bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative Analysis of the Antimicrobial Activities of Plant Defensin-Like and Ultrashort Peptides against Food-Spoiling Bacteria
Authors: Kraszewska Joanna , Beckett Michael C. , James Tharappel C. , Bond Ursula
Published: May 2016

Antimicrobial peptides offer potential as novel therapeutics to combat food spoilage and poisoning caused by pathogenic and nonpathogenic bacteria. Our previous studies identified the peptide human beta-defensin 3 (HBD3) as a potent antimicrobial agent against a wide range of beer-spoiling bacteria. Thus, HBD3 is an excellent candidate for development as an additive to prevent food and beverage spoilage. To expand the repertoire of peptides with antimicrobial activity against bacteria associated with food spoilage and/or food poisoning, we carried out an
in silico
discovery pipeline to identify peptides with structure and activity similar to those of HBD3, focusing on peptides of plant origin. Using a standardized assay, we compared the antimicrobial activities of nine defensin-like plant peptides to the activity of HBD3. Only two of the peptides, fabatin-2 and Cp-thionin-2, displayed antimicrobial activity; however, the peptides differed from HBD3 in being sensitive to salt and were thermostable. We also compared the activities of several ultrashort peptides to that of HBD3. One of the peptides, the synthetic tetrapeptide O3TR, displayed biphasic antimicrobial activity but had a narrower host range than HBD3. Finally, to determine if the peptides might act in concert to improve antimicrobial activity, we compared the activities of the peptides in pairwise combinations. The plant defensin-like peptides fabatin-2 and Cp-thionin-2 displayed a synergistic effect with HBD3, while O3TR was antagonistic. Thus, some plant defensin-like peptides are effective antimicrobials and may act in concert with HBD3 to control bacteria associated with food spoilage and food poisoning.

IMPORTANCE
Food spoilage and food poisoning caused by bacteria can have major health and economic implications for human society. With the rise in resistance to conventional antibiotics, there is a need to identify new antimicrobials to combat these outbreaks in our food supply. Here we screened plant peptide databases to identify peptides that share structural similarity with the human defensin peptide HBD3, which has known antimicrobial activity against food-spoiling bacteria. We show that two of the plant peptides display antimicrobial activity against bacteria associated with food spoilage. When combined with HBD3, the peptides are highly effective. We also analyzed the activity of an easily made ultrashort synthetic peptide, O3TR. We show that this small peptide also displays antimicrobial activity against food-spoiling bacteria but is not as effective as HBD3 or the plant peptides. The plant peptides identified are good candidates for development as natural additives to prevent food spoilage.
The Cooperative and Interdependent Roles of GerA, GerK, and Ynd in Germination of Bacillus licheniformis Spores
Authors: Borch-Pedersen Kristina , Lindbäck Toril , Madslien Elisabeth H. , Kidd Shani W. , O'Sullivan Kristin , Granum Per Einar , Aspholm Marina
Published: May 2016

When nutrients are scarce,
Bacillus
species form metabolically dormant and extremely resistant spores that enable survival over long periods of time under conditions not permitting growth. The presence of specific nutrients triggers spore germination through interaction with germinant receptors located in the spore's inner membrane.
Bacillus licheniformis
is a biotechnologically important species, but it is also associated with food spoilage and food-borne disease. The
B. licheniformis
ATCC 14580/DSM13 genome exhibits three
gerA
family operons (
gerA
,
gerK
, and
ynd
) encoding germinant receptors. We show that spores of
B. licheniformis
germinate efficiently in response to a range of different single
l
-amino acid germinants, in addition to a weak germination response seen with
d
-glucose. Mutational analyses revealed that the GerA and Ynd germination receptors function cooperatively in triggering an efficient germination response with single
l
-amino acid germinants, whereas the GerK germination receptor is essential for germination with
d
-glucose. Mutant spores expressing only GerA and GerK or only Ynd and GerK show reduced or severely impaired germination responses, respectively, with single
l
-amino acid germinants. Neither GerA nor Ynd could function alone in stimulating spore germination. Together, these results functionally characterize the germination receptor operons present in
B. licheniformis
. We demonstrate the overlapping germinant recognition patterns of the GerA and Ynd germination receptors and the cooperative functionalities between GerA, Ynd, and GerK in inducing germination.

IMPORTANCE
To ensure safe food production and durable foods, there is an obvious need for more knowledge on spore-forming bacteria. It is the process of spore germination that ultimately leads to food spoilage and food poisoning.
Bacillus licheniformis
is a biotechnologically important species that is also associated with food spoilage and food-borne disease. Despite its importance, the mechanisms of spore germination are poorly characterized in this species. This study provides novel knowledge on germination of
B. licheniformis
spores. We characterize the germinant recognition profiles of the three germinant receptors present in
B. licheniformis
spores and demonstrate that the GerA germinant receptor cooperates with the Ynd and GerK germinant receptors to enable an effective germination response to
l
-amino acids. We also demonstrate that GerK is required for germination in response to the single germinant glucose. This study demonstrates the complex interactions between germinant receptors necessary for efficient germination of
B. licheniformis
spores.
Quantitative Distribution of Infectious F-Specific RNA Phage Genotypes in Surface Waters
Authors: Hata Akihiko , Hanamoto Seiya , Shirasaka Yuya , Yamashita Naoyuki , Tanaka Hiroaki
Published: May 2016

