bluuebunny/crossref_metadata_2025 huggingface dataset:
Intraclade Variability in Toxin Production and Cytotoxicity of Bacillus cereus Group Type Strains and Dairy-Associated Isolates
Authors: Miller Rachel A. , Jian Jiahui , Beno Sarah M. , Wiedmann Martin , Kovac Jasna
Published: January 2018

While some species in the
Bacillus cereus
group are well-characterized human pathogens (e.g.,
B. anthracis
and
B. cereus sensu stricto
), the pathogenicity of other species (e.g.,
B. pseudomycoides
) either has not been characterized or is presently not well understood. To provide an updated characterization of the pathogenic potential of species in the
B. cereus
group, we classified a set of 52 isolates, including 8 type strains and 44 isolates from dairy-associated sources, into 7 phylogenetic clades and characterized them for (i) the presence of toxin genes, (ii) phenotypic characteristics used for identification, and (iii) cytotoxicity to human epithelial cells. Overall, we found that
B. cereus
toxin genes are broadly distributed but are not consistently present within individual species and/or clades. After growth at 37°C, isolates within a clade did not typically show a consistent cytotoxicity phenotype, except for isolates in clade VI (
B. weihenstephanensis
/
B. mycoides
), where none of the isolates were cytotoxic, and isolates in clade I (
B. pseudomycoides
), which consistently displayed cytotoxic activity. Importantly, our study highlights that
B. pseudomycoides
is cytotoxic toward human cells. Our results indicate that the detection of toxin genes does not provide a reliable approach to predict the pathogenic potential of
B. cereus
group isolates, as the presence of toxin genes is not always consistent with cytotoxicity phenotype. Overall, our results suggest that isolates from multiple
B. cereus
group clades have the potential to cause foodborne illness, although cytotoxicity is not always consistently found among isolates within each clade.

IMPORTANCE
Despite the importance of the
Bacillus cereus
group as a foodborne pathogen, characterizations of the pathogenic potential of all
B. cereus
group species were lacking. We show here that
B. pseudomycoides
(clade I), which has been considered a harmless environmental microorganism, produces toxins and exhibits a phenotype consistent with the production of pore-forming toxins. Furthermore,
B. mycoides/B. weihenstephanensis
isolates (clade VI) did not show cytotoxicity when grown at 37°C, despite carrying multiple toxin genes. Overall, we show that the current standard methods to characterize
B. cereus
group isolates and to detect the presence of toxin genes are not reliable indicators of species, phylogenetic clades, or an isolate's cytotoxic capacity, suggesting that novel methods are still needed for differentiating pathogenic from nonpathogenic species within the
B. cereus
group. Our results also contribute data that are necessary to facilitate risk assessments and a better understanding as to which
B. cereus
group species are likely to cause foodborne illness.
Both Major Histocompatibility Complex Class I (MHC-I) and MHC-II Molecules Are Required, while MHC-I Appears To Play a Critical Role in Host Defense against Primary Coxiella burnetii Infection
Authors: Buttrum Laura , Ledbetter Lindsey , Cherla Rama , Zhang Yan , Mitchell William J. , Zhang Guoquan
Published: January 2018

To understand the role of class I major histocompatibility complex (MHC-I) and class II MHC (MHC-II) antigen presentation pathways in host defense againstCoxiella burnetiiinfection, we examined whether MHC-I or MHC-II deficiency in mice would significantly influence their susceptibility to virulentC. burnetiiNine Mile phase I (NMI) infection. The results indicate that NMI infection induced more severe disease in both MHC-I-deficient and MHC-II-deficient mice than in wild-type (WT) mice, while only MHC-I-deficient mice developed a severe persistent infection and were unable to control bacterial replication. These results suggest that both MHC-I-restricted CD8+T cells and MHC-II-restricted CD4+T cells contribute to host defense against primaryC. burnetiiinfection, while MHC-I-restricted CD8+T cells appear to play a more critical role in controlling bacterial replication. Additionally, although NMI infection induced more severe disease in TAP1-deficient mice than in their WT counterparts, TAP1 deficiency in mice did not significantly influence their ability to eliminateC. burnetii. This suggests thatC. burnetiiantigen presentation to CD8+T cells by the MHC-I classical pathway may depend only partially on TAP1. Furthermore, granzyme B deficiency in mice did not significantly alter their susceptibility toC. burnetiiinfection, but perforin-deficient mice were unable to control host inflammatory responses during primaryC. burnetiiinfection. These results suggest that perforin, but not granzyme B, is required forC. burnetiiantigen-specific cytotoxic CD8+T cells to control primaryC. burnetiiinfection.
A Multimodal Strategy Used by a Large c-di-GMP Network
Authors: Dahlstrom Kurt M. , Collins Alan J. , Doing Georgia , Taroni Jaclyn N. , Gauvin Timothy J. , Greene Casey S. , Hogan Deborah A. , O'Toole George A.
Published: January 2018

The
Pseudomonas fluorescens
genome encodes more than 50 proteins predicted to be involved in c-di-GMP signaling. Here, we demonstrated that, tested across 188 nutrients, these enzymes and effectors appeared capable of impacting biofilm formation. Transcriptional analysis of network members across ∼50 nutrient conditions indicates that altered gene expression can explain a subset of but not all biofilm formation responses to the nutrients. Additional organization of the network is likely achieved through physical interaction, as determined via probing ∼2,000 interactions by bacterial two-hybrid assays. Our analysis revealed a multimodal regulatory strategy using combinations of ligand-mediated signals, protein-protein interaction, and/or transcriptional regulation to fine-tune c-di-GMP-mediated responses. These results create a profile of a large c-di-GMP network that is used to make important cellular decisions, opening the door to future model building and the ability to engineer this complex circuitry in other bacteria.

