bluuebunny/crossref_metadata_2025_split huggingface dataset:
New Mouse Model for Chronic Infections by Gram-Negative Bacteria Enabling the Study of Anti-Infective Efficacy and Host-Microbe Interactions
Authors: Pletzer Daniel , Mansour Sarah C. , Wuerth Kelli , Rahanjam Negin , Hancock Robert E. W. 
Published: March 2017

ABSTRACT

Only a few, relatively cumbersome animal models enable long-term Gram-negative bacterial infections that mimic human situations, where untreated infections can last for weeks. Here, we describe a simple murine cutaneous abscess model that enables chronic or progressive infections, depending on the subcutaneously injected bacterial strain. In this model,
Pseudomonas aeruginosa
cystic fibrosis epidemic isolate LESB58 caused localized high-density skin and soft tissue infections and necrotic skin lesions for up to 10 days but did not disseminate in either CD-1 or C57BL/6 mice. The model was adapted for use with four major Gram-negative nosocomial pathogens,
Acinetobacter baumannii
,
Klebsiella pneumoniae
,
Enterobacter cloacae
, and
Escherichia coli
. This model enabled noninvasive imaging and tracking of
lux
-tagged bacteria, the influx of activated neutrophils, and production of reactive oxygen-nitrogen species at the infection site. Screening antimicrobials against high-density infections showed that local but not intravenous administration of gentamicin, ciprofloxacin, and meropenem significantly but incompletely reduced bacterial counts and superficial tissue dermonecrosis. Bacterial RNA isolated from the abscess tissue revealed that
Pseudomonas
genes involved in iron uptake, toxin production, surface lipopolysaccharide regulation, adherence, and lipase production were highly upregulated whereas phenazine production and expression of global activator
gacA
were downregulated. The model was validated for studying virulence using mutants of more-virulent
P. aeruginosa
strain PA14. Thus, mutants defective in flagella or motility, type III secretion, or siderophore biosynthesis were noninvasive and suppressed dermal necrosis in mice, while a strain with a mutation in the
bfiS
gene encoding a sensor kinase showed enhanced invasiveness and mortality in mice compared to controls infected with wild-type
P. aeruginosa
PA14.

IMPORTANCE
More than two-thirds of hospital infections are chronic or high-density biofilm infections and difficult to treat due to adaptive, multidrug resistance. Unfortunately, current models of chronic infection are technically challenging and difficult to track without sacrificing animals. Here we describe a model of chronic subcutaneous infection and abscess formation by medically important nosocomial Gram-negative pathogens that is simple and can be used for tracking infections by imaging, examining pathology and immune responses, testing antimicrobial treatments suitable for high-density bacterial infections, and studying virulence. We propose that this mouse model can be a game changer for modeling hard-to-treat Gram-negative bacterial chronic and skin infections.
Toxoplasma DJ-1 Regulates Organelle Secretion by a Direct Interaction with Calcium-Dependent Protein Kinase 1
Authors: Child Matthew A. , Garland Megan , Foe Ian , Madzelan Peter , Treeck Moritz , van der Linden Wouter A. , Oresic Bender Kristina , Weerapana Eranthie , Wilson Mark A. , Boothroyd John C. , Reese Michael L. , Bogyo Matthew 
Published: March 2017

ABSTRACT

Human DJ-1 is a highly conserved and yet functionally enigmatic protein associated with a heritable form of Parkinson’s disease. It has been suggested to be a redox-dependent regulatory scaffold, binding to proteins to modulate their function. Here we present the X-ray crystal structure of the
Toxoplasma
orthologue
Toxoplasma gondii
DJ-1 (TgDJ-1) at 2.1-Å resolution and show that it directly associates with calcium-dependent protein kinase 1 (CDPK1). The TgDJ-1 structure identifies an orthologously conserved arginine dyad that acts as a phospho-gatekeeper motif to control complex formation. We determined that the binding of TgDJ-1 to CDPK1 is sensitive to oxidation and calcium, and that this interaction potentiates CDPK1 kinase activity. Finally, we show that genetic deletion of TgDJ-1 results in upregulation of CDPK1 expression and that disruption of the CDPK1/TgDJ-1 complex
in vivo
prevents normal exocytosis of parasite virulence-associated organelles called micronemes. Overall, our data suggest that TgDJ-1 functions as a noncanonical kinase-regulatory scaffold that integrates multiple intracellular signals to tune microneme exocytosis in
T. gondii
.

