bluuebunny/crossref_metadata_2025 huggingface dataset:
Antibodies that conformationally activate ADAMTS13 allosterically enhance metalloprotease domain function
Authors: Schelpe An-Sofie , Petri Anastasis , Roose Elien , Pareyn Inge , Deckmyn Hans , De Meyer Simon F. , Crawley James T. B. , Vanhoorelbeke Karen
Published: March 2020

Plasma ADAMTS13 circulates in a folded conformation that is stabilized by an interaction between the central Spacer domain and the C-terminal CUB (complement components C1r and C1s, sea urchin protein Uegf, and bone morphogenetic protein-1) domains. Binding of ADAMTS13 to the VWF D4(-CK) domains or to certain activating murine monoclonal antibodies (mAbs) induces a structural change that extends ADAMTS13 into an open conformation that enhances its function. The objective was to characterize the mechanism by which conformational activation enhances ADAMTS13-mediated proteolysis of VWF. The activating effects of a novel anti-Spacer (3E4) and the anti-CUB1 (17G2) mAbs on the kinetics of proteolysis of VWF A2 domain fragments by ADAMTS13 were analyzed. mAb-induced conformational changes in ADAMTS13 were investigated by enzyme-linked immunosorbent assay. Both mAbs enhanced ADAMTS13 catalytic efficiency (kcat/Km) by ∼twofold (3E4: 2.0-fold; 17G2: 1.8-fold). Contrary to previous hypotheses, ADAMTS13 activation was not mediated through exposure of the Spacer or cysteine-rich domain exosites. Kinetic analyses revealed that mAb-induced conformational extension of ADAMTS13 enhances the proteolytic function of the metalloprotease domain (kcat), rather than augmenting substrate binding (Km). A conformational effect on the metalloprotease domain was further corroborated by the finding that incubation of ADAMTS13 with either mAb exposed a cryptic epitope in the metalloprotease domain that is normally concealed when ADAMTS13 is in a closed conformation. We show for the first time that the primary mechanism of mAb-induced conformational activation of ADAMTS13 is not a consequence of functional exosite exposure. Rather, our data are consistent with an allosteric activation mechanism on the metalloprotease domain that augments active site function.
Efficient identification of multiple pathways: RNA-Seq analysis of livers from 56Fe ion irradiated mice
Authors: Nia Anna M. , Chen Tianlong , Barnette Brooke L. , Khanipov Kamil , Ullrich Robert L. , Bhavnani Suresh K. , Emmett Mark R.
Published: March 2020

Background

mRNA interaction with other mRNAs and other signaling molecules determine different biological pathways and functions. Gene co-expression network analysis methods have been widely used to identify correlation patterns between genes in various biological contexts (e.g., cancer, mouse genetics, yeast genetics). A challenge remains to identify an optimal partition of the networks where the individual modules (clusters) are neither too small to make any general inferences, nor too large to be biologically interpretable. Clustering thresholds for identification of modules are not systematically determined and depend on user-settable parameters requiring optimization. The absence of systematic threshold determination may result in suboptimal module identification and a large number of unassigned features.

Results

In this study, we propose a new pipeline to perform gene co-expression network analysis. The proposed pipeline employs WGCNA, a software widely used to perform different aspects of gene co-expression network analysis, and Modularity Maximization algorithm, to analyze novel RNA-Seq data to understand the effects of low-dose56Fe ion irradiation on the formation of hepatocellular carcinoma in mice. The network results, along with experimental validation, show that using WGCNA combined with Modularity Maximization, provides a more biologically interpretable network in our dataset, than that obtainable using WGCNA alone. The proposed pipeline showed better performance than the existing clustering algorithm in WGCNA, and identified a module that was biologically validated by a mitochondrial complex I assay.

Conclusions

We present a pipeline that can reduce the problem of parameter selection that occurs with the existing algorithm in WGCNA, for applicable RNA-Seq datasets. This may assist in the future discovery of novel mRNA interactions, and elucidation of their potential downstream molecular effects.
Phylogenetic relationship between Australian Fusarium oxysporum isolates and resolving the species complex using the multispecies coalescent model
Authors: Achari Saidi R. , Kaur Jatinder , Dinh Quang , Mann Ross , Sawbridge Tim , Summerell Brett A. , Edwards Jacqueline
Published: March 2020

Background

TheFusarium oxysporumspecies complex (FOSC) is a ubiquitous group of fungal species readily isolated from agroecosystem and natural ecosystem soils which includes important plant and human pathogens. Genetic relatedness within the complex has been studied by sequencing either the genes or the barcoding gene regions within those genes. Phylogenetic analyses have demonstrated a great deal of diversity which is reflected in the differing number of clades identified: three, five and eight. Genetic limitation within the species in the complex has been studied through Genealogical Concordance Phylogenetic Species Recognition (GCPSR) analyses with varying number of phylogenetic ‘species’ identified ranging from two to 21. Such differing views have continued to confuse users of these taxonomies.

Results

The phylogenetic relationships between AustralianF. oxysporumisolates from both natural and agricultural ecosystems were determined using three datasets: whole genome, nuclear genes, and mitochondrial genome sequences. The phylogenies were concordant except for three isolates. There were three concordant clades from all the phylogenies suggesting similar evolutionary history for mitochondrial genome and nuclear genes for the isolates in these three clades. Applying a multispecies coalescent (MSC) model on the eight single copy nuclear protein coding genes from the nuclear gene dataset concluded that the three concordant clades correspond to three phylogenetic species within the FOSC. There was 100% posterior probability support for the formation of three species within the FOSC. This is the first report of using the MSC model to estimate species within theF. oxysporumspecies complex. The findings from this study were compared with previously published phylogenetics and species delimitation studies.