F-specific RNA phages (FRNAPHs) are considered potential viral indicators of water pollution due to their occurrence and stability in water environments. However, their suitability as viral indicators is not fully elucidated because the characteristics of FRNAPHs are variable depending on the genotype. In this study, for the characterization of infectious FRNAPH genotypes, integrated culture reverse transcription-PCR coupled with the most probable number approach was applied to surface water samples. Further, to recover low concentrations of FRNAPH genotypes, an FRNAPH recovery method was developed. The novel FRNAPH recovery method using a noncharged microfiltration membrane could effectively recover FRNAPH strains without inactivation, while a method using an electronegative microfiltration membrane resulted in the inactivation of some strains. Infectious FRNAPH genotypes in surface water samples were successfully quantified with an efficiency comparable to that of the conventional plaque assay. Genotype I (GI) and GII FRNAPHs tended to be predominant at locations impacted by treated and untreated municipal wastewater, respectively. The numbers and proportions of infectious FRNAPHs tended to be higher during the winter season when water temperature decreased.

IMPORTANCE
Properties of FRNAPHs are highly variable depending on their genotypes. Previous typing methods for FRNAPHs are not quantitative and/or are based on molecular assays, which cannot differentiate infective strains from inactive strains. Due to the reasons mentioned above, the utility of FRNAPHs as viral indicators of water pollution has not been fully validated. In this study, a quantitative genotyping method for infectious FRNAPHs was developed and applied to surface water samples. The method enabled characterization of infectious FRNAPH genotypes in terms of their occurrence and seasonality. Moreover, comparison of the method to a conventional molecular assay (reverse transcription-quantitative PCR) enabled characterization of their stability. Our approach can provide novel findings for further validation of FRNAPHs as viral indicators of water pollution.
In Situ
Hydrogen Dynamics in a Hot Spring Microbial Mat during a Diel Cycle
Authors: Revsbech Niels Peter , Trampe Erik , Lichtenberg Mads , Ward David M. , Kühl Michael
Published: May 2016

Microbes can produce molecular hydrogen (H
2
) via fermentation, dinitrogen fixation, or direct photolysis, yet the H
2
dynamics in cyanobacterial communities has only been explored in a few natural systems and mostly in the laboratory. In this study, we investigated the diel
in situ
H
2
dynamics in a hot spring microbial mat, where various ecotypes of unicellular cyanobacteria (
Synechococcus
sp.) are the only oxygenic phototrophs. In the evening, H
2
accumulated rapidly after the onset of darkness, reaching peak values of up to 30 μmol H
2
liter
−1
at about 1-mm depth below the mat surface, slowly decreasing to about 11 μmol H
2
liter
−1
just before sunrise. Another pulse of H
2
production, reaching a peak concentration of 46 μmol H
2
liter
−1
, was found in the early morning under dim light conditions too low to induce accumulation of O
2
in the mat. The light stimulation of H
2
accumulation indicated that nitrogenase activity was an important source of H
2
during the morning. This is in accordance with earlier findings of a distinct early morning peak in N
2
fixation and expression of
Synechococcus
nitrogenase genes in mat samples from the same location. Fermentation might have contributed to the formation of H
2
during the night, where accumulation of other fermentation products lowered the pH in the mat to less than pH 6 compared to a spring source pH of 8.3.

IMPORTANCE
Hydrogen is a key intermediate in anaerobic metabolism, and with the development of a sulfide-insensitive microsensor for H
2
, it is now possible to study the microdistribution of H
2
in stratified microbial communities such as the photosynthetic microbial mat investigated here. The ability to measure H
2
profiles within the mat compared to previous measurements of H
2
emission gives much more detailed information about the sources and sinks of H
2
in such communities, and it was demonstrated that the high rates of H
2
formation in the early morning when the mat was exposed to low light intensities might be explained by nitrogen fixation, where H
2
is formed as a by-product.
Regulation of Polyhydroxybutyrate Synthesis in the Soil Bacterium Bradyrhizobium diazoefficiens
Authors: Quelas J. I. , Mesa S. , Mongiardini E. J. , Jendrossek D. , Lodeiro A. R.
Published: May 2016

Polyhydroxybutyrate (PHB) is a carbon and energy reserve polymer in various prokaryotic species. We determined that, when grown with mannitol as the sole carbon source,
Bradyrhizobium diazoefficiens
produces a homopolymer composed only of 3-hydroxybutyrate units (PHB). Conditions of oxygen limitation (such as microoxia, oxic stationary phase, and bacteroids inside legume nodules) were permissive for the synthesis of PHB, which was observed as cytoplasmic granules. To study the regulation of PHB synthesis, we generated mutations in the regulator gene
phaR
and the phasin genes
phaP1
and
phaP4
. Under permissive conditions, mutation of
phaR
impaired PHB accumulation, and a
phaP1 phaP4
double mutant produced more PHB than the wild type, which was accumulated in a single, large cytoplasmic granule. Moreover, PhaR negatively regulated the expression of
phaP1
and
phaP4
as well as the expression of
phaA1
and
phaA2
(encoding a 3-ketoacyl coenzyme A [CoA] thiolases),
phaC1
and
phaC2
(encoding PHB synthases), and
fixK
2
(encoding a cyclic AMP receptor protein [CRP]/fumarate and nitrate reductase regulator [FNR]-type transcription factor of genes for microoxic lifestyle). In addition to the depressed PHB cycling,
phaR
mutants accumulated more extracellular polysaccharides and promoted higher plant shoot dry weight and competitiveness for nodulation than the wild type, in contrast to the
phaC1
mutant strain, which is defective in PHB synthesis. These results suggest that
phaR
not only regulates PHB granule formation by controlling the expression of phasins and biosynthetic enzymes but also acts as a global regulator of excess carbon allocation and symbiosis by controlling
fixK
2
.