IMPORTANCE
Apicomplexan parasites such as
Toxoplasma
and
Plasmodium
are obligate intracellular parasites that require the protective environment of a host cell in order to replicate and survive within a host organism. These parasites secrete effector proteins from specialized apical organelles to select and invade a chosen host cell. The secretion of these organelles is a tightly regulated process coordinated by endogenous small molecules and calcium-dependent protein kinases. We previously identified the
Toxoplasma
orthologue of the highly conserved protein DJ-1 as a regulator of microneme secretion, but the molecular basis for this was not known. We have now identified the molecular mechanism for how TgDJ-1 regulates microneme secretion. TgDJ-1 interacts with the kinase responsible for the secretion of these organelles (calcium-dependent kinase 1) and synergizes with calcium to potentiate kinase activity. This interaction is direct, phosphodependent, and necessary for the normal secretion of these important organelles.
Elevated Cholesterol in the
Coxiella burnetii
Intracellular Niche Is Bacteriolytic
Authors: Mulye Minal , Samanta Dhritiman , Winfree Seth , Heinzen Robert A. , Gilk Stacey D. 
Published: March 2017

ABSTRACT

Coxiella burnetii
is an intracellular bacterial pathogen and a significant cause of culture-negative endocarditis in the United States. Upon infection, the nascent
Coxiella
phagosome fuses with the host endocytic pathway to form a large lysosome-like vacuole called the parasitophorous vacuole (PV). The PV membrane is rich in sterols, and drugs perturbing host cell cholesterol homeostasis inhibit PV formation and bacterial growth. Using cholesterol supplementation of a cholesterol-free cell model system, we found smaller PVs and reduced
Coxiella
growth as cellular cholesterol concentration increased. Further, we observed in cells with cholesterol a significant number of nonfusogenic PVs that contained degraded bacteria, a phenotype not observed in cholesterol-free cells. Cholesterol had no effect on axenic
Coxiella
cultures, indicating that only intracellular bacteria are sensitive to cholesterol. Live-cell microscopy revealed that both plasma membrane-derived cholesterol and the exogenous cholesterol carrier protein low-density lipoprotein (LDL) traffic to the PV. To test the possibility that increasing PV cholesterol levels affects bacterial survival, infected cells were treated with U18666A, a drug that traps cholesterol in lysosomes and PVs. U18666A treatment led to PVs containing degraded bacteria and a significant loss in bacterial viability. The PV pH was significantly more acidic in cells with cholesterol or cells treated with U18666A, and the vacuolar ATPase inhibitor bafilomycin blocked cholesterol-induced PV acidification and bacterial death. Additionally, treatment of infected HeLa cells with several FDA-approved cholesterol-altering drugs led to a loss of bacterial viability, a phenotype also rescued by bafilomycin. Collectively, these data suggest that increasing PV cholesterol further acidifies the PV, leading to
Coxiella
death.

IMPORTANCE
The intracellular Gram-negative bacterium
Coxiella burnetii
is a significant cause of culture-negative infectious endocarditis, which can be fatal if untreated. The existing treatment strategy requires prolonged antibiotic treatment, with a 10-year mortality rate of 19% in treated patients. Therefore, new clinical therapies are needed and can be achieved by better understanding
C. burnetii
pathogenesis. Upon infection of host cells,
C. burnetii
grows within a specialized replication niche, the parasitophorous vacuole (PV). Recent data have linked cholesterol to intracellular
C. burnetii
growth and PV formation, leading us to further decipher the role of cholesterol during
C. burnetii
-host interaction. We observed that increasing PV cholesterol concentration leads to increased acidification of the PV and bacterial death. Further, treatment with FDA-approved drugs that alter host cholesterol homeostasis also killed
C. burnetii
through PV acidification. Our findings suggest that targeting host cholesterol metabolism might prove clinically efficacious in controlling
C. burnetii
infection.
Return to play criteria after hamstring muscle injury in professional football: a Delphi consensus study
Authors: Zambaldi Mattia , Beasley Ian , Rushton Alison 
Published: March 2017