IMPORTANCE
Cyclic diguanylate (c-di-GMP) is a key signaling molecule regulating bacterial biofilm formation, and many microbes have up to dozens of proteins that make, break, or bind this dinucleotide. A major open issue in the field is how signaling specificity is conferred in the unpartitioned space of a bacterial cell. Here, we took a systems approach, using mutational analysis, transcriptional studies, and bacterial two-hybrid analysis to interrogate this network. We found that a majority of enzymes are capable of impacting biofilm formation in a context-dependent manner, and we revealed examples of two or more modes of regulation (i.e., transcriptional control with protein-protein interaction) being utilized to generate an observable impact on biofilm formation.
ERK Is a Critical Regulator of JC Polyomavirus Infection
Authors: DuShane Jeanne K. , Wilczek Michael P. , Mayberry Colleen L. , Maginnis Melissa S.
Published: January 2018

The human JC polyomavirus (JCPyV) infects the majority of the population worldwide and presents as an asymptomatic, persistent infection in the kidneys. In individuals who are immunocompromised, JCPyV can become reactivated and cause a lytic infection in the central nervous system resulting in the fatal, demyelinating disease progressive multifocal leukoencephalopathy (PML). Infection is initiated by interactions between the capsid protein viral protein 1 (VP1) and the α2,6-linked sialic acid on lactoseries tetrasaccharide c (LSTc), while JCPyV internalization is facilitated by 5-hydroxytryptamine 2 receptors (5-HT
2
Rs). The mechanisms by which the serotonin receptors mediate virus entry and the signaling cascades required to drive viral infection remain poorly understood. JCPyV was previously shown to induce phosphorylation of extracellular signal-regulated kinase (ERK), a downstream target of the mitogen-activated protein kinase (MAPK) pathway, upon virus entry. However, it remained unclear whether ERK activation was required for JCPyV infection. Both ERK-specific small interfering RNA (siRNA) and ERK inhibitor treatments resulted in significantly diminished JCPyV infection in both kidney and glial cells yet had no effect on the infectivity of the polyomavirus simian virus 40 (SV40). Experiments characterizing the role of ERK during steps in the viral life cycle indicate that ERK activation is required for viral transcription, as demonstrated by a significant reduction in production of large T antigen (TAg), a key viral protein associated with the initiation of viral transcription and viral replication. These findings delineate the role of the MAPK-ERK signaling pathway in JCPyV infection, elucidating how the virus reprograms the host cell to promote viral pathogenesis.

IMPORTANCE
Viral infection is dependent upon host cell factors, including the activation of cellular signaling pathways. These interactions between viruses and host cells are necessary for infection and play an important role in viral disease outcomes. The focus of this study was to determine how the human JC polyomavirus (JCPyV), a virus that resides in the kidney of the majority of the population and can cause the fatal, demyelinating disease progressive multifocal leukoencephalopathy (PML) in the brains of immunosuppressed individuals, usurps a cellular signaling pathway to promote its own infectious life cycle. We demonstrated that the activation of extracellular signal-regulated kinase (ERK), a component of the mitogen-activated protein kinase (MAPK) pathway, promotes JCPyV transcription, which is required for viral infection. Our findings demonstrate that the MAPK-ERK signaling pathway is a key determinant of JCPyV infection, elucidating new information regarding the signal reprogramming of host cells by a pathogenic virus.
HIV-1-Specific IgA Monoclonal Antibodies from an HIV-1 Vaccinee Mediate Galactosylceramide Blocking and Phagocytosis
Authors: Wills Saintedym , Hwang Kwan-Ki , Liu Pinghuang , Dennison S. Moses , Tay Matthew Zirui , Shen Xiaoying , Pollara Justin , Lucas Judith T. , Parks Robert , Rerks-Ngarm Supachai , Pitisuttithum Punnee , Nitayapan Sorachai , Kaewkungwal Jaranit , Thomas Rasmi , Kim Jerome H. , Michael Nelson L. , Robb Merlin L. , McRaven Mike , Montefiori David C. , Hope Thomas J. , Liao Hua-Xin , Moody M. Anthony , Ferrari Guido , Haynes Barton F. , Alam S. Munir , Bonsignori Mattia , Tomaras Georgia D.
Published: January 2018

Vaccine-elicited humoral immune responses comprise an array of antibody forms and specificities, with only a fraction contributing to protective host immunity. Elucidation of antibody effector functions responsible for protective immunity against human immunodeficiency virus type 1 (HIV-1) acquisition is a major goal for the HIV-1 vaccine field. Immunoglobulin A (IgA) is an important part of the host defense against pathogens; however, little is known about the role of vaccine-elicited IgA and its capacity to mediate antiviral functions. To identify the antiviral functions of HIV-1-specific IgA elicited by vaccination, we cloned HIV-1 envelope-specific IgA monoclonal antibodies (MAbs) by memory B cell cultures from peripheral blood mononuclear cells from an RV144 vaccinee and produced two IgA clonal cell lines (HG129 and HG130) producing native, nonrecombinant IgA MAbs. The HG129 and HG130 MAbs mediated phagocytosis by monocytes, and HG129 blocked HIV-1 Env glycoprotein binding to galactosylceramide, an alternative HIV-1 receptor. These findings elucidate potential antiviral functions of vaccine-elicited HIV-1 envelope-specific IgA that may act to block HIV-1 acquisition at the portal of entry by preventing HIV-1 binding to galactosylceramide and mediating antibody Fc receptor-mediated virion phagocytosis. Furthermore, these findings highlight the complex and diverse interactions of vaccine-elicited IgA with pathogens that depend on IgA fine specificity and form (e.g., multimeric or monomeric) in the systemic circulation and mucosal compartments.