Conclusion

Phylogenetic analyses using three different gene datasets from AustralianF. oxysporumisolates have all supported the formation of three major clades which delineated into three species. Species 2 (Clade 3) may be calledF. oxysporumas it contains the neotype forF. oxysporum.
Overexpression of GmNFYA5 confers drought tolerance to transgenic Arabidopsis and soybean plants
Authors: Ma Xiao-Jun , Yu Tai-Fei , Li Xiao-Hui , Cao Xin-You , Ma Jian , Chen Jun , Zhou Yong-Bin , Chen Ming , Ma You-Zhi , Zhang Jun-Hua , Xu Zhao-Shi
Published: March 2020

Background

Crop productivity is challenged by abiotic stresses, among which drought stress is the most common.NF-Ygenes, especiallyNF-YAgenes, regulate tolerance to abiotic stress.

Results

Soybean NF-Y geneGmNFYA5was identified to have the highest transcript level among all 21NF-YAgenes in soybean (Glycine maxL.) under drought stress. Drought-induced transcript ofGmNFYA5was suppressed by the ABA synthesis inhibitor naproxen (NAP).GmNFYA5transcript was detected in various tissues at vegetative and reproductive growth stages with higher levels in roots and leaves than in other tissues, which was consist with theGmNFYA5promoter: GUS fusion assay. Overexpression ofGmNFYA5in transgenic Arabidopsis plants caused enhanced drought tolerance in seedlings by decreasing stomatal aperture and water loss from leaves. Overexpression and suppression ofGmNFYA5in soybean resulted in increased and decreased drought tolerance, respectively, relative to plants with an empty vector (EV). Transcript levels of ABA-dependent genes (ABI2,ABI3,NCED3,LEA3,RD29A,P5CS1,GmWRKY46,GmNCED2andGmbZIP1) and ABA-independent genes (DREB1A,DREB2A,DREB2B,GmDREB1,GmDREB2andGmDREB3) in transgenic plants overexpressingGmNFYA5were higher than those of wild-type plants under drought stress; suppression ofGmNFYA5transcript produced opposite results. GmNFYA5 probably regulated the transcript abundance ofGmDREB2andGmbZIP1by binding to the promoters in vivo.

Conclusions

Our results suggested that overexpression ofGmNFYA5improved drought tolerance in soybean via both ABA-dependent and ABA-independent pathways.
Unusual accumulation of a wide array of antimicrobial resistance mechanisms in a patient with cytomegalovirus-associated hemophagocytic lymphohistiocytosis: a case report
Authors: Hasan Mohammad Rubayet , Sundaram Manu Somasundaram , Sundararaju Sathyavathi , Tsui Kin-Ming , Karim Mohammed Yousuf , Roscoe Diane , Imam Omar , Janahi Mohammad A. , Thomas Eva , Dobson Simon , Tan Rusung , Tang Patrick , Lopez Andres Perez
Published: March 2020

Background

Infections with multidrug-resistant organisms (MDRO) pose a serious threat to patients with dysregulated immunity such as in hemophagocytic lymphohistiocytosis (HLH), but such infections have rarely been comprehensively characterized. Here, we present a fatal case of HLH secondary to cytomegalovirus (CMV) infection complicated by both anti-viral drug resistance and sepsis from multiple MDROs including pandrug-resistant superbug bacteria.

Case presentation

A previously healthy, six-year-old boy presented with a 45-day history of fever prior to a diagnosis of hemophagocytic lymphohistiocytosis and hemorrhagic colitis, both associated with CMV. On hospital admission, the patient was found to be colonized with multiple, multidrug-resistant (MDR) bacteria including vancomycin-resistant enterococci (VRE) and carbapenamase-producing organisms (CPO). He eventually developed respiratory, urine and bloodstream infections with highly drug-resistant, including pandrug-resistant bacteria, which could not be controlled by antibiotic treatment. Antiviral therapy also failed to contain his CMV infection and the patient succumbed to overwhelming bacterial and viral infection. Whole genome sequencing (WGS) of the MDR bacteria and metagenomic analysis of his blood sample revealed an unusual accumulation of a wide range of antimicrobial resistance mechanisms in a single patient, including antiviral resistance to ganciclovir, and resistance mechanisms to all currently available antibiotics.

Conclusions

The case highlights both the risk of acquiring MDR superbugs and the severity of these infections in HLH patients.
Correction to: Clinical and genetic analysis of two wolfram syndrome families with high occurrence of wolfram syndrome and diabetes type II: a case report
Authors: Sobhani Maryam , Tabatabaiefar Mohammad Amin , Ghafouri-Fard Soudeh , Rajab Asadollah , Hojjat Asal , Kajbafzadeh Abdol-Mohammad , Noori-Daloii Mohammad Reza
Published: March 2020

Following publication of the original article [1], the authors flagged that the name of ‘Asal Hojjat’ was misspelled; the name had been spelled as ‘Asal Hojat’.
The relationship between sleep onset time and cardiometabolic biomarkers in Chinese communities: a cross-sectional study
Authors: Wang Liqun , Li Jiangping , Du Yong , Sun Ting , Na Li , Wang Zhizhong
Published: March 2020

Background

Late sleep onset time (SOT) is a common social phenomenon in modern society, and it was associated with a higher risk of obesity. However, the literature gap exists about the SOT and cardiometabolic biomarkers which closely associated with obesity. The present study aimed to explore the association of SOT with cardiometabolic biomarkers in Chinese communities.