BackgroundHamstring muscle injury (HMI) is the most common injury in professional football and has a high re-injury rate. Despite this, there are no validated criteria to support return to play (RTP) decisions.AimTo use the Delphi method to reach expert consensus on RTP criteria after HMI in professional football.MethodsAll professional football clubs in England (n=92) were invited to participate in a 3-round Delphi study. Round 1 requested a list of criteria used for RTP decisions after HMI. Responses were independently collated by 2 researchers under univocal definitions of RTP criteria. In round 2 participants rated their agreement for each RTP criterion on a 1–5 Likert Scale. In round 3 participants re-rated the criteria that had reached consensus in round 2. Descriptive statistics and Kendall's coefficient of concordance enabled interpretation of consensus.ResultsParticipation rate was limited at 21.7% (n=20), while retention rate was high throughout the 3 rounds (90.0%, 85.0%, 90.0%). Round 1 identified 108 entries with varying definitions that were collated into a list of 14 RTP criteria. Rounds 2 and 3 identified 13 and 12 criteria reaching consensus, respectively. Five domains of RTP assessment were identified: functional performance, strength, flexibility, pain and player's confidence. The highest-rated criteria were in the functional performance domain, with particular importance given to sprint ability.ConclusionThis study defined a list of consensually agreed RTP criteria for HMI in professional football. Further work is now required to determine the validity of the identified criteria.
Cohort profiles of the cross-sectional and prospective participant groups in the second Diabetes MILES—Australia (MILES-2) study
Authors: Browne Jessica L , Holmes-Truscott Elizabeth , Ventura Adriana D , Hendrieckx Christel , Pouwer Frans , Speight Jane 
Published: March 2017

PurposeMore research into the psychosocial aspects of diabetes is needed so that the health and quality of life of people with the condition can be improved. To fill this gap, we conducted the second Diabetes MILES—Australia study (MILES-2), a survey focused on psychological, behavioural and social aspects of diabetes. The aim of the MILES-2 study was to provide a (1) longitudinal follow-up of the original MILES 2011 study cohort; (2) cross-sectional assessment of a new cohort.ParticipantsEligible participants were English-speaking Australians with type 1 or type 2 diabetes, aged 18–75 years. Longitudinal cohort participants were mailed/emailed study invitations directly by researchers. Random sampling (stratified by diabetes type, insulin use, state) of the National Diabetes Services Scheme (NDSS) database and nationwide advertisements were used to recruit new cohort participants. The final sample included N=2342 eligible respondents (longitudinal cohort: n=504; 2015 new cohort: n=1838); 54% had type 2 diabetes.Findings to dateSurvey respondents were from an advantaged socioeconomic background compared to the general population. Respondents with type 1 diabetes were over-represented in the new cohort (45%) relative to the planned stratification (40% type 1 diabetes, 60% type 2 diabetes). Respondents with insulin-treated type 2 diabetes were under-represented in the new cohort relative to the stratified sampling (42% invited vs 50% response). Participants who completed both the 2011 and 2015 surveys were more likely than those completing the 2011 survey only to have type 1 diabetes, report a higher education and annual income, and live in metropolitan areas. Participant feedback indicated that the survey was perceived as relevant and valuable.Future plansThe depth and breadth of the data available in this large sample will highlight unmet needs and priority areas for future investigation and, crucially, will inform policy, programme and intervention development and evaluation in Australia.
Association between ischaemic bowel syndromes and androgen deprivation therapy in patients with prostate cancer: a retrospective cohort study
Authors: Chiang I-Ni , Huang Chao-Yuan , Pu Yeong-Shiau , Chang Chao-Hsiang , Muo Chih-Hsin , Chung Chi-Jung , Wang Ruey-Yun , Young Tai-Horng 
Published: March 2017

