bluuebunny/crossref_metadata_2025 huggingface dataset:
Policy levers and priority-setting in universal health coverage: a qualitative analysis of healthcare financing agenda setting in Kenya
Authors: Oraro-Lawrence Tessa , Wyss Kaspar
Published: March 2020

Background

Competing priorities in health systems necessitate difficult choices on which health actions and investments to fund: decisions that are complex, value-based, and highly political. In light of the centrality of universal health coverage (UHC) in driving current health policy, we sought to examine the value interests that influence agenda setting in the country’s health financing space. Given the plurality of Kenya’s health policy levers, we aimed to examine how the perspectives of stakeholders involved in policy decision-making and implementation shape discussions on health financing within the UHC framework.

Methods

A series of in-depth key informant interviews were conducted at national and county level (n = 13) between April and May 2018. Final thematic analysis using the Framework Method was conducted to identify similarities and differences amongst stakeholders on the challenges hindering Kenya’s achievement of UHC in terms of its the optimisation of health service coverage; expansion of the population that benefits from essential healthcare services; and the minimisation of out-of-pocket costs associated with health-seeking behaviour.

Results

Our findings indicate that the perceived lack of strategic leadership from Kenya’s national government has led to a lack of agreement on stakeholders’ interpretation of what is to be understood by UHC, its contextual values and priorities. We observe material differences between and within policy networks on the country’s priorities for population coverage, healthcare service provision, and cost-sharing under the UHC dispensation. In spite of this, we note that progressive universalism is considered as the preferred approach towards UHC in Kenya, with most interviewees prioritising an equity-based approach that prioritises better access to healthcare services and financial risk protection. However, the conflicting priorities of key stakeholders risk derailing progress towards the expansion of access to health services and financial risk protection.

Conclusions

This study adds to existing knowledge of UHC in Kenya by contextualising the competing and evolving priorities that should be taken into consideration as the country strategises over its UHC process. We suggest that clear policy action is required from national government and county governments in order to develop a logical and consistent approach towards UHC in Kenya.
LncRNA MALAT1 silencing protects against cerebral ischemia-reperfusion injury through miR-145 to regulate AQP4
Authors: Wang Hongwei , Zheng Xiaoxiao , Jin Jing , Zheng Li , Guan Ting , Huo Yangfan , Xie Shufen , Wu Ying , Chen Wei
Published: March 2020

Background

The present study aimed to verify whether long noncoding RNA (lncRNA) MALAT1 is involved in brain tissue damage induced by ischemia-reperfusion injury, and to explore the mechanism by which MALAT1 regulates aquaporin 4 (AQP4).

Methods

In this study, we established glucose deprivation (OGD)/reoxygenation (RX) astrocyte cell model and middle cerebral artery occlusion (MCAO)/reperfusion mouse model in vitro and in vivo. Then cell counting kit-8 assay, flow cytometry analysis, Triphenyltetrazolium chloride (TTC) staining, and western blotting were used to determine cell viability, cell apoptosis, cerebral infarction volume, and the abundance of AQP4, respectively.

Results

We found that the level of MALAT1 was significantly upregulated in both the MCAO/reperfusion model and OGD/RX model. Knockdown of MALAT1 increased cell viability and reduced cell apoptosis in MA-C cells, while an AQP4 siRNA combined with a siRNA targeting MALAT1 could not enhance this effect. Further experiments showed that MALAT1 positively regulated AQP4 expression via miR-145. The MALAT1 siRNA did not alleviate the exacerbation of damage after miR-145 inhibitor action. However, an miR-145 inhibitor reversed the protection effects of MALAT1, indicating that MALAT1 silencing protects against cerebral ischemia-reperfusion injury through miR-145. TTC staining showed that the infracted area of whole brain was significantly attenuated in treated with sh-MALAT1 group in vivo.