IMPORTANCE
Host-pathogen interactions
in vivo
involve numerous immune mechanisms that can lead to pathogen clearance. Understanding the nature of antiviral immune mechanisms can inform the design of efficacious HIV-1 vaccine strategies. Evidence suggests that both neutralizing and nonneutralizing antibodies can mediate some protection against HIV in animal models. Although numerous studies have characterized the functional properties of HIV-1-specific IgG, more studies are needed on the functional attributes of HIV-1-specific IgA, specifically for vaccine-elicited IgA. Characterization of the functional properties of HIV-1 Env-specific IgA monoclonal antibodies from human vaccine clinical trials are critical toward understanding the capacity of the host immune response to block HIV-1 acquisition.
Therapeutic Efficacy of Vectored PGT121 Gene Delivery in HIV-1-Infected Humanized Mice
Authors: Badamchi-Zadeh Alexander , Tartaglia Lawrence J. , Abbink Peter , Bricault Christine A. , Liu Po-Ting , Boyd Michael , Kirilova Marinela , Mercado Noe B. , Nanayakkara Ovini S. , Vrbanac Vladimir D. , Tager Andrew M. , Larocca Rafael A. , Seaman Michael S. , Barouch Dan H.
Published: January 2018

Broadly neutralizing antibodies (bNAbs) are being explored for HIV-1 prevention and cure strategies. However, administration of purified bNAbs poses challenges in resource-poor settings, where the HIV-1 disease burden is greatest.
In vivo
vector-based production of bNAbs represents an alternative strategy. We investigated adenovirus serotype 5 (Ad5) and adeno-associated virus serotype 1 (AAV1) vectors to deliver the HIV-1-specific bNAb PGT121 in wild-type and immunocompromised C57BL/6 mice as well as in HIV-1-infected bone marrow-liver-thymus (BLT) humanized mice. Ad5.PGT121 and AAV1.PGT121 produced functional antibody
in vivo
. Ad5.PGT121 produced PGT121 rapidly within 6 h, whereas AAV1.PGT121 produced detectable PGT121 in serum by 72 h. Serum PGT121 levels were rapidly reduced by the generation of anti-PGT121 antibodies in immunocompetent mice but were durably maintained in immunocompromised mice. In HIV-1-infected BLT humanized mice, Ad5.PGT121 resulted in a greater reduction of viral loads than did AAV1.PGT121. Ad5.PGT121 also led to more-sustained virologic control than purified PGT121 IgG. Ad5.PGT121 afforded more rapid, robust, and durable antiviral efficacy than AAV1.PGT121 and purified PGT121 IgG in HIV-1-infected humanized mice. Further evaluation of vector delivery of HIV-1 bNAbs is warranted, although approaches to prevent the generation of antiantibody responses may also be required.

IMPORTANCE
Broadly neutralizing antibodies (bNAbs) are being explored for HIV-1 prevention and cure strategies, but delivery of purified antibodies may prove challenging. We investigated adenovirus serotype 5 (Ad5) and adeno-associated virus serotype 1 (AAV1) vectors to deliver the HIV-1-specific bNAb PGT121. Ad5.PGT121 afforded more rapid, robust, and durable antiviral efficacy than AAV1.PGT121 and purified PGT121 IgG in HIV-1-infected humanized mice.
Sustained Specific and Cross-Reactive T Cell Responses to Zika and Dengue Virus NS3 in West Africa
Authors: Herrera Bobby Brooke , Tsai Wen-Yang , Chang Charlotte A. , Hamel Donald J. , Wang Wei-Kung , Lu Yichen , Mboup Souleymane , Kanki Phyllis J.
Published: January 2018

Recent studies on the role of T cells in Zika virus (ZIKV) infection have shown that T cell responses to Asian ZIKV infection are important for protection, and that previous dengue virus (DENV) exposure amplifies the protective T cell response to Asian ZIKV. Human T cell responses to African ZIKV infection, however, remain unexplored. Here, we utilized the modified anthrax toxin delivery system to develop a flavivirus enzyme-linked immunosorbent spot (ELISPOT) assay. Using human ZIKV and DENV samples from Senegal, West Africa, our results demonstrate specific and cross-reactive T cell responses to nonstructural protein 3 (NS3). Specifically, we found that T cell responses to NS3 protease are ZIKV and DENV specific, but responses to NS3 helicase are cross-reactive. Sequential sample analyses revealed immune responses sustained many years after infection. These results have important implications for African ZIKV/DENV vaccine development, as well as for potential flavivirus diagnostics based on T cell responses.

