bluuebunny/crossref_metadata_2025_split huggingface dataset:
Treatment interruption in HIV‐positive patients followed up in Cameroon's antiretroviral treatment programme: individual and health care supply‐related factors (ANRS‐12288 EVOLCam survey)
Authors: Tong Christelle , Suzan‐Monti Marie , Sagaon‐Teyssier Luis , Mimi Mohamed , Laurent Christian , Maradan Gwenaëlle , Mengue Marie‐Thérèse , Spire Bruno , Kuaban Christopher , Vidal Laurent , Boyer Sylvie ,   the EVOL Cam group
Published: January 2018

SummaryIntroductionDecreasing international financial resources for HIV and increasing numbers of antiretroviral treatment (ART)‐treated patients may jeopardise treatment continuity in low‐income settings. Using data from the EVOLCam ANRS‐12288 survey, this study aimed to document the prevalence of unplanned treatment interruption for more than 2 consecutive days (TI>2d) and investigate the associated individual and health care supply‐related factors within the Cameroonian ART programme.MethodsA cross‐sectional mixed methods survey was carried out between April and December 2014 in 19 HIV services of the Centre and Littoral regions. A multilevel logistic model was estimated on 1885 ART‐treated patients in these services to investigate factors of TI>2d in the past 4 weeks.ResultsAmong the study population, 403 (21%) patients reported TI>2d. Patients followed up in hospitals reporting ART stock‐outs were more likely to report TI>2d while those followed up in the Littoral region, in medium‐ or small‐sized hospitals and in HIV services proposing financial support were at lower risk of TI>2d. The following individual factors were also associated with a lower risk of TI>2d: living in a couple, having children, satisfaction with attention provided by doctor, tuberculosis co‐infection and not having consulted a traditional healer.ConclusionsBesides identifying individual factors of TI>2d, our study highlighted the role of health care supply‐related factors in shaping TI in Cameroon's ART programme, especially the deleterious effect of ART stock‐outs. Our results also suggest that the high proportion of patients reporting TI could jeopardise progress in the fight against HIV in the country, unless effective measures are quickly implemented like ensuring the continuity of ART supply.
Shank3‐deficient thalamocortical neurons show HCN channelopathy and alterations in intrinsic electrical properties
Authors: Zhu Mengye , Idikuda Vinay Kumar , Wang Jianbing , Wei Fusheng , Kumar Virang , Shah Nikhil , Waite Christopher B. , Liu Qinglian , Zhou Lei 
Published: January 2018

Key points
Shank3 increases the HCN channel surface expression in heterologous expression systems.
Shank3Δ13–16 deficiency causes significant reduction in HCN2 expression and Ih current amplitude in thalamocortical (TC) neurons.
Shank3Δ13–16‐ but not Shank3Δ4–9‐deficient TC neurons share changes in basic electrical properties which are comparable to those of HCN2−/− TC neurons.
HCN channelopathy may critically mediate events downstream from Shank3 deficiency.
AbstractSHANK3 is a scaffolding protein that is highly enriched in excitatory synapses. Mutations in the SHANK3 gene have been linked to neuropsychiatric disorders especially the autism spectrum disorders. SHANK3 deficiency is known to cause impairments in synaptic transmission, but its effects on basic neuronal electrical properties that are more localized to the soma and proximal dendrites remain unclear. Here we confirmed that in heterologous expression systems two different mouse Shank3 isoforms, Shank3A and Shank3C, significantly increase the surface expression of the mouse hyperpolarization‐activated, cyclic‐nucleotide‐gated (HCN) channel. In Shank3Δ13–16 knockout mice, which lack exons 13–16 in the Shank3 gene (both Shank3A and Shank3C are removed) and display a severe behavioural phenotype, the expression of HCN2 is reduced to an undetectable level. The thalamocortical (TC) neurons from the ventrobasal (VB) complex of Shank3Δ13–16 mice demonstrate reduced Ih current amplitude and correspondingly increased input resistance, negatively shifted resting membrane potential, and abnormal spike firing in both tonic and burst modes. Impressively, these changes closely resemble those of HCN2−/− TC neurons but not of the TC neurons from Shank3Δ4–9 mice, which lack exons 4–9 in the Shank3 gene (Shank3C still exists) and demonstrate moderate behavioural phenotypes. Additionally, Shank3 deficiency increases the ratio of excitatory/inhibitory balance in VB neurons but has a limited impact on the electrical properties of connected thalamic reticular (RTN) neurons. These results provide new understanding about the role of HCN channelopathy in mediating detrimental effects downstream from Shank3 deficiency.
Germination, Outgrowth, and Vegetative-Growth Kinetics of Dry-Heat-Treated Individual Spores of Bacillus Species
Authors: He Lin , Chen Zhan , Wang Shiwei , Wu Muying , Setlow Peter , Li Yong-qing 
Published: January 2018