ObjectiveThis study investigated the risk of ischaemic bowel syndrome (IBS) in androgen deprivation therapy (ADT) users to explore the long-term outcomes of patients with prostate cancer (PC) receiving ADT treatment.MethodsWe performed a population-based retrospective cohort study. All the clinical information of the study participants were acquired from the Longitudinal Health Insurance Database for Catastrophic Illness Patients in Taiwan. We extracted data for all the patients newly diagnosed with prostate malignancy (ICD-9-CM 185 or C61 in ICD-10-CM) from 2000 to 2008. The patients were then divided into two groups: 7160 male ADT cohort receiving ADT and 7160 male non-ADT comparison group frequency matched by age and index year of ADT treatment of the ADT group. Cox proportional hazard regression was used to estimate the adjusted HR and 95% CIs of the IBS risk.ResultsNo significant difference was noted in the overall incidence rate for IBS between the ADT and non-ADT cohorts (0.86 and 0.89 per 1000 person–year, respectively, p=0.89). Even after adjusting for potential risk factors, a 1.06-fold risk of IBS (95% CI 0.62 to 1.82, p=0.82) was observed in the ADT cohort relative to the non-ADT cohorts. Moreover, we stratified the ADT cohort by time point of ADT treatment after PC diagnosis. Different IBS incidence rates were observed among the early ADT, late-ADT and non-ADT users at 0.77, 1.23 and 0.89 per 1000 person-years, respectively; nonetheless, the difference was not statistically significant. Moreover, no difference was found between the ADT treatment types and IBS risk, including sole orchiectomy, sole luteinising-hormone-releasing hormone and both.ConclusionsResults showed that ADT treatment in patients with PC is not an independent factor for IBS incidence. Large sample sizes for patients with IBS with patients with PC who had received ADT treatment are needed for further study.
Witnessing intimate partner violence and child maltreatment in Ugandan children: a cross-sectional survey
Authors: Devries Karen M , Knight Louise , Child Jennifer C , Kyegombe Nambusi , Hossain Mazeda , Lees Shelley , Watts Charlotte , Naker Dipak 
Published: March 2017

ObjectivesExisting evidence, mainly from high-income countries, shows children who witness intimate partner violence (IPV) at home are more likely to experience other forms of violence, but very little evidence is available from lower income countries. In this paper we aim to explore whether Ugandan children who witness IPV at home are also more likely to experience other forms of maltreatment, factors associated with witnessing and experiencing violence, and whether any increased risk comes from parents, or others outside the home.DesignA representative cross-sectional survey of primary schools.Participants3427 non-boarding primary school students, aged about 11–14 years.SettingLuwero District, Uganda, 2012.MeasuresExposure to child maltreatment was measured using the International Society for the Prevention of Child Abuse and Neglect Child Abuse Screening Tool-Child Institutional, and 2 questions measured witnessing IPV.Results26% of children reported witnessing IPV, but nearly all of these children had also experienced violence themselves. Only 0.6% of boys and 1.6% of girls had witnessed partner violence and not experienced violence. Increased risk of violence was from parents and also from other perpetrators besides parents. Both girls and boys who witnessed and experienced violence had between 1.66 (95% CI 0.96 to 2.87) and 4.50 (95% CI 1.78 to 11.33) times the odds of reporting mental health difficulties, and 3.23 (95% CI 1.99 to 5.24) and 8.12 (95% CI 5.15 to 12.80) times the odds of using physical or sexual violence themselves.ConclusionsIn this sample, witnessing IPV almost never occurred in isolation—almost all children who witnessed partner violence also experienced violence themselves. Our results imply that children in Uganda who are exposed to multiple forms of violence may benefit from intervention to mitigate mental health consequences and reduce use of violence. IPV prevention interventions should be considered to reduce child maltreatment. Large numbers of children also experience maltreatment in homes with no partner violence, highlighting the need for interventions to prevent child maltreatment more broadly.Trial registration numberNCT01678846, results.
Vasopressor use following traumatic injury: protocol for a systematic review
Authors: Hylands Mathieu , Toma Augustin , Beaudoin Nicolas , Frenette Anne-Julie , D'Aragon Frederick , Belley-Côté Emilie , Hylander Morten , Lauzier François , Siemieniuk Reed Alexander , Charbonney Emmanuel , Kwong Joey , Laake Jon Henrik , Guyatt Gordon , Vandvik Per Olav , Rochwerg Bram , Green Robert , Ball Ian , Scales Damon , Murthy Srinivas , Rizoli Sandro , Asfar Pierre , Lamontagne François 
Published: March 2017