Conclusion

Taken together, our study confirmed that MALAT1 promotes cerebral ischemia-reperfusion injury by affecting AQP4 expression through competitively binding miR-145, indicating that MALAT1 might be a new therapeutic target for treatment cerebral ischemic stroke.
Optimized expression and enhanced production of alkaline protease by genetically modified Bacillus licheniformis 2709
Authors: Zhou Cuixia , Zhou Huiying , Li Dengke , Zhang Huitu , Wang Hongbin , Lu Fuping
Published: March 2020

Background

Bacillus licheniformis2709 is extensively applied as a host for the high-level production of heterologous proteins, butBacilluscells often possess unfavorable wild-type properties, such as production of viscous materials and foam during fermentation, which seriously influenced the application in industrial fermentation. How to develop it from a soil bacterium to a super-secreting cell factory harboring less undomesticated properties always plays vital role in industrial production. Besides, the optimal expression pattern of the inducible enzymes like alkaline protease has not been optimized by comparing the transcriptional efficiency of different plasmids and genomic integration sites inB. licheniformis.

Result

Bacillus licheniformis2709 was genetically modified by disrupting the nativelchACgenes related to foaming and theepscluster encoding the extracellular mucopolysaccharide via a markerless genome-editing method. We further optimized the expression of the alkaline protease gene (aprE) by screening the most efficient expression system among different modular plasmids and genomic loci. The results indicated that genomic expression ofaprEwas superior to plasmid expression and finally the transcriptional level ofaprE greatly increased 1.67-fold through host optimization and chromosomal integration in the vicinity of the origin of replication, while the enzyme activity significantly improved 62.19% compared with the wild-type alkaline protease-producing strainB. licheniformis.

Conclusion

We successfully engineered an AprE high-yielding strain free of undesirable properties and its fermentation traits could be applied to bulk-production by host genetic modification and expression optimization. In summary, host optimization is an enabling technology for improving enzyme production by eliminating the harmful traits of the host and optimizing expression patterns. We believe that these strategies can be applied to improve heterologous protein expression in otherBacillusspecies.
Long non-coding RNASNHG17 promotes the progression of breast cancer by sponging miR-124-3p
Authors: Du Ye , Wei Na , Hong Jinghui , Pan Weiyun
Published: March 2020

Background

Small nucleolar RNA host gene 17 (SNHG17), a novel cancer-related long noncoding RNA (lncRNA), was reported to be responsible for processing and developing in several cancers. Nonetheless, the clinical significance and biological function of SNHG17 in human breast cancer (BC) remain rarely known.

Materials and methods

58 pairs of BC tissues and adjacent non-cancerous tissues were harvested to measure SNHG17 expression levels. SNHG17 was knockdown to study its biological behavior in BC cells. The microRNAs (miRNAs) that can bind to SNHG17 were predicated using Starbase2.0 and were tested using luciferase reporter activity and RIP assays. A xenograft model was established to investigate the impact of SNHG17 in tumor growth in vivo.

Results

An increased SNHG17 was observed in BC samples and cell lines compared with corresponding control. Increased SNHG17 was closely associated with poor prognosis.SNHG17 depletion suppressed cell proliferation, migration and invasion in vitro, as well as inhibited tumor growth in xenograft tumor models. Mechanistically, SNHG17 could function as an endogenous sponge of miR-124-3p in BC cells. Moreover, the repression of cell proliferation, migration and invasion induced by SNHG17 knockdown would reversed by miR-124-3p inhibitor.

Conclusion

The present study demonstrated that the lncRNASNHG17 could regulate the progression of BC by sponging miR-124-3p.
The clinical prognostic value of lncRNA FAM83H-AS1 in cancer patients: a meta-analysis
Authors: Yang Qin , Wang Jie , Zhong Pingyong , Mou Tinggang , Hua Hao , Liu Pan , Xie Fei
Published: March 2020

Background

Family with sequence similarity 83 member H antisense RNA 1 (FAM83H-AS1) is a novel long non-coding RNA. Increasing studies have reported that FAM83H-AS1 is abnormally expressed in a variety of tumors and is associated with poor outcome. However, the clinical prognostic significance of lncRNA FAM83H-AS1 in tumors is not completely known.