IMPORTANCE
The recent Zika virus (ZIKV) epidemic in Latin America and the associated congenital microcephaly and Guillain-Barré syndrome have raised questions as to why we have not recognized these distinct clinical diseases in Africa. The human immunologic response to ZIKV and related flaviviruses in Africa represents a research gap that may shed light on the mechanisms contributing to protection. The goal of our study was to develop an inexpensive assay to detect and characterize the T cell response to African ZIKV and DENV. Our data show long-term specific and cross-reactive human immune responses against African ZIKV and DENV, suggesting the usefulness of a diagnostic based on the T cell response. Additionally, we show that prior flavivirus exposure influences the magnitude of the T cell response. The identification of immune responses to African ZIKV and DENV is of relevance to vaccine development.
Specific Mutations in the PB2 Protein of Influenza A Virus Compensate for the Lack of Efficient Interferon Antagonism of the NS1 Protein of Bat Influenza A-Like Viruses
Authors: Aydillo Teresa , Ayllon Juan , Pavlisin Amzie , Martinez-Romero Carles , Tripathi Shashank , Mena Ignacio , Moreira-Soto Andrés , Vicente-Santos Amanda , Corrales-Aguilar Eugenia , Schwemmle Martin , García-Sastre Adolfo
Published: January 2018

Recently, two new influenza A-like viruses have been discovered in bats, A/little yellow-shouldered bat/Guatemala/060/2010 (HL17NL10) and A/flat-faced bat/Peru/033/2010 (HL18NL11). The hemagglutinin (HA)-like (HL) and neuraminidase (NA)-like (NL) proteins of these viruses lack hemagglutination and neuraminidase activities, despite their sequence and structural homologies with the HA and NA proteins of conventional influenza A viruses. We have now investigated whether the NS1 proteins of the HL17NL10 and HL18NL11 viruses can functionally replace the NS1 protein of a conventional influenza A virus. For this purpose, we generated recombinant influenza A/Puerto Rico/8/1934 (PR8) H1N1 viruses containing the NS1 protein of the PR8 wild-type, HL17NL10, and HL18NL11 viruses. These viruses (r/NS1PR8, r/NS1HL17, and r/NS1HL18, respectively) were tested for replication in bat and nonbat mammalian cells and in mice. Our results demonstrate that the r/NS1HL17 and r/NS1HL18 viruses are attenuated
in vitro
and
in vivo
. However, the bat NS1 recombinant viruses showed a phenotype similar to that of the r/NS1PR8 virus in STAT1
−/−
human A549 cells and mice, both
in vitro
and
in vivo
systems being unable to respond to interferon (IFN). Interestingly, multiple mouse passages of the r/NS1HL17 and r/NS1HL18 viruses resulted in selection of mutant viruses containing single amino acid mutations in the viral PB2 protein. In contrast to the parental viruses, virulence and IFN antagonism were restored in the selected PB2 mutants. Our results indicate that the NS1 protein of bat influenza A-like viruses is less efficient than the NS1 protein of its conventional influenza A virus NS1 counterpart in antagonizing the IFN response and that this deficiency can be overcome by the influenza virus PB2 protein.

IMPORTANCE
Significant gaps in our understanding of the basic features of the recently discovered bat influenza A-like viruses HL17NL10 and HL18NL11 remain. The basic biology of these unique viruses displays both similarities to and differences from the basic biology of conventional influenza A viruses. Here, we show that recombinant influenza A viruses containing the NS1 protein from HL17NL10 and HL18NL11 are attenuated. This attenuation was mediated by their inability to antagonize the type I IFN response. However, this deficiency could be compensated for by single amino acid replacements in the PB2 gene. Our results unravel a functional divergence between the NS1 proteins of bat influenza A-like and conventional influenza A viruses and demonstrate an interplay between the viral PB2 and NS1 proteins to antagonize IFN.
HIV-1 Vif's Capacity To Manipulate the Cell Cycle Is Species Specific
Authors: Evans Edward L. , Becker Jordan T. , Fricke Stephanie L. , Patel Kishan , Sherer Nathan M.
Published: January 2018

Cells derived from mice and other rodents exhibit profound blocks to HIV-1 virion production, reflecting species-specific incompatibilities between viral Tat and Rev proteins and essential host factors cyclin T1 (CCNT1) and exportin-1 (XPO1, also known as CRM1), respectively. To determine if mouse cell blocks other than CCNT1 and XPO1 affect HIV's postintegration stages, we studied HIV-1
NL4-3
gene expression in mouse NIH 3T3 cells modified to constitutively express HIV-1-compatible versions of CCNT1 and XPO1 (3T3.CX cells). 3T3.CX cells supported both Rev-independent and Rev-dependent viral gene expression and produced relatively robust levels of virus particles, confirming that CCNT1 and XPO1 represent the predominant blocks to these stages. Unexpectedly, however, 3T3.CX cells were remarkably resistant to virus-induced cytopathic effects observed in human cell lines, which we mapped to the viral protein Vif and its apparent species-specific capacity to induce G
2
/M cell cycle arrest. Vif was able to mediate rapid degradation of human APOBEC3G and the PPP2R5D regulatory B56 subunit of the PP2A phosphatase holoenzyme in mouse cells, thus demonstrating that Vif
NL4-3
's modulation of the cell cycle can be functionally uncoupled from some of its other defined roles in CUL5-dependent protein degradation. Vif was also unable to induce G
2
/M cell cycle arrest in other nonhuman cell types, including cells derived from nonhuman primates, leading us to propose that one or more human-specific cofactors underpin Vif's ability to modulate the cell cycle.