Methods

A cross-sectional study enrolled a total of 2418 participants was conducted in Ningxia province of China. The cardiometabolic biomarkers included triglyceride, total cholesterol, high-density lipoprotein, low-density lipoprotein and fasting plasma glucose were measured quantitatively using the standard method. The SOT and sleep duration were acquired by a self-report questionnaire. The multiple mixed-effect linear regression model was employed to examine the association.

Results

Binary analysis found an inverse association of SOT with high-density lipoprotein (β = − 0.05, 95%CI: − 0.06, − 0.03), with 1 h delayed in SOT the high-density lipoprotein decreased 0.05 mmol/L. After controlling for demographic variables, health-related behaviors, and physical health covariates, late SOT was associated with a higher level of triglyceride (β = 0.12, 95%CI: 0.06, 0.18), a higher level of low-density lipoprotein (β = 0.06, 95% CI: 0.02, 0.09), and a lower level of high-density lipoprotein (β = − 0.05, 95% CI: − 0.06, − 0.03). when stratified by sleep duration (less than eight hours vs. eight and longer hours), a positive association between SOT and LDL (β = 0.08, 95% CI: 0.04, 0.12) was found among participants with sleep duration eight hours and longer.

Conclusion

Late sleep onset time with the negative effect on the cardiometabolic biomarkers, and individuals with late SOT coupled with longer sleep duration may take risk of a higher level of low-density lipoprotein which in turn lead to increase the risk of cardiovascular disease.
30+ years of media analysis of relevance to chronic disease: a scoping review
Authors: Rowbotham Samantha , Astell-Burt Thomas , Barakat Tala , Hawe Penelope
Published: March 2020

Background

Chronic, non-communicable diseases are a significant public health priority, requiring action at individual, community and population levels, and public and political will for such action. Exposure to media, including news, entertainment, and advertising media, is likely to influence both individual behaviours, and attitudes towards preventive actions at the population level. In recent years there has been a proliferation of research exploring how chronic diseases and their risk factors are portrayed across various forms of media. This scoping review aims to map the literature in this area to identify key themes, gaps, and opportunities for future research in this area.

Methods

We searched three databases (Medline, PsycINFO and Global Health) in July 2016 and identified 499 original research articles meeting inclusion criteria: original research article, published in English, focusing on media representations of chronic disease (including how issues are framed in media, impact or effect of media representations, and factors that influence media representations). We extracted key data from included articles and examined the health topics, media channels and methods of included studies, and synthesised key themes across studies.

Results

Our findings show that research on media portrayals of chronic disease increased substantially between 1985 and 2016. Smoking and nutrition were the most frequent health topics, and television and print were the most common forms of media examined, although, as expected, research on online and social media channels has increased in recent years. The majority of studies focused on the amount and type of media coverage, including how issues are framed, typically using content analysis approaches. In comparison, there was much less research on the influences on and consequences of media coverage related to chronic disease, suggesting an important direction for future work.

Conclusions

The results highlight key themes across media research of relevance to chronic disease. More in-depth syntheses of studies within the identified themes will allow us to draw out the key patterns and learnings across the literature.
Complete regression of pulmonary squamous carcinoma in IPF following gemcitabine plus cisplatin: a case report and literature review
Authors: Ma Weirong , Li Hui , Tian Zhigang , Wang Shaojin , Zheng Xiwei , Hou Jia
Published: March 2020

Background

Lung cancer is one of the most common co-morbid conditions in patients with idiopathic pulmonary fibrosis (IPF) and negatively affects the prognosis of IPF; Current guidelines for the management of IPF do not give a clear statement on how to manage these patients, and traditional chemotherapy for lung cancer had a limited efficiency rate. Here, we present a rare case of primary lung squamous carcinoma in a patient with IPF whose tumor completely regressed following gemcitabine plus cisplatin therapy; the cancer was no longer detectable after 2 years upon follow-up.

Case presentation

Sixty-seven year-old male patient with IPF was admitted to hospital due to acute onset hemoptysis. In addition to a definite usual interstitial pneumonia (UIP) pattern, a chest CT scan showed a non-enhancing nodular opacity in the right upper lobe and an enhancing nodule in the right lower lobe. Bronchoscopic biopsy of the nodule in the right lower lobe revealed squamous lung cancer. After 2 cycles of chemotherapy with gemcitabine and cisplatin, the tumor in the right lower lobe was no longer detectable after 2 years of follow-up; however, the nodule in the right upper lobe had increased significantly. Finally, Mycobacterium tuberculosis (MTB) was cultured from the bronchoalveolar (BAL) sample submitted at the last evaluation, and the patient was confirmed to have active pulmonary TB.

Conclusion

We report the first documented case of complete pulmonary squamous carcinoma regression in IPF following gemcitabine plus cisplatin. Traditional chemotherapy is considered inadequate to cause the resulting regression of the tumor. The concomitant active pulmonary tuberculosis possibly underlies the mechanism.
Trends in life expectancy: did the gap between the healthy and the ill widen or close?
Authors: Meyer Anna C. , Drefahl Sven , Ahlbom Anders , Lambe Mats , Modig Karin
Published: March 2020

Background

During the past decades, life expectancy has continued to increase in most high-income countries. Previous research suggests that improvements in life expectancy have primarily been driven by advances at the upper end of the health distribution, while parts of the population have lagged behind. Using data from the entire Swedish population, this study aims to examine the life expectancy development among subgroups of individuals with a history of common diseases relative to that of the general population.