IntroductionWorldwide, traumatic casualties are projected to exceed 8 million by year 2020. Haemorrhagic shock and brain injury are the leading causes of death following trauma. While intravenous fluids have traditionally been used to support organ perfusion in the setting of haemorrhage, recent investigations have suggested that restricting fluid therapy by tolerating more severe hypotension may improve survival. However, the safety of permissive hypotension remains uncertain, particularly among patients who have suffered a traumatic brain injury. Vasopressors preferentially vasoconstrict blood vessels that supply non-vital organs and capacitance vessels, thereby mobilising the unstressed blood volume. Used as fluid-sparing adjuncts, these drugs can complement resuscitative measures by correcting hypotension without diluting clotting factors or increasing the risk for tissue oedema.Methods and analysisWe will identify randomised control trials comparing early resuscitation with vasopressors versus placebo or standard care in adults following traumatic injury. Data sources will include MEDLINE, EMBASE, CENTRAL, clinical trial registries and conference proceedings. Two reviewers will independently determine trial eligibility. For each included trial, we will conduct duplicate independent data extraction and risk of bias assessment. We will assess the overall quality of the data for each individual outcome using the GRADE approach.Ethics and disseminationWe will report this review in accordance with the PRISMA statement. We will disseminate our findings at critical care and trauma conferences and through a publication in a peer-reviewed journal. We will also use this systematic review to create clinical guidelines (http://www.magicapp.org), which will be disseminated in a standalone publication.Trial registration numberCRD42016033437.
Parents' views of including young boys in the Swedish national school-based HPV vaccination programme: a qualitative study
Authors: Gottvall Maria , Stenhammar Christina , Grandahl Maria 
Published: March 2017

ObjectiveTo explore parents' views of extending the human papillomavirus (HPV) vaccination programme to also include boys.DesignExplorative qualitative design using individual, face-to-face, interviews and inductive thematic analysis.Setting11 strategically chosen municipalities in central Sweden.ParticipantsParents (n=42) who were offered HPV vaccination for their 11–12 years old daughter in the national school-based vaccination programme.ResultsThe key themes were:equality from a public health perspectiveandperception of risk for disease. Parents expressed low knowledge and awareness about the health benefits of male HPV vaccination, and they perceived low risk for boys to get HPV. Some parents could not see any reason for vaccinating boys. However, many parents preferred gender-neutral vaccination, and some of the parents who had not accepted HPV vaccination for their daughter expressed that they would be willing to accept vaccination for their son, if it was offered. It was evident that there was both trust and distrust in authorities' decision to only vaccinate girls. Parents expressed a preference for increased sexual and reproductive health promotion such as more information about condom use. Some parents shared that it was more important to vaccinate girls than boys since they believed girls face a higher risk of deadly diseases associated with HPV, but some also believed girls might be more vulnerable to side effects of the vaccine.ConclusionsA vaccine offered only to girls may cause parents to be hesitant to vaccinate, while also including boys in the national vaccination programme might improve parents' trust in the vaccine. More information about the health benefits of HPV vaccination for males is necessary to increase HPV vaccination among boys. This may eventually lead to increased HPV vaccine coverage among both girls and boys.
Symptoms and quality of life in patients with suspected angina undergoing CT coronary angiography: a randomised controlled trial
Authors: Williams Michelle C , Hunter Amanda , Shah Anoop , Assi Valentina , Lewis Stephanie , Mangion Kenneth , Berry Colin , Boon Nicholas A , Clark Elizabeth , Flather Marcus , Forbes John , McLean Scott , Roditi Giles , van Beek Edwin JR , Timmis Adam D , Newby David E 
Published: March 2017

BackgroundIn patients with suspected angina pectoris, CT coronary angiography (CTCA) clarifies the diagnosis, directs appropriate investigations and therapies, and reduces clinical events. The effect on patient symptoms is currently unknown.MethodsIn a prospective open-label parallel group multicentre randomised controlled trial, 4146 patients with suspected angina due to coronary heart disease were randomised 1:1 to receive standard care or standard care plus CTCA. Symptoms and quality of life were assessed over 6 months using the Seattle Angina Questionnaire and Short Form 12.ResultsBaseline scores indicated mild physical limitation (74±0.4), moderate angina stability (44±0.4), modest angina frequency (68±0.4), excellent treatment satisfaction (92±0.2) and moderate impairment of quality of life (55±0.3). Compared with standard care alone, CTCA was associated with less marked improvements in physical limitation (difference −1.74 (95% CIs, −3.34 to −0.14), p=0.0329), angina frequency (difference −1.55 (−2.85 to −0.25), p=0.0198) and quality of life (difference −3.48 (−4.95 to −2.01), p<0.0001) at 6 months. For patients undergoing CTCA, improvements in symptoms were greatest in those diagnosed with normal coronary arteries or who had their preventative therapy discontinued, and least in those with moderate non-obstructive disease or had a new prescription of preventative therapy (p<0.001 for all).ConclusionsWhile improving diagnosis, treatment and outcome, CTCA is associated with a small attenuation of the improvements in symptoms and quality of life due to the detection of moderate non-obstructive coronary artery disease.Trial registration number:NCT01149590.