Methods

In this meta-analysis, literature was collected up until February 5, 2020 through multifarious retrieval strategies by searching through electronic databases of PubMed, Cochrane Library, EMBASE, Medline, Web of Science, CNKI, Weipu, and Wanfang. A total of 14 studies that met the inclusion criteria with relevant clinical data and prognostic information were included in the meta-analysis.

Results

The combined results revealed that high expression of FAM83H-AS1 was associated with poor overall survival (OS) (HR = 1.63, 95% CI 1.24–2.14, P = 0.0004) in a variety of cancers. Additionally, upregulated FAM83H-AS1 expression was significantly correlated with tumor TNM stage (III/IV vs. I/II, OR = 2.40, 95% CI 1.36–4.23, P = 0.003) and lymph node metastasis (positive vs. negative, OR = 1.70, 95% CI 1.14–2.52, P = 0.008) in patients with cancer.

Conclusions

Our results of this meta-analysis indicated that elevated FAM83H-AS1 expression could predict poor prognosis in patients with cancer and suggested that FAM83H-AS1 might serve as a novel biomarker for cancer.
Effects of dynamic radial tensile stress on fibrocartilage differentiation of bone marrow mesenchymal stem cells
Authors: Su Xuelian , Wang Jizeng , Kang Hong , Bao Guangjie , Liu Lin
Published: March 2020

Background

Uniaxial/biaxial tensile stress has been employed to induce chondrocyte differentiation of mesenchymal stem cells. However, the effects of radial tensile stimuli on differentiation of MSCs into fibrocartilage remain unclear.

Results

It was found that induced bone marrow mesenchymal stem cells (BMSCs) were not only similar to TMJ disc cells in morphology, but also could synthesize type I collagen (Col I), a small amount of type II collagen (Col II) and glycosaminoglycans (GAGs). The synthesis of Col I significantly increased while that of Col II gradually decreased with increasing tensile strength. The ratio of Col I to Col II was 1.8 to 1 and 2 to 1 in the 10% and 15% stretching groups, respectively. The gene expression of Col I and GAGs was significantly upregulated, whereas that of Col II was downregulated. However, the higher tensile stimulation (15%) promoted the synthesis of α-smooth muscle actin (α-SMA). Too much α-SMA is not conducive to constructing engineered tissue.

Conclusion

Therefore, the 10% radial tensile stimulus was the optimal strength for inducing the BMSCs to differentiate into fibrochondrocytes of the temporomandibular joint (TMJ) disc. This work provided a novel approach for inducing BMSCs to differentiate into fibrochondrocytes.
A new immune signature for survival prediction and immune checkpoint molecules in lung adenocarcinoma
Authors: Guo Dina , Wang Mian , Shen Zhihong , Zhu Jiaona
Published: March 2020

Background

Lung adenocarcinoma (LUAD) is the most frequent subtype of lung cancer. The prognostic signature could be reliable to stratify LUAD patients according to risk, which helps the management of the systematic treatments. In this study, a systematic and reliable immune signature was performed to estimate the prognostic stratification in LUAD.

Methods

The profiles of immune-related genes for patients with LUAD were used as one TCGA training set: n = 494, other validation set 1: n = 226 and validation set 2: n = 398. Univariate Cox survival analysis was used to identify the candidate immune-related genes from each cohort. Then, the immune signature was developed and validated in the training and validation sets.

Results

In this study, functional analysis showed that immune-related genes involved in immune regulation and MAPK signaling pathway. A prognostic signature based on 10 immune-related genes was established in the training set and patients were divided into high-risk and low-risk groups. Our 10 immune-related gene signature was significantly related to worse survival, especially during early-stage tumors. Further stratification analyses revealed that this 10 immune-related gene signature was still an effective tool for predicting prognosis in smoking or nonsmoking patients, patients with KRAS mutation or KRAS wild-type, and patients with EGFR mutation or EGFR wild-type. Our signature was negatively correlated with B cell, CD4+ T cell, CD8+ T cell, neutrophil, dendritic cell (DC), and macrophage immune infiltration, and immune checkpoint molecules PD-1 and CTLA-4 (P < 0.05).