Methods

The remaining life expectancy at age 65 was estimated for each year in 1998–2017 among individuals with a history of disease, and for the total Swedish population. We defined population subgroups as individuals with a history of myocardial infarction, ischemic or hemorrhagic stroke, hip fracture, or colon, breast, or lung cancer. We further distinguished between different educational levels and Charlson comorbidity index scores.

Results

Life expectancy gains have been larger for men and women with a history of myocardial infarction, ischemic or hemorrhagic stroke, and colon or breast cancer than for the general population. The life expectancy gap between individuals with a history of hip fracture or lung cancer and the general population has, however, been growing. Education and comorbidity have affected mortality levels, but have not altered the rate of increase in life expectancy among individuals with disease history. The female advantage in life expectancy was less pronounced among individuals with disease history than among the general population.

Conclusions

Life expectancy has increased faster in many subpopulations with a history of disease than in the general population, while still remaining at lower levels. Improvements in life expectancy have been observed regardless of comorbidity or educational level. These findings suggest that the rise in overall life expectancy reflects more than just improved survival among the healthy or the delayed onset of disease.
Topical treatment with SPHINGOLIPIDS and GLYCOSAMINOGLYCANS for canine atopic dermatitis
Authors: Marsella Rosanna , Segarra Sergi , Ahrens Kim , Alonso Cristina , Ferrer Lluís
Published: March 2020

Background

Skin barrier dysfunction plays a key role in atopic dermatitis (AD). This impairment is related to altered composition and metabolism of epidermal sphingolipids and a deficiency of ceramides. Glycosaminoglycans (GAGs), and especially hyaluronic acid, could be useful in the management of AD. This study aimed to evaluate the effects of a novel topical treatment consisting of sphingolipids and GAGs extracts in dogs with AD. This formulation is different from previously tested products because the sphingolipid extract contained high amounts of sphingomyelin, a precursor of ceramides, and this has been shown to enhance endogenous synthesis of ceramides and to increase lamellar-related structures in vitro. Thus, it was hypothesized that this formulation could improve clinical disease and skin barrier function in patients with AD.

Results

Twelve house dust mite (HDM) allergic atopic beagle dogs were randomized into two groups: control (n = 6; no treatment) or treatment (n = 6; topical sphingolipids and GAGs twice weekly for 8 weeks). Dogs were challenged with allergen twice weekly and the severity of dermatitis was scored using the canine atopic dermatitis and extent severity index (CADESI-03) once weekly. Skin barrier function (measurement of transepidermal water loss) and severity of pruritus (both pruritus visual analog scale [PVAS] and pruritus timed episodes) were assessed at 0, 4 and 8 weeks of treatment. Assessments were done by personnel unaware of group allocation. Complete blood count, serum biochemistry and stratum corneum (SC) lipidomics analyses were done at baseline and at week 8.

Compared to baseline, significant increases in CADESI (P = 0.0003) and PVAS (P = 0.041) were observed only in the control group, and SC polyunsaturated fatty acids increased significantly only with treatment (P = 0.039). Compared to control, treatment group had a significantly lower CADESI after 1 week (P = 0.0078) and a significantly lower PVAS after 8 weeks (P = 0.0448). Treatment was well tolerated.

Conclusions

In this study in dogs with AD, a new topical formulation containing sphingomyelin-rich sphingolipids plus GAGs extracts attenuated the clinical worsening induced by HDM, supporting its use in atopic patients, either as an adjunctive treatment or used as monotherapy in certain cases.
Management of otitis externa with an led-illuminated gel: a randomized controlled clinical trial in dogs
Authors: Tambella Adolfo Maria , Attili Anna Rita , Beribè Francesca , Galosi Margherita , Marchegiani Andrea , Cerquetella Matteo , Palumbo Piccionello Angela , Vullo Cecilia , Spaterna Andrea , Fruganti Alessandro
Published: March 2020

Background

Canine otitis externa is a painful condition which can be challenging to treat due to difficulties in the administration of otic medication. This can be due to lack of owner compliance in the application of ear drops or due to the resentment that some dogs demonstrate when attempts are made to administer topical medication into a sensitive ear canal. The aim of the study was to assess the efficacy of a topical LED-illuminated gel (LIG) in canine otitis externa in comparison to standard of care therapy. Dogs with spontaneous otitis externa were randomly allocated in three groups: groups QW received LIG once weekly; BW received LIG twice weekly; group C received enrofloxacin and silver sulfadiazine twice daily. LIG consists of a topical application of a gel containing chromophores that, when illuminated by a LED lamp, re-emit fluorescent light which can stimulate physiological responses, promoting healing and controlling bacteria. The evaluation protocol (T0 to T5) considered clinical assessment (OTIS-3-index-scoring-system; pruritus-severity-scale; pain-severity-score; aural temperature), cytological scoring system, quali-quantitative bacteriologic assessment.

