyA1s or OhyA2s based on the different properties of the reported and newly identified holo-OhyAs in
S. maltophilia
upon the addition of cofactors. OhyA2s showed higher activities toward polyunsaturated fatty acids (PUFAs), including linoleic acid, α-linolenic acid, and γ-linolenic acid, than those of OhyA1s. This suggests that OhyA2s can be used more effectively to convert plant oils to 10-hydroxy fatty acids because plant oils contain not only oleic acid but also PUFAs. The hydration activity of the newly identified OhyA2 from
S. maltophilia
toward oleic acid was the highest among the activity levels reported so far. Therefore, this enzyme is an efficient biocatalyst for the conversion of plant oils to 10-hydroxy fatty acids, which can be further converted to important industrial materials.
High-Resolution X-Ray Structures of Two Functionally Distinct Members of the Cyclic Amide Hydrolase Family of Toblerone Fold Enzymes
Authors: Peat Thomas S. , Balotra Sahil , Wilding Matthew , Hartley Carol J. , Newman Janet , Scott Colin
Published: February 2017

The Toblerone fold was discovered recently when the first structure of the cyclic amide hydrolase, AtzD (a cyanuric acid hydrolase), was elucidated. We surveyed the cyclic amide hydrolase family, finding a strong correlation between phylogenetic distribution and specificity for either cyanuric acid or barbituric acid. One of six classes (IV) could not be tested due to a lack of expression of the proteins from it, and another class (V) had neither cyanuric acid nor barbituric acid hydrolase activity. High-resolution X-ray structures were obtained for a class VI barbituric acid hydrolase (1.7 Å) from a
Rhodococcus
species and a class V cyclic amide hydrolase (2.4 Å) from a
Frankia
species for which we were unable to identify a substrate. Both structures were homologous with the tetrameric Toblerone fold enzyme AtzD, demonstrating a high degree of structural conservation within the cyclic amide hydrolase family. The barbituric acid hydrolase structure did not contain zinc, in contrast with early reports of zinc-dependent activity for this enzyme. Instead, each barbituric acid hydrolase monomer contained either Na
+
or Mg
2+
, analogous to the structural metal found in cyanuric acid hydrolase. The
Frankia
cyclic amide hydrolase contained no metal but instead formed unusual, reversible, intermolecular vicinal disulfide bonds that contributed to the thermal stability of the protein. The active sites were largely conserved between the three enzymes, differing at six positions, which likely determine substrate specificity.

IMPORTANCE
The Toblerone fold enzymes catalyze an unusual ring-opening hydrolysis with cyclic amide substrates. A survey of these enzymes shows that there is a good correlation between physiological function and phylogenetic distribution within this family of enzymes and provide insights into the evolutionary relationships between the cyanuric acid and barbituric acid hydrolases. This family of enzymes is structurally and mechanistically distinct from other enzyme families; however, to date the structure of just two, physiologically identical, enzymes from this family has been described. We present two new structures: a barbituric acid hydrolase and an enzyme of unknown function. These structures confirm that members of the CyAH family have the unusual Toblerone fold, albeit with some significant differences.
Could kinesiology taping help mitigate pain, breathlessness and abdominal-related symptoms in cancer?
Authors: Banerjee Gourav , Rose Alison , Briggs Michelle , Johnson Mark I
Published: February 2017

We present the case of a woman who was an amateur athlete diagnosed with primary breast cancer, and 10 years later with terminal metastatic cancer. This case report was prepared posthumously in co-operation with her next of kin (husband). The patient first presented to a sports physiotherapist (AR) for her pain-management and to help maintain physical fitness so that she could continue with sports and an active lifestyle. The patient continued with physiotherapy for several months to enable her to be active. However, when her health deteriorated significantly due to advancing cancer, the treatment was modified and aimed at improving the patient's general well-being. The physiotherapist applied kinesiology tape over the patient's lower rib cage, diaphragm and abdomen in an attempt to manage pain, breathlessness and abdominal bloating. The patient reported alleviation of pain, breathlessness, abdominal discomfort and nausea, accompanied by improvements in eating, drinking, energy levels and physical function.
Rare association of congenital penile urethrocutaneous fistula with Y-type urethral duplication
Authors: Gupta Ashok Kumar , Kumar Manoj , Singh Kawaljit , Sokhal Ashok Kumar
Published: February 2017

A 24-year-old man presented with symptoms of difficulty in passage of urine, mixed with gas and stool from penoscrotal junction since birth. He had a history of surgery (posterior sagittal anorectoplasty) for imperforate anus in first week of his life. On physical examination, external meatus was adequate with small 5×5 mm fistulous opening noted at penoscrotal junction with normal anal tone and opening. On evaluation with retrograde urethrography, voiding cystourethrogram, cystoscopy and barium enema, he was found to have penile urethrocutaneous fistula of 0.5×0.5 cm with Y-type incomplete urethral duplication. Urethroanal fistula (Y-type urethral duplication) was of small size (<5 mm), so closed primarily through perineal route with interposition of ischiorectal fat. Penile urethrocutaneous fistula (4 mm) too was closed primarily and covered with tunica vaginalis flap in single sitting. It has been 6 months since surgery and the patient is fully continent and voiding well with no urinary or faecal leak.