IMPORTANCE
In this work, we investigated the regulation of polyhydroxybutyrate synthesis in the soybean-nodulating bacterium
Bradyrhizobium diazoefficiens
and its influence in bacterial free-living and symbiotic lifestyles. We uncovered a new interplay between the synthesis of this carbon reserve polymer and the network responsible for microoxic metabolism through the interaction between the gene regulators
phaR
and
fixK
2
. These results contribute to the understanding of the physiological conditions required for polyhydroxybutyrate biosynthesis. The interaction between these two main metabolic pathways is also reflected in the symbiotic phenotypes of soybeans inoculated with
phaR
mutants, which were more competitive for nodulation and enhanced dry matter production by the plants. Therefore, this knowledge may be applied to the development of superior strains to be used as improved inoculants for soybean crops.
Educational differences in sickness absence trends among young employees from 2002 to 2013 in Helsinki, Finland
Authors: Sumanen Hilla , Lahelma Eero , Lahti Jouni , Pietiläinen Olli , Rahkonen Ossi
Published: May 2016

Objective

Socioeconomic differences in sickness absence (SA) are well established among older employees but poorly understood among the young. Our aim was to examine 12-year trends in educational differences in SA among young female and male employees, and to assess the magnitude of the differences.

Design

We examined annual SA spells. The data were obtained from the employer's registers and linked to Statistics Finland's register data on completed education and qualifications. Education was classified into four hierarchical groups. Joinpoint regression models were used to identify turning points in SA trends. The magnitude of the relative educational differences was estimated in accordance with the relative index of inequality for 2002, 2008 and 2013.

Setting

Employees of the City of Helsinki, Finland, in 2002–2013.

Participants

The analyses covered female and male employees aged 25–34 years: employees aged 35–54 years were used as a reference group.

Outcome

SA spells.

Results

An educational gradient emerged among younger and older women and men. SA spells increased in the early 2000s, and downward turning points were located in 2007–2010 in all educational groups among women and in most groups among men. The magnitude of the differences remained broadly stable among younger women from 2002 to 2013, and decreased slightly among older women and more strongly among younger and older men. The educational differences were greater among men than women in the early 2000s, but similar among both at the end of the study period.

Conclusions

The changes in SA spells may reflect the economic downturn started in 2008 and resulting job insecurity. Early preventive measures aimed at reducing educational differences in SA should be focused at an early stage on those with low levels of education in particular.
Does a strategy to promote shared decision-making reduce medical practice variation in the choice of either single or double embryo transfer after in vitro fertilisation? A secondary analysis of a randomised controlled trial
Authors: Brabers Anne E M , van Dijk Liset , Groenewegen Peter P , van Peperstraten Arno M , de Jong Judith D
Published: May 2016

Objectives

The hypothesis that shared decision-making (SDM) reduces medical practice variations is increasingly common, but no evidence is available. We aimed to elaborate further on this, and to perform a first exploratory analysis to examine this hypothesis. This analysis, based on a limited data set, examined how SDM is associated with variation in the choice of single embryo transfer (SET) or double embryo transfer (DET) after in vitro fertilisation (IVF). We examined variation between and within hospitals.

Design

A secondary analysis of a randomised controlled trial.

Setting

5 hospitals in the Netherlands.

Participants

222 couples (woman aged <40 years) on a waiting list for a first IVF cycle, who could choose between SET and DET (ie, ≥2 embryos available).

Intervention

SDM via a multifaceted strategy aimed to empower couples in deciding how many embryos should be transferred. The strategy consisted of decision aid, support of IVF nurse and the offer of reimbursement for an extra treatment cycle. Control group received standard IVF care.

Outcome measure

Difference in variation due to SDM in the choice of SET or DET, both between and within hospitals.

Results

There was large variation in the choice of SET or DET between hospitals in the control group. Lower variation between hospitals was observed in the group with SDM. Within most hospitals, variation in the choice of SET or DET appeared to increase due to SDM. Variation particularly increased in hospitals where mainly DET was chosen in the control group.

Conclusions

Although based on a limited data set, our study gives a first insight that including patients’ preferences through SDM results in less variation between hospitals, and indicates another pattern of variation within hospitals. Variation that results from patient preferences could be potentially named the informed patient rate. Our results provide the starting point for further research.

Trial registration number

NCT00315029; Post-results.
From QASC to QASCIP: successful Australian translational scale-up and spread of a proven intervention in acute stroke using a prospective pre-test/post-test study design
Authors: Middleton Sandy , Lydtin Anna , Comerford Daniel , Cadilhac Dominique A , McElduff Patrick , Dale Simeon , Hill Kelvin , Longworth Mark , Ward Jeanette , Cheung N Wah , D'Este Cate
Published: May 2016

Objectives

To embed an evidence-based intervention to manageFEver, hyperglycaemia (Sugar) andSwallowing (the FeSS protocols) in stroke, previously demonstrated in the Quality in Acute Stroke Care (QASC) trial to decrease 90-day death and dependency, into all stroke services in New South Wales (NSW), Australia's most populous state.

Design

Pre-test/post-test prospective study.