IMPORTANCE
Cells derived from mice and other rodents exhibit profound blocks to HIV-1 replication, thus hindering the development of a low-cost small-animal model for studying HIV/AIDS. Here, we engineered otherwise-nonpermissive mouse cells to express HIV-1-compatible versions of two species-specific host dependency factors, cyclin T1 (CCNT1) and exportin-1 (XPO1) (3T3.CX cells). We show that 3T3.CX cells rescue HIV-1 particle production but, unexpectedly, are completely resistant to virus-induced cytopathic effects. We mapped these effects to the viral accessory protein Vif, which induces a prolonged G
2
/M cell cycle arrest followed by apoptosis in human cells. Combined, our results indicate that one or more additional human-specific cofactors govern HIV-1's capacity to modulate the cell cycle, with potential relevance to viral pathogenesis in people and existing animal models.
Kaposi's Sarcoma-Associated Herpesvirus K8 Is an RNA Binding Protein That Regulates Viral DNA Replication in Coordination with a Noncoding RNA
Authors: Liu Dongcheng , Wang Yan , Yuan Yan
Published: January 2018

Kaposi's sarcoma-associated herpesvirus (KSHV) lytic replication and constant primary infection of fresh cells are crucial for viral tumorigenicity. The virus-encoded bZIP family protein K8 plays an important role in viral DNA replication in both viral reactivation and
de novo
infection. The mechanism underlying the functional role of K8 in the viral life cycle is elusive. Here, we report that K8 is an RNA binding protein that also associates with many other proteins, including other RNA binding proteins. Many protein-protein interactions involving K8 are mediated by RNA. Using a UV
c
ross
-l
inking and
i
mmuno
p
recipitation (CLIP) procedure combined with high-throughput sequencing, RNAs that are associated with K8 in BCBL-1 cells were identified, including both viral (PAN, T1.4, T0.7, etc.) and cellular (MALAT-1, MRP, 7SK, etc.) RNAs. An RNA binding motif in K8 was defined, and mutation of the motif abolished the ability of K8 to bind to many noncoding RNAs, as well as viral DNA replication during
de novo
infection, suggesting that the K8 functions in viral replication are carried out through RNA association. The functions of K8 and associated T1.4 RNA were investigated in detail, and the results showed that T1.4 mediates the binding of K8 to ori-Lyt DNA. The T1.4-K8 complex physically bound to KSHV ori-Lyt DNA and recruited other proteins and cofactors to assemble a replication complex. Depletion of T1.4 abolished DNA replication in primary infection. These findings provide mechanistic insights into the role of K8 in coordination with T1.4 RNA in regulating KSHV DNA replication during
de novo
infection.

IMPORTANCE
Genomewide analyses of the mammalian transcriptome revealed that a large proportion of sequence previously annotated as noncoding regions is actually transcribed and gives rise to stable RNAs. The emergence of a large number of noncoding RNAs suggests that functional RNA-protein complexes, e.g., ribosomes or spliceosomes, are not ancient relics of the last ribo-organism but would be well adapted to a regulatory role in biology. K8 has been puzzling because of its unique characteristics, such as multiple regulatory roles in gene expression and DNA replication without DNA binding capability. This study reveals the mechanism underlying its regulatory role by demonstrating that K8 is an RNA binding protein that binds to DNA and initiates DNA replication in coordination with a noncoding RNA. It is suggested that many K8 functions, if not all, are carried out through its associated RNAs.
Two-year outcomes of intravitreal ziv-aflibercept
Authors: Mansour Ahmad M , Ashraf Mohammed , Charbaji Abdulrazzak , Younis Muhammad H , Souka Ahmed A , Dogra Avantika , Mansour Hana A , Chhablani Jay
Published: January 2018

Aim

To assess the two-year outcome of intravitreal ziv-aflibercept (IVZ) in eyes with macular diseases.

Methods

Consecutive subjects with various macular diseases that received six or more of 0.05 mL IVZ (1.25 mg) injections with at least 1 year follow-up were included. Outcome measures were best-corrected visual acuity (BCVA) (logarithm of the minimum angle of resolution) and central macular thickness (CMT) on spectral domain optical coherence tomography. Paired comparison was done using Wilcoxon signed-rank test calculator.

Results

107 eyes of 91 subjects received IVZ and were followed with mean±SD follow-up interval of 1.48±0.44 months following treat and extend or pro-re-nata protocol. The distribution included neovascular macular degeneration (42 eyes), diabetic macular oedema (32 eyes) and macular oedema secondary to retinal vein occlusion (11 eyes). Fifty eyes were naive, while 57 eyes were previously treated. Combining all disease categories, CMT decreased significantly by 133.0±153.0 µm at the 24-month follow-up (P<0.001) with BCVA gain of 0.35±0.37 at the 24-month follow-up (P<0.001) with mean number of injections of 8.5 at month 12, 2.4 between 12 and 18 month and 1.7 between 18 and 24 month. Ocular and systemic adverse effects included one episode of transient uveitis and one instance of central retinal artery occlusion after 1121 injections.

Conclusions

IVZ appears safe and efficacious in the therapy of macular diseases through 2 years.
Current strategies and successes in engaging women in vector control: a systematic review
Authors: Gunn Jayleen K L , Ernst Kacey C , Center Katherine E , Bischoff Kristi , Nuñez Annabelle V , Huynh Megan , Okello Amanda , Hayden Mary H
Published: January 2018

Introduction

Vector-borne diseases (VBDs) cause significant mortality and morbidity in low-income and middle-income countries and present a risk to high-income countries. Vector control programmes may confront social and cultural norms that impede their execution. Anecdotal evidence suggests that incorporating women in the design, delivery and adoption of health interventions increases acceptance and compliance. A better understanding of programmes that have attempted to increase women’s involvement in vector control could help shape best practices. The objective of this systematic review was to assess and critically summarise evidence regarding the effectiveness of women participating in vector control.