ABSTRACT

DNA damage kills dry-heated spores of
Bacillus subtilis
, but dry-heat-treatment effects on spore germination and outgrowth have not been studied. This is important, since if dry-heat-killed spores germinate and undergo outgrowth, toxic proteins could be synthesized. Here, Raman spectroscopy and differential interference contrast microscopy were used to study germination and outgrowth of individual dry-heat-treated
B. subtilis
and
Bacillus megaterium
spores. The major findings in this work were as follows: (i) spores dry-heat-treated at 140°C for 20 min lost nearly all viability but retained their Ca
2+
-dipicolinic acid (CaDPA) depot; (ii) in most cases, dry-heat treatment increased the average times and variability of all major germination events in
B. subtilis
spore germination with nutrient germinants or CaDPA, and in one nutrient germination event with
B. megaterium
spores; (iii)
B. subtilis
spore germination with dodecylamine, which activates the spore CaDPA release channel, was unaffected by dry-heat treatment; (iv) these results indicate that dry-heat treatment likely damages spore proteins important in nutrient germinant recognition and cortex peptidoglycan hydrolysis, but not CaDPA release itself; and (v) analysis of single spores incubated on nutrient-rich agar showed that while dry-heat-treated spores that are dead can complete germination, they cannot proceed into outgrowth and thus not to vegetative growth. The results of this study provide new information on the effects of dry heat on bacterial spores and indicate that dry-heat sterilization regimens should produce spores that cannot outgrow and thus cannot synthesize potentially dangerous proteins.

IMPORTANCE
Much research has shown that high-temperature dry heat is a promising means for the inactivation of spores on medical devices and spacecraft decontamination. Dry heat is known to kill
Bacillus subtilis
spores by DNA damage. However, knowledge about the effects of dry-heat treatment on spore germination and outgrowth is limited, especially at the single spore level. In the current work, Raman spectroscopy and differential interference contrast microscopy were used to analyze CaDPA levels in and kinetics of nutrient- and non-nutrient germination of multiple individual dry-heat-treated
B. subtilis
and
Bacillus megaterium
spores that were largely dead. The outgrowth and subsequent cell division of these germinated but dead dry-heat-treated spores were also examined. The knowledge obtained in this study will help understand the effects of dry heat on spores both on Earth and in space, and indicates that dry heat can be safely used for sterilization purposes.
Parainfluenza Virus Infection Sensitizes Cancer Cells to DNA-Damaging Agents: Implications for Oncolytic Virus Therapy
Authors: Fox Candace R. , Parks Griffith D. 
Published: January 2018

ABSTRACT

A parainfluenza virus 5 (PIV5) with mutations in the P/V gene (P/V-CPI
−
) is restricted for spread in normal cells but not in cancer cells
in vitro
and is effective at reducing tumor burdens in mouse model systems. Here we show that P/V-CPI
−
infection of HEp-2 human laryngeal cancer cells results in the majority of the cells dying, but unexpectedly, over time, there is an emergence of a population of cells that survive as P/V-CPI
−
persistently infected (PI) cells. P/V-CPI
−
PI cells had elevated levels of basal caspase activation, and viability was highly dependent on the activity of cellular inhibitor-of-apoptosis proteins (IAPs) such as Survivin and XIAP. In challenge experiments with external inducers of apoptosis, PI cells were more sensitive to cisplatin-induced DNA damage and cell death. This increased cisplatin sensitivity correlated with defects in DNA damage signaling pathways such as phosphorylation of Chk1 and translocation of damage-specific DNA binding protein 1 (DDB1) to the nucleus. Cisplatin-induced killing of PI cells was sensitive to the inhibition of wild-type (WT) p53-inducible protein 1 (WIP1), a phosphatase which acts to terminate DNA damage signaling pathways. A similar sensitivity to cisplatin was seen with cells during acute infection with P/V-CPI
−
as well as during acute infections with WT PIV5 and the related virus human parainfluenza virus type 2 (hPIV2). Our results have general implications for the design of safer paramyxovirus-based vectors that cannot establish PI as well as the potential for combining chemotherapy with oncolytic RNA virus vectors.