Setting

36 NSW stroke services.

Methods

Our clinical translational initiative, the QASC Implementation Project (QASCIP), targeted stroke services to embed 3 nurse-led clinical protocols (the FeSS protocols) into routine practice. Clinical champions attended a 1-day multidisciplinary training workshop and received standardised educational resources and ongoing support. Using the National Stroke Foundation audit collection tool and processes, patient data from retrospective medical record self-reported audits for 40 consecutive patients with stroke per site pre-QASCIP (1 July 2012 to 31 December 2012) were compared with prospective self-reported data from 40 consecutive patients with stroke per site post-QASCIP (1 November 2013 to 28 February 2014). Inter-rater reliability was substantial for 10 of 12 variables.

Primary outcome measures

Proportion of patients receiving care according to the FeSS protocols pre-QASCIP to post-QASCIP.

Results

All 36 (100%) NSW stroke services participated, nominating 100 site champions who attended our educational workshops. The time from start of intervention to completion of post-QASCIP data collection was 8 months. All (n=36, 100%) sites provided medical record audit data for 2144 patients (n=1062 pre-QASCIP; n=1082 post-QASCIP). Pre-QASCIP to post-QASCIP, proportions of patients receiving the 3 targeted clinical behaviours increased significantly: management of fever (pre: 69%; post: 78%; p=0.003), hyperglycaemia (pre: 23%; post: 34%; p=0.0085) and swallowing (pre: 42%; post: 51%; p=0.033).

Conclusions

We obtained unprecedented statewide scale-up and spread to all NSW stroke services of a nurse-led intervention previously proven to improve long-term patient outcomes. As clinical leaders search for strategies to improve quality of care, our initiative is replicable and feasible in other acute care settings.
Opportunities to improve clinical summaries for patients at hospital discharge
Authors: Sarzynski Erin , Hashmi Hamza , Subramanian Jeevarathna , Fitzpatrick Laurie , Polverento Molly , Simmons Michael , Brooks Kevin , Given Charles
Published: May 2016

Background

Clinical summaries are electronic health record (EHR)-generated documents given to hospitalised patients during the discharge process to review their hospital stays and inform postdischarge care. Presently, it is unclear whether clinical summaries include relevant content or whether healthcare organisations configure their EHRs to generate content in a way that promotes patient self-management after hospital discharge. We assessed clinical summaries in three relevant domains: (1) content; (2) organisation; and (3) readability, understandability and actionability.

Methods

Two authors performed independent retrospective chart reviews of 100 clinical summaries generated at two Michigan hospitals using different EHR vendors for patients discharged 1 April –30 June 2014. We developed an audit tool based on the Meaningful Use view-download-transmit objective and the Society of Hospital Medicine Discharge Checklist (content); the Institute of Medicine recommendations for distributing easy-to-understand print material (organisation); and five readability formulas and the Patient Education Materials Assessment Tool (readability, understandability and actionability).

Results

Clinical summaries averaged six pages (range 3–12). Several content elements were universally auto-populated into clinical summaries (eg, medication lists); others were not (eg, care team). Eighty-five per cent of clinical summaries contained discharge instructions, more often generated from third-party sources than manually entered by clinicians. Clinical summaries contained an average of 14 unique messages, including non-clinical elements irrelevant to postdischarge care. Medication list organisation reflected reconciliation mandates, and dosing charts, when present, did not carry column headings over to subsequent pages. Summaries were written at the 8th–12th grade reading level and scored poorly on assessments of understandability and actionability. Inter-rater reliability was strong for most elements in our audit tool.

Conclusions

Our study highlights opportunities to improve clinical summaries for guiding patients' postdischarge care.
Artemisinins as potential anticancer agents: uptake detection in erythrocytes using Fourier transform infrared spectroscopy and cytotoxicity against bladder cancer cells
Authors: Humphreys Charlotte , Cooper Alan J , Barbu Eugen , Birch Brian R , Lwaleed Bashir A
Published: May 2016

Aims

Semisynthetic derivatives of the antimalarial drug artemisinin may also possess anticancer properties. The ability to detect artemisinin uptake and distribution in cells would facilitate live cell imaging without labelling. This study describes mid-range infrared absorption spectra for three artemisinin variants and attempts to detect their presence in a simple cell model (erythrocytes). Cytotoxicity assays assess potential anticancer properties against bladder cancer cells.

Methods

Mid-range Fourier transform infrared spectra were obtained from dry preparations of dihydroartemisinin (DHA), artesunate (ART) and artemether (ARTE). Erythrocytes were prepared from normal blood and incubated for 30 min at 37°C with the three artemisinin derivatives. Cytospin preparations were prepared on aluminium foil for spectroscopy. Potential for growth inhibition in the RT112 bladder carcinoma cell line was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide residual viable biomass method.

Results

Spectra were obtained from the three native compounds. Repeat scans after 8 weeks showed ART and ARTE to be stable, stored under manufacturer's recommendations. DHA exhibited marked changes over the same period. It was possible by subtraction to detect DHA in cytospins, but not ART or ARTE. The fit between the subtraction spectrum and that of the native compound was >80%. DHA and ART showed strong cytotoxic potential against RT112 cells.