Results

All groups (QW, n = 21; BW, n = 23; C, n = 20) showed improvement during the study (QW: P < 0.02 for cytological and pain scores, P < 0.003 for bacteriologic assessment, P < 10− 4 for pruritus, total OTIS-3 and temperature assessments; BW: P < 10− 4 for all clinical, cytological and bacteriologic assessments; C: P < 0.02 for all clinical and cytological assessments, P < 10− 4 for bacteriologic assessment). The highest clinical score reduction occurred in Group BW (P < 0.014 in T3; P < 0.001 in T4 and P < 10− 4 in T5). BW reached the clinically relevant effect level at T3 (− 3.26 ± 1.21 levels), QW reached it at T4 (− 3.24 ± 0.99), C did not reach it. No differences between groups were seen in the reduction of CFU/mL (T0-T5).

Conclusions

All treatment groups showed a positive clinical effect. LIG administered twice-a-week was the most favourable protocol of the study. LIG may be considered beneficial in the management of canine otitis externa; it seems to be effective in controlling the clinical condition, including the signs of inflammation and local pain, the bacterial growth, and it may help increasing treatment compliance.
Sputum and blood transcriptomics characterisation of the inhaled PDE4 inhibitor CHF6001 on top of triple therapy in patients with chronic bronchitis
Authors: Govoni Mirco , Bassi Michele , Vezzoli Stefano , Lucci Germano , Emirova Aida , Nandeuil Marie Anna , Petruzzelli Stefano , Jellema Gera L. , Afolabi Ebenezer K. , Colgan Brendan , Leaker Brian , Kornmann Oliver , Beeh Kai Michael , Watz Henrik , Singh Dave
Published: March 2020

Background

Although phosphodiesterase-4 (PDE4) inhibitors have been shown to reduce COPD exacerbation rate, their biological mechanism of action is not completely elucidated at the molecular level. We aimed to characterise the whole genome gene expression profile of the inhaled PDE4-inhibitor CHF6001 on top of triple therapy in sputum cells and whole blood of patients with COPD and chronic bronchitis.

Methods

Whole genome gene expression analysis was carried out by microarray in 54 patients before and after 32 days treatment with CHF6001 800 and 1600 μg and placebo twice daily (BID) in a randomised crossover study.

Results

CHF6001 had a strong effect in sputum, with 1471 and 2598 significantly differentially-expressed probe-sets relative to placebo (p-adjusted for False Discovery Rate < 0.05) with 800 and 1600 μg BID, respectively. Functional enrichment analysis showed significant modulation of key inflammatory pathways involved in cytokine activity, pathogen-associated-pattern-recognition activity, oxidative stress and vitamin D with associated inhibition of downstream inflammatory effectors. A large number of pro-inflammatory genes coding for cytokines and matrix-metalloproteinases were significantly differentially expressed for both doses; the majority (> 87%) were downregulated, including macrophage inflammatory protein-1-alpha and 1-beta, interleukin-27-beta, interleukin-12-beta, interleukin-32, tumour necrosis factor-alpha-induced-protein-8, ligand-superfamily-member-15, and matrix-metalloproteinases-7,12 and 14. The effect in blood was not significant.

Conclusions

Inhaled PDE4 inhibition by CHF6001 on top of triple therapy in patients with COPD and chronic bronchitis significantly modulated key inflammatory targets and pathways in the lung but not in blood. Mechanistically these findings support a targeted effect in the lung while minimising unwanted systemic class-effects.

Trial registration

ClinicalTrial.gov, EudraCT, 2015–005550-35. Registered 15 July 2016.
Synthesis of acridone derivatives via heterologous expression of a plant type III polyketide synthase in Escherichia coli
Authors: Choi Gyu-Sik , Choo Hye Jeong , Kim Bong-Gyu , Ahn Joong-Hoon
Published: March 2020

Background

Acridone alkaloids are heterocyclic compounds that exhibit a broad-range of pharmaceutical and chemotherapeutic activities, including anticancer, antiviral, anti-inflammatory, antimalarial, and antimicrobial effects. Certain plant species such as Citrus microcarpa, Ruta graveolens, and Toddaliopsis bremekampii synthesize acridone alkaloids from anthranilate and malonyl-CoA.

Results

We synthesized two acridones in Escherichia coli. Acridone synthase (ACS) and anthraniloyl-CoA ligase genes were transformed into E. coli, and the synthesis of acridone was examined. To increase the levels of endogenous anthranilate, we tested several constructs expressing proteins involved in the shikimate pathway and selected the best construct. To boost the supply of malonyl-CoA, genes coding for acetyl-coenzyme A carboxylase (ACC) from Photorhabdus luminescens were overexpressed in E. coli. For the synthesis of 1,3-dihydroxy-10-methylacridone, we utilized an N-methyltransferase gene (NMT) to supply N-methylanthranilate and a new N-methylanthraniloyl-CoA ligase. After selecting the best combination of genes, approximately 17.3 mg/L of 1,3-dihydroxy-9(10H)-acridone (DHA) and 26.0 mg/L of 1,3-dihydroxy-10-methylacridone (NMA) were synthesized.

Conclusions

Two bioactive acridone derivatives were synthesized by expressing type III plant polyketide synthases and other genes in E. coli, which increased the supplement of substrates. This study showed that is possible to synthesize diverse polyketides in E. coli using plant polyketide synthases.
Mobile health clinics in the United States
Authors: Malone Nelson C. , Williams Mollie M. , Smith Fawzi Mary C. , Bennet Jennifer , Hill Caterina , Katz Jeffrey N. , Oriol Nancy E.
Published: March 2020

Background

Mobile health clinics serve an important role in the health care system, providing care to some of the most vulnerable populations. Mobile Health Map is the only comprehensive database of mobile clinics in the United States. Members of this collaborative research network and learning community supply information about their location, services, target populations, and costs. They also have access to tools to measure, improve, and communicate their impact.