Conclusions

These findings suggested that our signature was a promising biomarker for prognosis prediction and can facilitate the management of immunotherapy in LUAD.
Health-related quality of life in hereditary transthyretin amyloidosis polyneuropathy: a prospective, observational study
Authors: Inês Mónica , Coelho Teresa , Conceição Isabel , Ferreira Lara , de Carvalho Mamede , Costa João
Published: March 2020

Background

Hereditary Transthyretin Amyloidosis Polyneuropathy is a rare life-threatening neurologic disease that imposes considerable mortality and it is associated with progressive related disabilities. In this study, we aimed to assess the effect of the disease across health-related quality of life dimensions, in both carriers of the mutation and patients, to compare health-related quality of life with general population, as well as to explore health-related quality of life prognostic factors among patients, including disease progression and treatment.

Methods

This study was a multi-institutional, longitudinal, prospective, observational study of hereditary Transthyretin Amyloidosis Polyneuropathy Portuguese adult subjects (621 asymptomatic carriers and 733 symptomatic patients) enrolled in the Transthyretin Amyloidosis Outcomes Survey. Health-related quality of life was captured with the preference-based instrument EQ-5D-3 L. For general population the dataset included all subjects enrolled in a representative national study (n = 1500). Different econometric models were specified; multivariate probit, generalized linear model and generalized estimating equations model; including demographic and clinical covariates.

Results

Hereditary Transthyretin Amyloidosis Polyneuropathy patients have their health status severely impaired in all quality of life dimensions and more anxiety/depression problems were found among asymptomatic carriers. No differences on utility were found between carriers and general population (p = 0.209). Among patients, the utility value is estimated to be 0.51 (0.021), a decrement of 0.27 as compared with general population utility. Higher disease duration, advanced disease stage and not receiving treatment are associated with impaired health-related quality of life. No differences were found between genders (p = 0.910) or between late (≥50 years) and early-onset patients (p = 0.254). The utility estimate ranged from 0.63 (0.009) in stage I to 0.01 (0.005) in stage IV.

Conclusions

Hereditary Transthyretin Amyloidosis Polyneuropathy symptoms and progressive associated disabilities substantially decrease patient’s health-related quality of life. Clinical strategies focused on health-related quality of life preservation such as close follow-up of asymptomatic carriers, prompt diagnosis and adequate, early treatment would benefit patient’s long-term outcomes, slowing the progressive decline in health-related quality of life.
Understanding acute metabolic decompensation in propionic and methylmalonic acidemias: a deep metabolic phenotyping approach
Authors: Haijes H. A. , Jans J. J. M. , van der Ham M. , van Hasselt P. M. , Verhoeven-Duif N. M.
Published: March 2020

Background

Pathophysiology of life-threatening acute metabolic decompensations (AMD) in propionic acidemia (PA) and isolated methylmalonic acidemia (MMA) is insufficiently understood. Here, we study the metabolomes of PA and MMA patients over time, to improve insight in which biochemical processes are at play during AMD.

Methods

Longitudinal data from clinical chemistry analyses and metabolic assays over the life-course of 11 PA and 13 MMA patients were studied retrospectively. Direct-infusion high-resolution mass spectrometry was performed on 234 and 154 remnant dried blood spot and plasma samples of PA and MMA patients, respectively. In addition, a systematic literature search was performed on reported biomarkers. All results were integrated in an assessment of biochemical processes at play during AMD.

Results

We confirmed many of the metabolite alterations reported in literature, including increases of plasma valine and isoleucine during AMD in PA patients. We revealed that plasma leucine and phenylalanine, and urinary pyruvic acid were increased during AMD in PA patients. 3-hydroxyisovaleric acid correlated positively with plasma ammonia. We found that known diagnostic biomarkers were not significantly further increased, while intermediates of the branched-chain amino acid (BCAA) degradation pathway were significantly increased during AMD.