Conclusions

The artemisinin derivatives tested exhibit unique mid-range infrared absorption spectra which can be used to monitor degradation and, for DHA, can be detected by subtraction in loaded erythrocytes rendering future imaging studies feasible. Its cytotoxic efficacy against RT112 cells suggests bladder cancer as a possible target disease.
North Atlantic humpback whale (Megaptera novaeangliae) hotspots defined by bathymetric features off western Puerto Rico
Authors: MacKay M.M. , Würsig B. , Bacon C.E. , Selwyn J.D.
Published: May 2016

North Atlantic humpback whales (Megaptera novaeangliae (Borowski, 1781)) are increasing in number, necessitating current data from winter areas for assessing potential interactions with humans. Occurrence patterns of humpback whales wintering off Puerto Rico were investigated to predict where whales aggregate in nearshore areas. Here we describe the relationship between group associations of humpback whales and bathymetric features off western Puerto Rico. Data were collected from 2011 to 2014. Effort consisted of 240.9 vessel h, 13.0 aerial h, and 303.6 h of land observations conducted over 165 days. A total of 197 humpback whale groups were observed with n = 331 individuals: 91 (46.2%) singletons, 67 (34%) dyads, 17 (8.6%) mother–calf pairs, 8 (4.1%) competitive groups, 8 (4.1%) mother–calf–escort groups, and 6 (3.1%) mixed-species associations. A linear regression model supported that group composition correlated with hotspots associated with four bathymetric features. Dyads and competitive groups were dispersed among features in deeper water. Singletons were observed farther from a shelf edge, whereas singing males were closely associated with a shelf edge. Mother–calf pairs occurred nearshore in shallow water; however, when mother–calf pairs were sighted with an escort, they were offshore. This study is especially important ahead of possible removal from the Endangered Species list.
Distinct right ventricle remodeling in response to pressure overload in the rat
Authors: Mendes-Ferreira P. , Santos-Ribeiro D. , Adão R. , Maia-Rocha C. , Mendes-Ferreira M. , Sousa-Mendes C. , Leite-Moreira A. F. , Brás-Silva C.
Published: May 2016

Pulmonary arterial hypertension (PAH), the most serious chronic disorder of the pulmonary circulation, is characterized by pulmonary vasoconstriction and remodeling, resulting in increased afterload on the right ventricle (RV). In fact, RV function is the main determinant of prognosis in PAH. The most frequently used experimental models of PAH include monocrotaline- and chronic hypoxia-induced PAH, which primarily affect the pulmonary circulation. Alternatively, pulmonary artery banding (PAB) can be performed to achieve RV overload without affecting the pulmonary vasculature, allowing researchers to determine the RV-specific effects of their drugs/interventions. In this work, using two different degrees of pulmonary artery constriction, we characterize, in full detail, PAB-induced adaptive and maladaptive remodeling of the RV at 3 wk after PAB surgery. Our results show that application of a mild constriction resulted in adaptive hypertrophy of the RV, with preserved systolic and diastolic function, while application of a severe constriction resulted in maladaptive hypertrophy, with chamber dilation and systolic and diastolic dysfunction up to the isolated cardiomyocyte level. By applying two different degrees of constriction, we describe, for the first time, a reliable and short-duration PAB model in which RV adaptation can be distinguished at 3 wk after surgery. We characterize, in full detail, structural and functional changes of the RV in its response to moderate and severe constriction, allowing researchers to better study RV physiology and transition to dysfunction and failure, as well as to determine the effects of new therapies.
A myosin activator improves actin assembly and sarcomere function of human-induced pluripotent stem cell-derived cardiomyocytes with a troponin T point mutation
Authors: Broughton K. M. , Li J. , Sarmah E. , Warren C. M. , Lin Y.-H. , Henze M. P. , Sanchez-Freire V. , Solaro R. J. , Russell B.
Published: May 2016

We have investigated cardiac myocytes derived from human-induced pluripotent stem cells (iPSC-CMs) from two normal control and two family members expressing a mutant cardiac troponin T (cTnT-R173W) linked to dilated cardiomyopathy (DCM). cTnT is a regulatory protein of the sarcomeric thin filament. The loss of this basic charge, which is strategically located to control tension, has consequences leading to progressive DCM. iPSC-CMs serve as a valuable platform for understanding clinically relevant mutations in sarcomeric proteins; however, there are important questions to be addressed with regard to myocyte adaptation that we model here by plating iPSC-CMs on softer substrates (100 kPa) to create a more physiologic environment during recovery and maturation of iPSC-CMs after thawing from cryopreservation. During the first week of culture of the iPSC-CMs, we have determined structural and functional characteristics as well as actin assembly dynamics. Shortening, actin content, and actin assembly dynamics were depressed in CMs from the severely affected mutant at 1 wk of culture, but by 2 wk differences were less apparent. Sarcomeric troponin and myosin isoform composition were fetal/neonatal. Furthermore, the troponin complex, reconstituted with wild-type cTnT or recombinant cTnT-R173W, depressed the entry of cross-bridges into the force-generating state, which can be reversed by the myosin activator omecamtiv mecarbil. Therapeutic doses of this drug increased both contractility and the content of F-actin in the mutant iPSC-CMs. Collectively, our data suggest the use of a myosin activation reagent to restore function within patient-specific iPSC-CMs may aid in understanding and treating this familial DCM.
Matrix metalloproteinase-2 in oncostatin M-induced sarcomere degeneration in cardiomyocytes
Authors: Fan Xiaohu , Hughes Bryan G. , Ali Mohammad A. M. , Chan Brandon Y. H. , Launier Katherine , Schulz Richard
Published: May 2016