Methods

We analyzed data from 811 clinics that participated in Mobile Health Map between 2007 and 2017 to describe the demographics of the clients these clinics serve, the services they provide, and mobile clinics’ affiliated institutions and funding sources.

Results

Mobile clinics provide a median number of 3491 visits annually. More than half of their clients are women (55%) and racial/ethnic minorities (59%). Of the 146 clinics that reported insurance data, 41% of clients were uninsured while 44% had some form of public insurance. The most common service models were primary care (41%) and prevention (47%). With regards to organizational affiliations, they vary from independent (33%) to university affiliated (24%), while some (29%) are part of a hospital or health care system. Most mobile clinics receive some financial support from philanthropy (52%), while slightly less than half (45%) receive federal funds.

Conclusion

Mobile health care delivery is an innovative model of health services delivery that provides a wide variety of services to vulnerable populations. The clinics vary in service mix, patient demographics, and relationships with the fixed health system. Although access to care has increased in recent years through the Affordable Care Act, barriers continue to persist, particularly among populations living in resource-limited areas. Mobile clinics can improve access by serving as a vital link between the community and clinical facilities. Additional work is needed to advance availability of this important resource.
Can traditional bonesetters become trained technicians? Feasibility study among a cohort of Nigerian traditional bonesetters
Authors: Onyemaechi Ndubuisi Onu , Itanyi Ijeoma Uchenna , Ossai Paulinus Okechukwu , Ezeanolue Echezona Edozie
Published: March 2020

Background

Traditional bonesetters (TBS) provide the majority of primary fracture care in Nigeria and other low- and middle-income countries (LMICs). They are widely patronized and their services are commonly associated with complications. The aim of the study was to establish the feasibility of formal training of TBS and subsequent integration into the healthcare system.

Methods

Two focus group discussions were conducted involving five TBS and eight orthopaedic surgeons in Enugu Nigeria. Audio-recordings made during the focus groups were transcribed verbatim and analysed using a thematic analysis method.

Results

Four themes were identified: Training of TBS, their experiences and challenges; perception of traditional bonesetting by orthopaedic surgeons; need for formal training TBS and willingness to offer and accept formal training to improve TBS practice. Participants (TBS group) acquired their skills through informal training by apprenticeship from relatives and family members. They recognized the need to formalize their training and were willing to accept training support from orthopaedists. The orthopaedists recognized that the TBS play a vital role in filling the gap created by shortage of orthopaedic surgeons and are willing to provide training support to them.

Conclusion

This study demonstrates the feasibility of providing formal training to TBS by orthopaedic surgeons to improve the quality of services and outcomes of TBS treatment. This is critical for integration of TBS into the primary healthcare system as orthopaedic technicians. Undoubtedly, this will transform the trauma system in Nigeria and other LMICs where TBS are widely patronized.
FXR-mediated inhibition of autophagy contributes to FA-induced TG accumulation and accordingly reduces FA-induced lipotoxicity
Authors: Wu Kun , Zhao Tao , Hogstrand Christer , Xu Yi-Chuang , Ling Shi-Cheng , Chen Guang-Hui , Luo Zhi
Published: March 2020

Background

Excessive dietary fat intake induces lipid deposition and contributes to the progress of nonalcoholic fatty liver disease (NAFLD). However, the underlying mechanisms are still unclear.

Methods

Yellow catfish were given two experimental diets with dietary lipid levels of 11.3 and 15.4%, respectively, for 56 days, and the contents of triglyceride (TG), nonesterified free fatty acids (NEFA) and bile acid (BA), RNA-seq, enzymatic activities and mRNA expression were deteremined in the liver tissues. Hepatocytes from yellow catfish liver tissues were isolated and cultured. Fatty acids (FA) (palmitic acid: OA, oleic acid =1:1), pathway inhibitors (MA, autophagy inhibitor; guggulsterone, FXR inhibitor) and agonist (rapamyicn, autophagy agonist; GW4064, FXR agonist) were used to incubate the cells. TG and NEFA contents, ultrastructural observation, autophagic vesicles and intracellular LD,apoptosis,western blot and Co-IP, and Immunofluorescence analysis, enzymatic activities and Q-PCR were decided.

Results

Using RNA sequencing, we found that high fat diets induced changes in expression of many genes associated with the pathways of lipid metabolism and autophagy. The mRNA profiles of the differentially expressed genes (DEG) indicated that high dietary fat-induced lipid deposition was predominantly influenced by the inhibition of autophagy. Using primary hepatocytes, we found that fatty acids (FA) suppressed autophagy, which in turn reduced cellular free FA level by decreasing triglyceride (TG) breakdown. Moreover, our study indicated that farnesoid X receptor (FXR)-cyclic AMP-responsive element-binding protein (CREB) axis was the pivotal physiological switch regulating FA-induced changes of autophagy and lipid metabolism, which represented cellular defenses against FA-induced lipotoxicity.

Conclusion

This discovery may provide new targets for treating pathological changes involved in the dysfunction of autophagy and metabolism, including NAFLD.