Conclusions

We revealed that during AMD in PA and MMA, BCAA and BCAA intermediates accumulate, while known diagnostic biomarkers remain essentially unaltered. This implies that these acidic BCAA intermediates are responsible for metabolic acidosis. Based on this, we suggest to measure plasma 3-hydroxyisovaleric acid and urinary ketones or 3-hydroxybutyric acid for the biochemical follow-up of a patient’s metabolic stability.
Sex differences in blood pro-oxidant status and platelet activation in children admitted with respiratory syncytial virus bronchiolitis: a pilot study
Authors: De Jacobis Isabella Tarissi , Vona Rosa , Straface Elisabetta , Gambardella Lucrezia , Ceglie Giulia , de Gennaro Francesca , Pontini Ilenia , Vittucci Anna Chiara , Carè Alessandra , Cittadini Camilla , Villani Alberto , Pietraforte Donatella
Published: March 2020

Background

Respiratory syncytial virus (RSV) is the most common cause of bronchiolitis in the pediatric population worldwide and an important cause of death in developing countries. It has been demonstrated that the balance between oxidant and antioxidant systems is disrupted in children with bronchiolitis and that oxidative stress contributes to the pathogenesis of this disease. Platelets play an important role in antimicrobial host defenses and contribute to pulmonary vascular repair being either targets or source of reactive oxidizing species. The main purpose of this study was to assessing sex differences in clinical characteristics and platelets activation during RSV bronchiolitis in infancy.

Methods

In this retrospective study a total of 203 patients (112 boys and 91 girls) with bronchiolitis, aged 12 months or less, admitted to the Bambino Gesù Pediatric Hospital of Rome (Italy) in the period from January to December 2017, were enrolled. Moreover, in a select group of patients (15 boys and 12 girls) with diagnosis of moderate bronchiolitis from RSV, a pilot study on oxidative stress and platelet characteristics was carried out by electron paramagnetic resonance and flow cytometry respectively. Age-matched healthy control subjects (10 boys and 10 girls) were chosen as controls. Data were analyzed using Student’ T test, Chi Squared test and one-way ANOVA test.

Results

This study highlights the influence of sex in the clinical course of bronchiolitis. In particular we found: i) a higher incidence of bronchiolitis in boys than in girls (55% vs 45%); ii) higher C reactive protein values in girls than boys (1.11 mg/dL vs 0.92 mg/dL respectively; p < 0.05); iii) a different degree of thrombocytosis during hospitalization (mild in the girls and severe in the boys). Moreover, in selected patients we found that compared to girls with bronchiolitis, boys showed: i) higher percentage of activated platelets (8% vs 2% respectively; p < 0.05) and iii) higher number of platelets forming homotypic aggregates (2.36% vs 0.84% respectively, p < 0.05).

Conclusion

The present study affirm that the bronchiolitis is an infection in which sex seems to act as a modulating factor only in the clinical course, influencing also the choice of the therapy should be made.
Comparison of infectivity of Plasmodium vivax to wild-caught and laboratory-adapted (colonized) Anopheles arabiensis mosquitoes in Ethiopia
Authors: Chali Wakweya , Ashine Temesgen , Hailemeskel Elifaged , Gashaw Abrham , Tafesse Temesgen , Lanke Kjerstin , Esayas Endashaw , Kedir Soriya , Shumie Girma , Behaksra Sinknesh Wolde , Bradley John , Yewhalaw Delenasaw , Mamo Hassen , Petros Beyene , Drakeley Chris , Gadisa Endalamaw , Bousema Teun , Tadesse Fitsum G.
Published: March 2020

Background

Mosquito-feeding assays that assess transmission of Plasmodium from man-to-mosquito typically use laboratory mosquito colonies. The microbiome and genetic background of local mosquitoes may be different and influence Plasmodium transmission efficiency. In order to interpret transmission studies to the local epidemiology, it is therefore crucial to understand the relationship between infectivity in laboratory-adapted and local mosquitoes.