Cardiomyocyte dedifferentiation may be an important source of proliferating cardiomyocytes facilitating cardiac repair. Cardiomyocyte dedifferentiation and proliferation induced by oncostatin-M (OSM) is characterized by sarcomere degeneration. However, the mechanism underlying sarcomere degeneration remains unclear. We hypothesized that this process may involve matrix metalloproteinase-2 (MMP-2), a key protease localized at the sarcomere in cardiomyocytes. We tested the hypothesis that MMP-2 is involved in the sarcomere degeneration that characterizes cardiomyocyte dedifferentiation. Confocal immunofluorescence and biochemical methods were used to explore the role of MMP-2 in OSM-induced dedifferentiation of neonatal rat ventricular myocytes (NRVM). OSM caused a concentration- and time-dependent loss of sarcomeric α-actinin and troponin-I in NRVM. Upon OSM-treatment, the mature sarcomere transformed to a phenotype resembling a less-developed sarcomere, i.e., loss of sarcomeric proteins and Z-disk transformed into disconnected Z bodies, characteristic of immature myofibrils. OSM dose dependently increased MMP-2 activity. Both the pan-MMP inhibitor GM6001 and the selective MMP-2 inhibitor ARP 100 prevented sarcomere degeneration induced by OSM treatment. OSM also induced NRVM cell cycling and increased methyl-thiazolyl-tetrazolium (MTT) staining, preventable by MMP inhibition. These results suggest that MMP-2 mediates sarcomere degeneration in OSM-induced cardiomyocyte dedifferentiation and thus potentially contributes to cardiomyocyte regeneration.
Discovery and identification of candidate sex-related genes based on transcriptome sequencing of Russian sturgeon (Acipenser gueldenstaedtii) gonads
Authors: Chen Yadong , Xia Yongtao , Shao Changwei , Han Lei , Chen Xuejie , Yu Mengjun , Sha Zhenxia
Published: May 2016

As the Russian sturgeon ( Acipenser gueldenstaedtii) is an important food and is the main source of caviar, it is necessary to discover the genes associated with its sex differentiation. However, the complicated life and maturity cycles of the Russian sturgeon restrict the accurate identification of sex in early development. To generate a first look at specific sex-related genes, we sequenced the transcriptome of gonads in different development stages (1, 2, and 5 yr old stages) with next-generation RNA sequencing. We generated >60 million raw reads, and the filtered reads were assembled into 263,341 contigs, which produced 38,505 unigenes. Genes involved in signal transduction mechanisms were the most abundant, suggesting that development of sturgeon gonads is under control of signal transduction mechanisms. Differentially expressed gene analysis suggests that more genes for protein synthesis, cytochrome c oxidase subunits, and ribosomal proteins were expressed in female gonads than in male. Meanwhile, male gonads expressed more transposable element transposase, reverse transcriptase, and transposase-related genes than female. In total, 342, 782, and 7,845 genes were detected in intersex, male, and female transcriptomes, respectively. The female gonad expressed more genes than the male gonad, and more genes were involved in female gonadal development. Genes (sox9, foxl2) are differentially expressed in different sexes and may be important sex-related genes in Russian sturgeon. S ox9 genes are responsible for the development of male gonads and foxl2 for female gonads.
Chronic Recreational Physical Inactivity and Epithelial Ovarian Cancer Risk: Evidence from the Ovarian Cancer Association Consortium
Authors: Cannioto Rikki , LaMonte Michael J. , Risch Harvey A. , Hong Chi-Chen , Sucheston-Campbell Lara E. , Eng Kevin H. , Brian Szender J. , Chang-Claude Jenny , Schmalfeldt Barbara , Klapdor Ruediger , Gower Emily , Minlikeeva Albina N. , Zirpoli Gary R. , Bandera Elisa V. , Berchuck Andrew , Cramer Daniel , Doherty Jennifer A. , Edwards Robert P. , Fridley Brooke L. , Goode Ellen L. , Goodman Marc T. , Hogdall Estrid , Hosono Satoyo , Jensen Allan , Jordan Susan ,   on behalf of The Australian Ovarian Cancer Study Group, Kjaer Susanne K. , Matsuo Keitaro , Ness Roberta B. , Olsen Catherine M. , Olson Sara H. , Leigh Pearce Celeste , Pike Malcolm C. , Anne Rossing Mary , Szamreta Elizabeth A. , Thompson Pamela J. , Tseng Chiu-Chen , Vierkant Robert A. , Webb Penelope M. , Wentzensen Nicolas , Wicklund Kristine G. , Winham Stacey J. , Wu Anna H. , Modugno Francesmary , Schildkraut Joellen M. , Terry Kathryn L. , Kelemen Linda E. , Moysich Kirsten B.
Published: May 2016

Background: Despite a large body of literature evaluating the association between recreational physical activity and epithelial ovarian cancer (EOC) risk, the extant evidence is inconclusive, and little is known about the independent association between recreational physical inactivity and EOC risk. We conducted a pooled analysis of nine studies from the Ovarian Cancer Association Consortium to investigate the association between chronic recreational physical inactivity and EOC risk.

Methods: In accordance with the 2008 Physical Activity Guidelines for Americans, women reporting no regular, weekly recreational physical activity were classified as inactive. Multivariable logistic regression was utilized to estimate the ORs and 95% confidence intervals (CI) for the association between inactivity and EOC risk overall and by subgroups based upon histotype, menopausal status, race, and body mass index.

Results: The current analysis included data from 8,309 EOC patients and 12,612 controls. We observed a significant positive association between inactivity and EOC risk (OR = 1.34; 95% CI, 1.14–1.57), and similar associations were observed for each histotype.

Conclusions: In this large pooled analysis examining the association between recreational physical inactivity and EOC risk, we observed consistent evidence of an association between chronic inactivity and all EOC histotypes.