Graphical abstract
Advanced oxidation protein products induce microglia-mediated neuroinflammation via MAPKs-NF-κB signaling pathway and pyroptosis after secondary spinal cord injury
Authors: Liu Zhongyuan , Yao Xinqiang , Jiang Wangsheng , Li Wei , Zhu Siyuan , Liao Congrui , Zou Lin , Ding Ruoting , Chen Jianting
Published: March 2020

Background

Inflammatory response mediated by oxidative stress is considered as an important pathogenesis of spinal cord injury (SCI). Advanced oxidation protein products (AOPPs) are novel markers of oxidative stress and their role in inflammatory response after SCI remained unclear. This study aimed to investigate the role of AOPPs in SCI pathogenesis and explore the possible underlying mechanisms.

Methods

A C5 hemi-contusion injury was induced in Sprague-Dawley rats to confirm the involvement of AOPPs after SCI. For in vivo study, apocynin, the NADPH oxidase inhibitor was used to study the neuroprotective effects after SCI. For in vitro study, the BV2 microglia cell lines were pretreated with or without the inhibitor or transfected with or without small interference RNA (siRNA) and then stimulated with AOPPs. A combination of molecular and histological methods was used to clarify the mechanism and explore the signaling pathway both in vivo and in vitro. One-way analysis of variance (ANOVA) was conducted with Bonferroni post hoc tests to examine the differences between groups.

Results

The levels of AOPPs in plasma and cerebrospinal fluid as well as the contents in the spinal cord showed significant increase after SCI. Meanwhile, apocynin ameliorated tissue damage in the spinal cord after SCI, improving the functional recovery. Immunofluorescence staining and western blot analysis showed activation of microglia after SCI, which was in turn inhibited by apocynin. Pretreated BV2 cells with AOPPs triggered excessive generation of reactive oxygen species (ROS) by activating NADPH oxidase. Increased ROS induced p38 MAPK and JNK phosphorylation, subsequently triggering nuclear translocation of NF-κB p65 to express pro-inflammatory cytokines. Also, treatment of BV2 cells with AOPPs induced NLRP3 inflammasome activation and cleavage of Gasdermin-d (GSDMD), causing pyroptosis. This was confirmed by cleavage of caspase-1, production of downstream mature interleukin (IL)-1β and IL-18 as well as rupture of rapid cell membrane.

Conclusions

Collectively, these data indicated AOPPs as biomarkers of oxidative stress, modulating inflammatory response in SCI by multiple signaling pathways, which also included the induction of NADPH oxidase dependent ROS, and NLRP3-mediated pyroptosis, and activation of MAPKs and NF-κB.
Characterization of astrocytes throughout life in wildtype and APP/PS1 mice after early-life stress exposure
Authors: Abbink Maralinde R. , Kotah Janssen M. , Hoeijmakers Lianne , Mak Aline , Yvon-Durocher Genevieve , van der Gaag Bram , Lucassen Paul J. , Korosi Aniko
Published: March 2020

Background

Early-life stress (ES) is an emerging risk factor for later life development of Alzheimer’s disease (AD). We have previously shown that ES modulates amyloid-beta pathology and the microglial response to it in the APPswe/PS1dE9 mouse model. Because astrocytes are key players in the pathogenesis of AD, we studied here if and how ES affects astrocytes in wildtype (WT) and APP/PS1 mice and how these relate to the previously reported amyloid pathology and microglial profile.

Methods

We induced ES by limiting nesting and bedding material from postnatal days (P) 2–9. We studied in WT mice (at P9, P30, and 6 months) and in APP/PS1 mice (at 4 and 10 months) (i) GFAP coverage, cell density, and complexity in hippocampus (HPC) and entorhinal cortex (EC); (ii) hippocampal gene expression of astrocyte markers; and (iii) the relationship between astrocyte, microglia, and amyloid markers.

Results

In WT mice, ES increased GFAP coverage in HPC subregions at P9 and decreased it at 10 months. APP/PS1 mice at 10 months exhibited both individual cell as well as clustered GFAP signals. APP/PS1 mice when compared to WT exhibited reduced total GFAP coverage in HPC, which is increased in the EC, while coverage of the clustered GFAP signal in the HPC was increased and accompanied by increased expression of several astrocytic genes. While measured astrocytic parameters in APP/PS1 mice appear not be further modulated by ES, analyzing these in the context of ES-induced alterations to amyloid pathology and microglial shows alterations at both 4 and 10 months of age.

Conclusions

Our data suggest that ES leads to alterations to the astrocytic response to amyloid-β pathology.
Treating AO/OTA 44B lateral malleolar fracture in patients over 50 years of age: periarticular locking plate versus non-locking plate
Authors: Shih Chien-An , Jou I-Ming , Lee Pei-Yuan , Lu Chin-Li , Su Wei-Ren , Yeh Ming-Long , Wu Po-Ting
Published: March 2020

Background

The role of locking plate in lateral malleolar fracture fixation for the elderly remains unclear. The aim of our study is to compare radiological and functional outcomes in older patients (> 50 years) with AO/OTA 44B lateral malleolar fractures after locking plate (PLP) or one-third non-locking tubular plate (TP) lateral fixation.

Methods

We retrospectively reviewed the medical records of 72 patients (PLP group, 34 patients; TP group, 38 patients; mean age, 61.9 ± 7.6 years; range, 51–80 years; follow-up, 1 year). Patients with open fractures, syndesmosis injuries, and a previous ankle trauma or surgery were excluded. Demographic data, union rate, complications, radiographic outcomes, visual analog scale (VAS) scores, and foot and ankle outcome scores (FAOSs) between the groups were recorded and compared. We also investigated the association of clinical features with pain and function. Statistically, the Fisher’s exact test was used for categorical variables and the Mann-Whitney U test for the continuous variables. The final model for the multiple regression analysis was used to predict factors related to functional outcomes.