Methods

Seven databases were searched from inception to 21 December 2015. Two investigators independently reviewed all titles and abstracts for relevant articles. Grey literature was searched by assessing websites that focus on international development and vector control.

Results

In total, 23 articles representing 17 unique studies were included in this review. Studies discussed the involvement of women in the control of vectors for malaria (n=10), dengue (n=8), human African trypanosomiasis (n=3), schistosomiasis (n=1) and a combination (malaria and schistosomiasis, n=1). Seven programmes were found in the grey literature or through personal communications. Available literature indicates that women can be successfully engaged in vector control programmes and, when given the opportunity, they can create and sustain businesses that aim to decrease the burden of VBDs in their communities.

Conclusion

This systematic review demonstrated that women can be successfully engaged in vector control programmes at the community level. However, rigorous comparative effectiveness studies need to be conducted.
Making sense of child, early and forced marriage among Syrian refugee girls: a mixed methods study in Lebanon
Authors: Bartels Susan Andrea , Michael Saja , Roupetz Sophie , Garbern Stephanie , Kilzar Lama , Bergquist Harveen , Bakhache Nour , Davison Colleen , Bunting Annie
Published: January 2018

Introduction

The Syrian conflict has resulted in over 2.3 million child refugees in the Middle East and the prevalence of early marriage has reportedly increased among displaced Syrian families. This study explores the underlying factors contributing to child marriage among Syrian refugees in Lebanon with the goal of informing community-based strategies to address the issue.

Methods

In July–August 2016, trained interviewers collected self-interpreted stories in Lebanon using Cognitive Edge’s SenseMaker, a mixed-method data collection tool. Participants included married and unmarried Syrian girls, Syrian parents as well as married and unmarried men. Each participant shared a story about the experiences of Syrian girls and then interpreted the story by plotting their perspectives on a variety of questions. Patterns in the responses were analysed in SPSS and the accompanying qualitative narratives were reviewed to facilitate interpretation of the quantitative results.

Results

1422 self-interpreted stories from 1346 unique participants were collected with 40% of shared stories focused on (n=332) or mentioning (n=245) child marriage. Quantitative data summarised the different perspectives of female and male participants. Syrian girls and mothers were more likely to share stories about protection/security and/or education and were more likely to report that girls were overprotected. Male participants were more likely to share stories about financial security as well as sexual exploitation of girls and more often reported that girls were not protected enough. Despite these gendered perspectives, many of the shared narratives highlighted similar themes of financial hardship, lack of educational opportunities and safety concerns around sexual and gender-based violence (SGBV).

Conclusions

A complex myriad of factors contribute to early marriage including poverty, lack of educational opportunities and concerns about SGBV. Sexual exploitation under the guise of marriage is a reality for some Syrian girls. Gender-specific strategies to address child marriage might be more effective in reducing this harmful practice.
Examining changes in the equity of physician distribution in Japan: a specialty-specific longitudinal study
Authors: Hara Koji , Kunisawa Susumu , Sasaki Noriko , Imanaka Yuichi
Published: January 2018

Objectives

In this longitudinal study, we examined changes in the geographical distribution of physicians in Japan from 2000 to 2014 by clinical specialty with adjustments for healthcare demand based on population structure.

Methods

The Japanese population was adjusted for healthcare demand using health expenditure per capita stratified by age and sex. The numbers of physicians per 100 000 demand-adjusted population (DAP) in 2000 and 2014 were calculated for subprefectural regions known as secondary medical areas. Disparities in the geographical distribution of physicians for each specialty were assessed using Gini coefficients. A subgroup analysis was conducted by dividing the regions into four groups according to urban–rural classification and initial physician supply.

Results

Over the study period, the number of physicians per 100 000 DAP decreased in all specialties assessed (internal medicine: −6.9%, surgery: −26.0%, orthopaedics: −2.1%, obstetrics/gynaecology (per female population): −17.5%) except paediatrics (+33.3%) and anaesthesiology (+21.1%). No reductions in geographical disparity were observed in any of the specialties assessed. Geographical disparity increased substantially in internal medicine, surgery and obstetrics and gynaecology(OB/GYN). Rural areas with lower initial physician supply experienced the highest decreases in physicians per 100 000 DAP for all specialties assessed except paediatrics and anaesthesiology. In contrast, urban areas with lower initial physician supply experienced the lowest decreases in physicians per 100 000 DAP in internal medicine, surgery, orthopaedics and OB/GYN, but the highest increase in anaesthesiology.

Conclusion

Between 2000 and 2014, the number of physicians per 100 000 DAP in Japan decreased in all specialties assessed except paediatrics and anaesthesiology. There is also a growing urban–rural disparity in physician supply in all specialties assessed except paediatrics. Additional measures may be needed to resolve these issues and improve physician distribution in Japan.
Patients’ and therapists’ experiences with a new treatment programme for eating disorders that combines physical exercise and dietary therapy: the PED-t trial. A qualitative study protocol
Authors: Pettersen Gunn , Rosenvinge Jan H , Bakland Maria , Wynn Rolf , Mathisen Therese Fostervold , Sundgot-Borgen Jorunn
Published: January 2018

Introduction

Women with bulimia nervosa and binge eating disorder often suffer for many years before they seek professional help. Evidence-based treatments like cognitive–behavioural therapy (CBT) might be poorly accessible, and about 50% of those who receive CBT respond to it. Such outcome may reflect the heterogeneous nature of eating disorders, and addressing this heterogeneity calls for expanding the portfolio of treatment options. In particular, it is important to explore such options’ acceptability, tolerability and affordability expressed through experiences with the treatment. This protocol outlines the rationale and design of a qualitative study. It captures experiences from patients and therapists who were involved in a randomised controlled trial (RCT) exploring the efficacy of a new group-based treatment programme combining physical exercise and dietary therapy.