IMPORTANCE
There is intense interest in developing oncolytic viral vectors with increased potency against cancer cells, particularly those cancer cells that have gained resistance to chemotherapies. We have found that infection with cytoplasmically replicating parainfluenza virus can result in increases in the killing of cancer cells by agents that induce DNA damage, and this is linked to alterations to DNA damage signaling pathways that balance cell survival versus death. Our results have general implications for the design of safer paramyxovirus-based vectors that cannot establish persistent infection, the repurposing of drugs that target cellular IAPs as antivirals, and the combined use of DNA-damaging chemotherapy agents in conjunction with oncolytic RNA virus vectors.
Attenuation of Simian Varicella Virus Infection by Enhanced Green Fluorescent Protein in Rhesus Macaques
Authors: Mahalingam Ravi , Kaufer Benedikt B. , Ouwendijk Werner J. D. , Verjans Georges M. G. M. , Coleman Colin , Hunter Meredith , Das Arpita , Palmer Brent E. , Clambey Eric , Nagel Maria A. , Traina-Dorge Vicki 
Published: January 2018

ABSTRACT

Simian varicella virus (SVV), the primate counterpart of varicella-zoster virus, causes varicella (chickenpox), establishes latency in ganglia, and reactivates to produce zoster. We previously demonstrated that a recombinant SVV expressing enhanced green fluorescent protein (rSVV.eGFP) is slightly attenuated both in culture and in infected monkeys. Here, we generated two additional recombinant SVVs to visualize infected cells
in vitro
and
in vivo
. One harbors eGFP fused to the N terminus of open reading frame 9 (ORF9) (rSVV.eGFP-2a-ORF9), and another harbors eGFP fused to the C terminus of ORF66 (rSVV.eGFP-ORF66). Both recombinant viruses efficiently expressed eGFP in cultured cells. Both recombinant SVV infections in culture were comparable to that of wild-type SVV (SVV.wt). Unlike SVV.wt, eGFP-tagged SVV did not replicate in rhesus cells in culture. Intratracheal (i.t.) or i.t. plus intravenous (i.v.) inoculation of rhesus macaques with these new eGFP-tagged viruses resulted in low viremia without varicella rash, although SVV DNA was abundant in bronchoalveolar lavage (BAL) fluid at 10 days postinoculation (dpi). SVV DNA was also found in trigeminal ganglia of one monkey inoculated with rSVV.eGFP-ORF66. Intriguingly, a humoral response to both SVV and eGFP was observed. In addition, monkeys inoculated with the eGFP-expressing viruses were protected from superinfection with SVV.wt, suggesting that the monkeys had mounted an efficient immune response. Together, our results show that eGFP expression could be responsible for their reduced pathogenesis.

IMPORTANCE
SVV infection in nonhuman primates has served as an extremely useful animal model to study varicella-zoster virus (VZV) pathogenesis. eGFP-tagged viruses are a great tool to investigate their pathogenesis. We constructed and tested two new recombinant SVVs with eGFP inserted into two different locations in the SVV genome. Both recombinant SVVs showed robust replication in culture but reduced viremia compared to that with SVV.wt during primary infection in rhesus macaques. Our results indicate that conclusions on eGFP-tagged viruses based on
in vitro
results should be handled with care, since eGFP expression could result in attenuation of the virus.
Inter-rater reliability in the Paediatric Observation Priority Score (POPS)
Authors: Langton Lisa , Bonfield Adam , Roland Damian 
Published: January 2018