Impact: These data add to the growing body of evidence suggesting that inactivity is an independent risk factor for cancer. If the apparent association between inactivity and EOC risk is substantiated, additional work via targeted interventions should be pursued to characterize the dose of activity required to mitigate the risk of this highly fatal disease. Cancer Epidemiol Biomarkers Prev; 25(7); 1114–24. ©2016 AACR.
ASH1L Links Histone H3 Lysine 36 Dimethylation to MLL Leukemia
Authors: Zhu Li , Li Qin , Wong Stephen H.K. , Huang Min , Klein Brianna J. , Shen Jinfeng , Ikenouye Larissa , Onishi Masayuki , Schneidawind Dominik , Buechele Corina , Hansen Loren , Duque-Afonso Jesús , Zhu Fangfang , Martin Gloria Mas , Gozani Or , Majeti Ravindra , Kutateladze Tatiana G. , Cleary Michael L.
Published: May 2016

Numerous studies in multiple systems support that histone H3 lysine 36 dimethylation (H3K36me2) is associated with transcriptional activation; however, the underlying mechanisms are not well defined. Here, we show that the H3K36me2 chromatin mark written by the ASH1L histone methyltransferase is preferentially bound in vivo by LEDGF, a mixed-lineage leukemia (MLL)–associated protein that colocalizes with MLL, ASH1L, and H3K36me2 on chromatin genome wide. Furthermore, ASH1L facilitates recruitment of LEDGF and wild-type MLL proteins to chromatin at key leukemia target genes and is a crucial regulator of MLL-dependent transcription and leukemic transformation. Conversely, KDM2A, an H3K36me2 demethylase and Polycomb group silencing protein, antagonizes MLL-associated leukemogenesis. Our studies are the first to provide a basic mechanistic insight into epigenetic interactions wherein placement, interpretation, and removal of H3K36me2 contribute to the regulation of gene expression and MLL leukemia, and suggest ASH1L as a novel target for therapeutic intervention.

Significance: Epigenetic regulators play vital roles in cancer pathogenesis and represent a new frontier in therapeutic targeting. Our studies provide basic mechanistic insight into the role of H3K36me2 in transcription activation and MLL leukemia pathogenesis and implicate ASH1L histone methyltransferase as a promising target for novel molecular therapy. Cancer Discov; 6(7); 770–83. ©2016 AACR.

See related commentary by Balbach and Orkin, p. 700.

This article is highlighted in the In This Issue feature, p. 681
Formyl-Peptide Receptor 2/3/Lipoxin A
4
Receptor Regulates Neutrophil-Platelet Aggregation and Attenuates Cerebral Inflammation
Authors: Vital Shantel A. , Becker Felix , Holloway Paul M. , Russell Janice , Perretti Mauro , Granger D. Neil , Gavins Felicity N.E.
Published: May 2016

Background—

Platelet activation at sites of vascular injury is essential for hemostasis, but it is also a major pathomechanism underlying ischemic injury. Because anti-inflammatory therapies limit thrombosis and antithrombotic therapies reduce vascular inflammation, we tested the therapeutic potential of 2 proresolving endogenous mediators, annexin A1 N-terminal derived peptide (AnxA1
Ac2–26
) and aspirin-triggered lipoxin A
4
(15-epi-lipoxin A
4
), on the cerebral microcirculation after ischemia/reperfusion injury. Furthermore, we tested whether the lipoxin A
4
receptor formyl-peptide receptor 2/3 (Fpr2/3; ortholog to human FPR2/lipoxin A
4
receptor) evoked neuroprotective functions after cerebral ischemia/reperfusion injury.

Methods and Results—

Using intravital microscopy, we found that cerebral ischemia/reperfusion injury was accompanied by neutrophil and platelet activation and neutrophil-platelet aggregate formation within cerebral microvessels. Moreover, aspirin-triggered lipoxin A
4
activation of neutrophil Fpr2/3 regulated neutrophil-platelet aggregate formation in the brain and inhibited the reactivity of the cerebral microvasculature. The same results were obtained with AnxA1
Ac2–26
administration. Blocking Fpr2/lipoxin A
4
receptor with the antagonist Boc2 reversed this effect, and treatments were ineffective in Fpr2/3 knockout mice, which displayed an exacerbated disease severity, evidenced by increased infarct area, blood-brain barrier dysfunction, increased neurological score, and elevated levels of cytokines. Furthermore, aspirin treatment significantly reduced cerebral leukocyte recruitment and increased endogenous levels of aspirin-triggered lipoxin A
4
, effects again mediated by Fpr2/3.

Conclusion—

Fpr2/lipoxin A
4
receptor is a therapeutic target for initiating endogenous proresolving, anti-inflammatory pathways after cerebral ischemia/reperfusion injury.
Variant Aldehyde Dehydrogenase 2 (
ALDH2*2
) Is a Risk Factor for Coronary Spasm and ST‐Segment Elevation Myocardial Infarction
Authors: Mizuno Yuji , Hokimoto Seiji , Harada Eisaku , Kinoshita Kenji , Nakagawa Kazuko , Yoshimura Michihiro , Ogawa Hisao , Yasue Hirofumi
Published: May 2016

Background

Mitochondrial aldehyde dehydrogenase 2 (
ALDH
2) plays a key role in removing toxic aldehydes. Deficient variant

ALDH
2*2

genotype is prevalent in up to 40% of the East Asians and reported to be associated with acute myocardial infarction (
AMI
). To elucidate the mechanisms underlying the association of

ALDH
2*2

with
AMI
, we compared the clinical features of
AMI
patients with

ALDH
2*2

to those with wild‐type

ALDH
2*1/*1

.