Methods and analysis

15 patients with bulimia nervosa or binge eating disorder, 10 therapists (physical trainers and dietitians) and 6–10 patients who dropped out of the RCT will be semistructurally interviewed. All interviews will be analysed using a systematic text condensation approach.

Ethics and dissemination

Results will be presented in peer-reviewed international journals, and at relevant international conferences. Key findings will be available to study participants as well as to patient organisations and health authorities. The overall study meets the intent and requirements of the Health Research Act and the Declaration of Helsinki. It is approved by the regional committee for medical research ethics (2013/1871).

Trial registration number

NCT02079935; Pre-results.
Cohort profile: effect of malaria in early pregnancy on fetal growth in Benin (RECIPAL preconceptional cohort)
Authors: Accrombessi Manfred , Yovo Emmanuel , Cottrell Gilles , Agbota Gino , Gartner Agnès , Martin-Prevel Yves , Fanou-Fogny Nadia , Djossinou Diane , Zeitlin Jennifer , Tuikue-Ndam Nicaise , Bodeau-Livinec Florence , Houzé Sandrine , Jackson Nicola , Ayemonna Paul , Massougbodji Achille , Cot Michel , Fievet Nadine , Briand Valérie
Published: January 2018

Purpose

REtard de Croissance Intra-uterin et PALudisme (RECIPAL) is an original preconceptional cohort designed to assess the consequences of malaria during the first trimester of pregnancy, which is a poorly investigated period in Africa and during which malaria may be detrimental to the fetus.

Participants

For this purpose, a total of 1214 women of reproductive age living in Sô-Ava and Akassato districts (south Benin) were followed up monthly from June 2014 to December 2016 until 411 of them became pregnant. A large range of health determinants was collected both before and during pregnancy from the first weeks of gestation to delivery. Five Doppler ultrasound scans were performed for early dating of the pregnancy and longitudinal fetal growth assessment.

Findings to date

Pregnant women were identified at a mean of 6.9 weeks of gestation (wg). Preliminary results confirmed the high prevalence of malaria in the first trimester of pregnancy, with more than 25.4% of women presenting at least one microscopic malarial infection during this period. Most infections occurred before six wg. The prevalence of low birth weight, small birth weight for gestational age (according to INTERGROWTH-21st charts) and preterm birth was 9.3%, 18.3% and 12.6%, respectively.

Future plans

REtard de Croissance Intra-uterin et PALudisme (RECIPAL) represents at this time a unique resource that will provide information on multiple infectious (including malaria), biological, nutritional and environmental determinants in relation to health outcomes in women of reproductive age, pregnant women and their newborns. It will contribute to better define future recommendations for the prevention of malaria in early pregnancy and maternal malnutrition in Africa. It confirms that it is possible to constitute a preconceptional pregnancy cohort in Africa and provides valuable information for researchers starting cohorts in the future.
Evaluating team decision-making as an emergent phenomenon
Authors: Kinnear John , Wilson Nick , O’Dwyer Anthony
Published: January 2018

Background

The complexity of modern clinical practice has highlighted the fallibility of individual clinicians’ decision-making, with effective teamwork emerging as a key to patient safety. Dual process theory is widely accepted as a framework for individual decision-making, with type 1 processes responsible for fast, intuitive and automatic decisions and type 2 processes for slow, analytical decisions. However, dual process theory does not explain cognition at the group level, when individuals act in teams. Team cognition resulting from dynamic interaction of individuals is said to be more resilient to decision-making error and greater than simply aggregated cognition.

Methods

Clinicians were paired as teams and asked to solve a cognitive puzzle constructed as a drug calculation. The frequency at which the teams made incorrect decisions was compared with that of individual clinicians answering the same question.

Results

When clinicians acted in pairs, 63% answered the cognitive puzzle correctly, compared with 33% of clinicians as individuals, showing a statistically significant difference in performance (χ2 (1, n=116)=24.329, P<0.001). Based on the predicted performance of teams made up of the random pairing of individuals who had the same propensity to answer as previously, there was no statistical difference in the actual and predicted teams’ performance.

Conclusions

Teams are less prone to making errors of decision-making than individuals. However, the improved performance is likely to be owing to the effect of aggregated cognition rather than any improved decision-making as a result of the interaction. There is no evidence of team cognition as an emergent and distinct entity.
Interstitial lung abnormalities and self-reported health and functional status
Authors: Axelsson Gisli Thor , Putman Rachel K , Araki Tetsuro , Sigurdsson Sigurdur , Gudmundsson Elias Freyr , Eiriksdottir Gudny , Aspelund Thor , Miller Ezra R , Launer Lenore J , Harris Tamara B , Hatabu Hiroto , Gudnason Vilmundur , Hunninghake Gary Matt , Gudmundsson Gunnar
Published: January 2018