ObjectiveThe primary objective of this study was to determine the level of inter-rater reliability between nursing staff for the Paediatric Observation Priority Score (POPS).DesignRetrospective observational study.SettingSingle-centre paediatric emergency department.Participants12 participants from a convenience sample of 21 nursing staff.InterventionsParticipants were shown video footage of three pre-recorded paediatric assessments and asked to record their own POPS for each child. The participants were blinded to the original, in-person POPS. Further data were gathered in the form of a questionnaire to determine the level of training and experience the candidate had using the POPS score prior to undertaking this study.Main outcome measuresInter-rater reliability among participants scoring of the POPS.ResultsOverall kappa value for case 1 was 0.74 (95% CI 0.605 to 0.865), case 2 was 1 (perfect agreement) and case 3 was 0.66 (95% CI 0.58 to 0.744).ConclusionThis study suggests there is good inter-rater reliability between different nurses’ use of POPS in assessing sick children in the emergency department.
What are the sources of stress and distress for general practitioners working in England? A qualitative study
Authors: Riley Ruth , Spiers Johanna , Buszewicz Marta , Taylor Anna Kathryn , Thornton Gail , Chew-Graham Carolyn Anne 
Published: January 2018

ObjectivesThis paper reports the sources of stress and distress experienced by general practitioners (GP) as part of a wider study exploring the barriers and facilitators to help-seeking for mental illness and burnout among this medical population.DesignQualitative study using in-depth interviews with 47 GP participants. The interviews were audio-recorded, transcribed, anonymised and imported into NVivo V.11 to facilitate data management. Data were analysed using a thematic analysis employing the constant comparative method.SettingEngland.ParticipantsA purposive sample of GP participants who self-identified as: (1) currently living with mental distress, (2) returning to work following treatment, (3) off sick or retired early as a result of mental distress or (4) without experience of mental distress. Interviews were conducted face-to-face or over the telephone.ResultsThe key sources of stress/distress related to: (1) emotion work—the work invested and required in managing and responding to the psychosocial component of GPs’ work, and dealing with abusive or confrontational patients; (2) practice culture—practice dynamics and collegial conflict, bullying, isolation and lack of support; (3) work role and demands—fear of making mistakes, complaints and inquests, revalidation, appraisal, inspections and financial worries.ConclusionIn addition to addressing escalating workloads through the provision of increased resources, addressing unhealthy practice cultures is paramount. Collegial support, a willingness to talk about vulnerability and illness, and having open channels of communication enable GPs to feel less isolated and better able to cope with the emotional and clinical demands of their work. Doctors, including GPs, are not invulnerable to the clinical and emotional demands of their work nor the effects of divisive work cultures—culture change and access to informal and formal support is therefore crucial in enabling GPs to do their job effectively and to stay well.
Survival in relation to multimorbidity patterns in older adults in primary care in Barcelona, Spain (2010–2014): a longitudinal study based on electronic health records
Authors: Ibarra-Castillo Carolina , Guisado-Clavero Marina , Violan-Fors Concepció , Pons-Vigués Mariona , López-Jiménez Tomàs , Roso-Llorach Albert 
Published: January 2018

BackgroundSeveral studies have analysed the characteristics of multimorbidity patterns but none have evaluated the relationship with survival. The purpose of this study was to compare survival across older adults with different chronic multimorbidity patterns (CMPs).MethodsProspective longitudinal observational study using electronic health records for 190 108 people aged ≥65 years in Barcelona, Spain (2009–2014). CMPs were identified by cluster analysis. Mortality rates were estimated using the Catalan population structure and individual time at risk. Survival according to CMP (Cox regression) was analysed using hazard ratios (HRs) and 95% confidence intervals (CIs) with stratification by sex and age group (65–79, 80–94) and adjustment for age at onset, deprivation index, number of chronic conditions and invoiced drugs.ResultsThe highest mortality rates were observed in men, adults aged 80–94 years, socially disadvantaged quintiles and people prescribed more drugs and with fewer conditions. Using the musculoskeletal pattern as the reference category, men with the digestive-respiratory pattern had a higher risk of death, with adjusted HRs of 6.16 (95% CI 5.37 to 7.06) in the 65–79 age group and 2.62 (95% CI 2.31 to 2.97) in the 80–94 age group. In women, the cardiovascular pattern was associated with the highest risk, with adjusted HRs of 6.34 (95% CI 5.28 to 7.61) in the 65–79 age group and 3.05 (95% CI 2.73 to 3.41) in the 80–94 age group. These patterns were also associated with the highest mortality rates.ConclusionsMortality and survival vary according to CMPs in older adults stratified by sex and age. Our findings are useful for guiding the design and implementation of clinical management strategies.
Role of germline aberrations affecting CTNNA1, MAP3K6 and MYD88 in gastric cancer susceptibility
Authors: Weren Robbert D A , van der Post Rachel S , Vogelaar Ingrid P , van Krieken J Han , Spruijt Liesbeth , Lubinski Jan , Jakubowska Anna , Teodorczyk Urszula , Aalfs Cora M , van Hest Liselotte P , Oliveira Carla , Kamping Eveline J , Schackert Hans K , Ranzani Guglielmina N , Gómez García Encarna B , Hes Frederik J , Holinski-Feder Elke , Genuardi Maurizio , Ausems Margreet G E M , Sijmons Rolf H , Wagner Anja , van der Kolk Lizet E , Cats Annemieke , Bjørnevoll Inga , Hoogerbrugge Nicoline , Ligtenberg Marjolijn J L 
Published: January 2018