Methods and Results

The study subjects consisted of 202 Japanese patients with acute
ST‐
segment elevation myocardial infarction (
STEMI
) (156 men and 46 women; mean age, 67.3±12.0) who underwent primary percutaneous coronary intervention (
PCI
). In 85 patients, provocation test for coronary spasm was also done 6 month post‐
PCI
.

ALDH
2

genotyping was performed by direct application of the TaqMan polymerase chain system. Of the 202 patients, 103 (51.0%) were carriers of

ALDH
2*2

and 99 (49.0%) those of

ALDH
2*1/*1

. There were no differences in clinical features between

ALDH
2*2

and

ALDH
2*1/*1

carrier groups except higher frequencies of coronary spasm and alcohol flush syndrome (
AFS
) (88.6% vs 56.1%;
P
=0.001 and 94.3% vs 17.6%;
P
<0.001), less‐frequent alcohol habit (14.6% vs 51.5%;
P
<0.001), and higher peak plasma creatine phophokinase levels (2224 vs 1617 mg/
dL
;
P
=0.002) in the

ALDH
2*2

than the

ALDH
2*1/*1

carrier group.

Conclusions

ALDH
2*2

is prevalent (51.0%) among Japanese
STEMI
patients, and those with

ALDH
2*2

had higher frequencies of coronary spasm and
AFS
and more‐severe myocardial injury compared to those with

ALDH
2*1/*1

.
The Group Dynamics of Interorganizational Relationships
Authors: Davis Jason P.
Published: May 2016

This paper examines how organizations collaborate with multiple partners, such as when they develop innovative and complex product platforms like smartphones, servers, and MRI machines that rely on technologies developed by organizations in three or more sectors. Research on multipartner alliances often treats them as a collection of independent dyads, neglecting the possibility of third-party influence and interference in dyads that can inhibit innovation. Using a multiple-case, inductive study of six groups, each composed of three organizations engaged in technology and product development in the computer industry, I examine the collaborative forms and processes that organizations use to innovate with multiple partners in groups. Groups that used the collaborative forms of independent parallel dyads or single unified triads generated mistrust and conflict that stemmed from expectations about third-party participation and overlapping roles and thus had low innovation performance and weaker ties. Other groups avoided these problems by using a dynamic collaboration process that I call “group cycling,” in which managers viewed their triad as a small group, decomposed innovative activities into a series of interlinked dyads between different pairs of partners, and managed third-party interests across time. By temporarily restricting participation to pairs, managers chose which ideas, technologies, and resources to incorporate from third parties into single dyads and ensured that the outputs of multiple dyads were combined into a broader innovative whole.
X-Ray Fluorescence Analysis and Self-Organizing Maps Classification of the Etruscan Gold Coin Collection at the Monetiere of Florence
Authors: Arias Claudio , Bani Stefano , Catalli Fiorenzo , Lorenzetti Giulia , Grifoni Emanuela , Legnaioli Stefano , Pagnotta Stefano , Palleschi Vincenzo
Published: May 2016

The “Monetiere” of Florence hosts the most important collection of Etruscan coins in the world. In the framework of the longstanding collaboration between the Monetiere and the Applied Laser and Spectroscopy Laboratory in Pisa, the Etruscan gold coin collection of the museum was studied. The measurements were performed at the Monetiere, using a portable energy-dispersive X-ray fluorescence (XRF) instrument. The quantitative determination of the gold alloys used for the realization of the coins was obtained applying the fundamental parameters method to the XRF spectra; as a result, using the self-organizing maps method, we were able to classify the coins in four main groups. The main parameter determining the classification is the quantity of silver in the alloy. The results obtained shed some light about the origin of the coins under study.
Repeat the Song! The Psalms and the Eucharist
Authors: Daffern Megan
Published: May 2016

In prayer and ritual, time and eternity intersect. The theology of time is a lens through which such comparable texts may be examined. Time is also a means for reflecting on hermeneutic method in studying prayer texts. This article explores time and eternity in both the prayer texts and the study of the prayer texts of the Psalms and the Eucharist, with reference to the work of Eliade, Levine, and Begbie. The two prayer texts, and the three theologians, thus contribute much to the interpretation of one another when read together.
Premature Triumphalism in Corinth
Authors: Malcolm Matthew R.
Published: May 2016

Challenges to the hypothesis of over realised eschatology behind 1 Corinthians must be taken into account. However, the broad idea that the Corinthians were behaving as though they could lay claim to eschatological arrival is worthy of renewed consideration. This article argues for a modified version of the hypothesis, using the phrase “premature triumphalism”. This conception emphasises the problem as chiefly behavioural rather than doctrinal. Further, the problem is seen to be largely unwitting, being implicit, rather than explicit, in its eschatological claims. It is an issue perceived by Paul, rather than fully by the Corinthians.
Commemorating 500 Years of the Greek New Testament in Print: Erasmus and his Legacy
Authors: Moore Richard K.
Published: May 2016

Desiderius Erasmus was the first to make the Greek New Testament available in printed form. By doing so he established the form the Greek New Testament would take for almost four hundred years. Consequently, vernacular translations based on the Greek reflected that form. In the English-speaking world, where the King James Version dominated until the 1980s, his influence persisted for over 450 years. Some aspects of it persist in the majority of English versions in use today.