We investigated the association between interstitial lung abnormalities (ILA) and self-reported measures of health and functional status in 5764 participants from the Age, Gene/Environment Susceptibility-Reykjavik study. The associations of ILA to activities of daily living (ADLs), general health status and physical activity were explored using logistic regression models. Participants with ILA were less likely to be independent in ADLs (OR 0.70; 95% CI 0.55 to 0.90) to have good or better self-reported health (OR 0.66; 95% CI 0.52 to 0.82) and to participate in physical activity (OR 0.72; CI 0.56 to 0.91). The results demonstrate ILA’s association with worsening self-reported health and functional status.
Change in viral bronchiolitis management in primary care in the UK after the publication of NICE guideline
Authors: Carande Elliott J , Cheung C Ronny , Pollard Andrew J , Drysdale Simon B
Published: January 2018

The aim of this study was to investigate changes in general practitioner (GP) management before and after the publication of the National Institute for Health and Care Excellence bronchiolitis guideline. In March 2015 and May 2016, an electronic questionnaire was sent to GPs. It was completed by 1001 GPs in 2015 and 1009 in 2016. There were small but significant improvements in proportions of GPs using a guideline, measuring oxygen saturations and providing written guidance, and appropriate reductions in those prescribing medications. Thirty-five per cent had read the guideline and 25% changed their practice since guideline publication. There were modest but significant improvements in reported management by GPs after guideline publication.
Novel dry cryotherapy system for cooling the equine digit
Authors: Morgan Jessica , Stefanovski Darko , Lenfest Margret , Chatterjee Sraboni , Orsini James
Published: January 2018

Objectives

Digital cryotherapy is commonly used for laminitis prophylaxis and treatment. Currently validated methods for distal limb cryotherapy involve wet application or compression technology. There is a need for a practical, affordable, dry cryotherapy method that effectively cools the digit. The objective of this study was to evaluate the hoof wall surface temperatures (HWSTs) achieved with a novel dry cryotherapy technology.

Design

Repeated‐measures in vivo experimental study.

Setting

Experimental intervention at a single site.

Participants

6 systemically healthy horses (3 mares, 3 geldings).

Interventions

Cryotherapy was applied to six horses for eight hours with a commercially available rubber and rubber and welded fabricice boot, which extended proximally to include the foot and pastern. Reusable malleable cold therapy packs were secured against the foot and pastern with the three built‐in hook‐and‐loop fastener panels.

Primary and secondary outcome measures

HWST and pastern surface temperature of the cryotherapy‐treated limb, HWST of the control limb and ambient temperature were recorded every five minutes throughout the study period.

Results

Results were analysed with mixed‐effects multivariable regression analysis. The HWST (median 11.1°C, interquartile range 8.6°C–14.7°C) in the cryotherapy‐treated limb was significantly decreased compared with the control limb (median 29.7°C, interquartile range 28.9°C–30.4°C) (P≤0.001). Cryotherapy limb HWST reached a minimum of 6.75°C (median) with an interquartile range of 4.1°C–9.3°C. Minimum HWST was achieved 68 minutes after cryotherapy pack application.

Conclusions

Dry application of cryotherapy significantly reduced HWST and reached minimums below the therapeutic target of 10°C. This cryotherapy method might offer an effective alternative for digital cooling.
Transportation of ostriches ‐ a review
Authors: Wotton S. B. , Hewitt L.
Published: January 2018

The transport of farm animals has been the subject of much research in recent years; however, the information available on ostrich transportation is limited. This paper draws together past and recent scientific literature and practical experience. The current state of knowledge is summarised and areas which require further research are identified.
Magnesium–zinc–calcium–vitamin D co-supplementation improves glycemic control and markers of cardiometabolic risk in gestational diabetes: a randomized, double-blind, placebo-controlled trial
Authors: Karamali Maryam , Bahramimoghadam Shahla , Sharifzadeh Fateme , Asemi Zatollah
Published: January 2018

To the best our knowledge, data on the effects of magnesium–zinc–calcium–vitamin D co-supplementation on glycemic control and markers of cardiometabolic risk in gestational diabetes mellitus (GDM) are scarce. The purpose of this study was to establish the effects of magnesium–zinc–calcium–vitamin D co-supplementation on glycemic control and markers of cardiometabolic risk of GDM patients. Sixty patients with GDM, aged 18–40 years, were randomized into 2 groups to intake either magnesium–zinc–calcium–vitamin D co-supplements or placebo (n = 30 each group) for 6 weeks in a randomized, double-blind, placebo-controlled trial. Fasting blood samples were taken at baseline and week 6 to quantify related markers. After the 6-week intervention, compared with the placebo, magnesium–zinc–calcium–vitamin D co-supplementation resulted in significant reductions in fasting plasma glucose (−0.37 ± 0.09 vs. +0.01 ± 0.09 mmol/L, P = 0.003), serum insulin levels (−21.0 ± 4.8 vs. +7.2 ± 4.8 pmol/L, P < 0.001), homeostatic model of assessment for insulin resistance (−1.0 ± 1.1 vs. +0.3 ± 1.3, P < 0.001), and a significant increase in quantitative insulin sensitivity check index (+0.02 ± 0.03 vs. −0.002 ± 0.03, P = 0.003). In addition, magnesium–zinc–calcium–vitamin D co-supplementation significantly decreased serum triglycerides (−0.25 ± 0.10 vs. +0.34 ± 0.10 mmol/L, P = 0.001) and very-low-density–cholesterol concentrations (−0.11 ± 0.04 vs. +0.15 ± 0.04 mmol/L, P = 0.001) compared with the placebo. Overall, the results of this study demonstrated that magnesium–zinc–calcium–vitamin D co-supplementation for 6 weeks among patients with GDM had beneficial effects on glycemic control and few markers of cardiometabolic risk.