BackgroundIn approximately 10% of all gastric cancer (GC) cases, a heritable cause is suspected. A subset of these cases have a causative germline CDH1 mutation; however, in most cases the cause remains unknown. Our objective was to assess to what extent these remaining cases may be explained by germline mutations in the novel candidate GC predisposing genes CTNNA1, MAP3K6 or MYD88.MethodsWe sequenced a large cohort of unexplained young and/or familial patients with GC (n=286) without a CDH1germline mutation for germline variants affecting CTNNA1, MAP3K6 and MYD88 using a targeted next-generation sequencing approach based on single-molecule molecular inversion probes.ResultsPredicted deleterious germline variants were not encountered in MYD88, but recurrently observed in CTNNA1 (n=2) and MAP3K6 (n=3) in our cohort of patients with GC. In contrast to deleterious variants in CTNNA1, deleterious variants in MAP3K6 also occur frequently in the general population.ConclusionsBased on our results MAP3K6 should no longer be considered a GC predisposition gene, whereas deleterious CTNNA1 variants are confirmed as an infrequent cause of GC susceptibility. Biallelic MYD88 germline mutations are at most a very rare cause of GC susceptibility as no additional cases were identified.
Fabry Disease: prevalence of affected males and heterozygotes with pathogenic GLA mutations identified by screening renal, cardiac and stroke clinics, 1995–2017
Authors: Doheny Dana , Srinivasan Ram , Pagant Silvere , Chen Brenden , Yasuda Makiko , Desnick Robert J 
Published: January 2018

BackgroundFabry Disease (FD), an X linked lysosomal storage disease due to pathogenic α-galactosidase A (GLA) mutations, results in two major subtypes, the early-onset Type 1 ‘Classic’ and the Type 2 ‘Later-Onset’ phenotypes. To identify previously unrecognised patients, investigators screened cardiac, renal and stroke clinics by enzyme assays. However, some screening studies did not perform confirmatory GLA mutation analyses, and many included recently recognised ‘benign/likely-benign’ variants, thereby inflating prevalence estimates.MethodsOnline databases were searched for all FD screening studies in high-risk clinics (1995–2017). Studies reporting GLA mutations were re-analysed for pathogenic mutations, sex and phenotype. Phenotype-specific and sex-specific prevalence rates were determined.ResultsOf 67 studies, 63 that screened 51363patients (33943M and 17420F) and provided GLA mutations were reanalysed for disease-causing mutations. Of reported GLA mutations, benign variants occurred in 47.9% of males and 74.1% of females. The following were the revised prevalence estimates: among 36820 (23954M and 12866F) haemodialysis screenees, 0.21% males and 0.15% females; among 3074 (2031M and 1043F) renal transplant screenees, 0.25% males and no females; among 5491 (4054M and 1437F) cardiac screenees, 0.94% males and 0.90% females; and among 5978 (3904M and 2074F) stroke screenees, 0.13% males and 0.14% females. Among male and female screenees with pathogenic mutations, the type 1 Classic phenotype was predominant (~60%), except more male cardiac patients (75%) had type 2 Later-Onset phenotype.ConclusionsCompared with previous findings, reanalysis of 63 studies increased the screenee numbers (~3.4-fold), eliminated 20 benign/likely benign variants, and provided more accurate sex-specific and phenotype-specific prevalence estimates, ranging from ~0.13% of stroke to ~0.9% of cardiac male or female screenees.