Results

There were no significant between-group differences in demographic data, complication rates, immediately postoperative distal fibula lengths, ankle osteoarthritis (OA) grades, talar tilt angles (TTAs) ≥ 2°, or reduction accuracy. All fractures achieved union. The PLP group had significantly lower rates of distal screw loosening, fibula shortening > 2 mm, OA grade progression, and TTAs ≥ 2°, and better FAOSs and VAS scores than was the case for the TP group after 1 year of follow-up (all p < 0.05). The severity of OA, TTA ≥ 2°, and distal screw loosening were positively associated with VAS scores, and negatively associated with FAOSs.

Conclusions

When treating AO/OTA 44B fractures in patients over 50 years of age, PLPs provided better VAS scores, FAOSs, and radiological outcomes, including less fibula shortening > 2 mm, less osteoarthritic (OA) ankle progression, less implant removal rate, and fewer TTAs ≥ 2° than was the case for TPs after a 1-year follow-up.

Level of evidence

Therapeutic level III
A comparison between arthroscopic and open surgery for treatment outcomes of chronic lateral ankle instability accompanied by osteochondral lesions of the talus
Authors: Xu Can , Li Mingqing , Wang Chenggong , Liu Hua
Published: March 2020

Background

This study aimed to examine the efficacy and safety of the arthroscopic treatment of osteochondral lesion of talus (OLT) and lateral ankle instability. It was hypothesized that the outcome of all-arthroscopic surgery was no worse than that of the combined open and arthroscopic surgery for treating chronic lateral ankle instability accompanied by OLT.

Methods

The patients diagnosed of chronic lateral ankle instability accompanied by OLT, who were surgically treated between May 2015 and May2017, were targeted for inclusion. Specifically, patients who received the arthroscopic treatment of OLT and lateral ankle instability were enrolled in the all-arthroscopic group, while patients who received the arthroscopic treatment of OLT and open lateral ankle stabilization were enrolled in the combined open and arthroscopic group. All the patients were followed up in terms of the Karlsson Ankle Functional Score, visual analog scale (VAS) score, Tegner activity score, and American Orthopaedic Foot & Ankle Society (AOFAS) score. Meanwhile, the satisfaction and complication rates were evaluated and compared.

Results

This retrospective study included a total of 67 patients, including 32 patients in the all-arthroscopic group and 35 patients in the combined group. At a minimum of 24-month follow-up, the functional outcomes were significantly improved in both groups in relation to the preoperative condition. However, the two groups did not differ significantly from each other in terms of the Karlsson score (83.1 ± 8.2 vs 81.7 ± 9.1;P= 0.89), the VAS score (1.8 ± 1.6 vs 2.1 ± 1.7;P= 0.73), the Tegner score (5.5 ± 2.3 vs 5.0 ± 2.1;P= 0.72), and the AOFAS score (87.7 ± 7.6 vs 86.9 ± 7.3;P= 0.77). In addition, the satisfaction and complication rates exhibited no significant differences between the two groups.

Conclusion

In comparison with the open lateral ankle stabilization and arthroscopic treatment of OLT, the all-arthroscopic procedure showed no difference in clinical outcomes at a minimum of 24-month follow-up. Despite the benefits of minimally invasive arthroscopic procedure combined with a relatively aggressive postoperative rehabilitation protocol, the clinical outcomes for patients with chronic lateral ankle instability accompanied by OLT did not yield significant improvement.

Trial registration

The present study was carried out with the approval issued by the Institutional Review Board of Xiangya Hospital (no. 202002010).
Correction to: Comparison of the effectiveness of a tailored cognitive behavioural therapy with a supportive listening intervention for depression in those newly diagnosed with multiple sclerosis (the ACTION-MS trial): protocol of an assessor-blinded, active comparator, randomised controlled trial
Authors: Kiropoulos Litza , Kilpatrick Trevor , Kalincik Tomas , Churilov Leonid , McDonald Elizabeth , Wijeratne Tissa , Threader Jennifer , Rozenblat Vanja , O’Brien-Simpson Neil , Van Der Walt Anneke , Taylor Lisa
Published: March 2020

After publication of our article [1] the authors have notified us that three of the names have been incorrectly spelled.
When assessing generalisability, focusing on differences in population or setting alone is insufficient
Authors: Burchett Helen E. D. , Kneale Dylan , Blanchard Laurence , Thomas James
Published: March 2020

Generalisability is typically only briefly mentioned in discussion sections of evaluation articles, which are unhelpful in judging whether an intervention could be implemented elsewhere, with similar effects. Several tools to assess generalisability exist, but they are difficult to operationalise and are rarely used. We believe a different approach is needed. Instead of focusing on similarities (or more likely, differences) in generic population and setting characteristics, generalisability assessments should focus on understanding an intervention’s mechanism of action - why or how an intervention was effective. We believe changes are needed to four types of research. First, outcome evaluations should draw on programme theory. Second, process evaluations should aim to understand interventions’ mechanism of action, rather than simply ‘what happened’. Third, small scoping studies should be conducted in new settings, to explore how to enact identified mechanisms. Finally, innovative synthesis methods are required, in order to identify mechanisms of action where there is a lack of existing process evaluations.