Methods

We assessed infectivity of Plasmodium vivax-infected patients from Adama, Ethiopia, using laboratory-adapted (colony) and wild-caught (wild) mosquitoes raised from larval collections in paired feeding experiments. Feeding assays used 4–6 day-old female Anopheles arabiensis mosquitoes after starvation for 12 h (colony) and 18 h (wild). Oocyst development was assessed microscopically 7 days post-feeding. Wild mosquitoes were identified morphologically and confirmed by genotyping. Asexual parasites and gametocytes were quantified in donor blood by microscopy.

Results

In 36 paired experiments (25 P. vivax infections and 11 co-infections with P. falciparum), feeding efficiency was higher in colony (median: 62.5%; interquartile range, IQR: 47.0–79.0%) compared to wild mosquitoes (median: 27.8%; IQR: 17.0–38.0%; Z = 5.02; P < 0.001). Plasmodium vivax from infectious individuals (51.6%, 16/31) infected a median of 55.0% (IQR: 6.7–85.7%; range: 5.5–96.7%; n = 14) of the colony and 52.7% (IQR: 20.0–80.0%; range: 3.2–95.0%; n = 14) of the wild mosquitoes. A strong association (ρ(16) = 0.819; P < 0.001) was observed between the proportion of infected wild and colony mosquitoes. A positive association was detected between microscopically detected gametocytes and the proportion of infected colony (ρ(31) = 0.452; P = 0.011) and wild (ρ(31) = 0.386; P = 0.032) mosquitoes.

Conclusions

Infectivity assessments with colony and wild mosquitoes yielded similar infection results. This finding supports the use of colony mosquitoes for assessments of the infectious reservoir for malaria in this setting whilst acknowledging the importance of mosquito factors influencing sporogonic development of Plasmodium parasites.
Genetic predisposition to smoking is associated with risk of rheumatoid arthritis: a Mendelian randomization study
Authors: Qian Yu , Zhang Lingzhi , Wu David J. H. , Xie Zhijun , Wen Chengping , Mao Yingying
Published: March 2020

Background

Although observational epidemiological studies have found that smoking is positively associated with risk of rheumatoid arthritis (RA), assessing the causality of this relationship has remained elusive because conventional observational studies are susceptible to bias such as confounding and reverse causation. Here, we applied the Mendelian randomization (MR) approach to examine the potential causal relationship between smoking and risk of RA.

Methods

Summary statistics data for RA were obtained from a meta-analysis of genome-wide association studies (GWAS), including 14,361 RA cases and 43,923 controls of European ancestry. The instrumental variables (IV) and the genetic association estimates for smoking initiation and lifetime smoking were obtained from a GWAS meta-analysis including 1,232,091 individuals and a GWAS of 462,690 individuals of European ancestry, respectively. MR analyses were performed using the inverse-variance weighted (IVW) method and supplemented with the weighted-median method. Potential pleiotropy was assessed using the MR-Pleiotropy RESidual Sum and Outlier (MR-PRESSO) test and MR-Egger regression. Sensitivity analyses were further performed to test the robustness of the association.

Results

We found that compared with never smokers, genetic predisposition to smoking initiation was positively associated with risk of RA (odds ratio (OR) = 1.32, 95% confidence interval (CI) = 1.15–1.52, P = 9.17 × 10−5 using the IVW method). Similarly, genetically predicted lifetime smoking was associated with an increased risk of RA (OR = 1.55, 95% CI = 1.13–2.14, P = 0.007). Sensitivity analyses using alternative MR methods and different sets of IVs produced similar results, suggesting the robustness of our findings.

Conclusions

These results provide support for a causal association between smoking and increased risk of RA. Further studies are warranted to explain the underlying mechanisms of smoking in the development of RA.
Thoracic ultrasound evaluation and B-type natriuretic peptide value in elective cesarean section under spinal anesthesia
Authors: Vetrugno Luigi , Dogareschi Teresa , Sassanelli Rossella , Orso Daniele , Seremet Ludmilla , Mattuzzi Lisa , Scapol Sara , Spasiano Alessandra , Cagnacci Angelo , Bove Tiziana
Published: March 2020

Background

Pregnancy-induced changes in cardiovascular status make women more susceptible to pulmonary edema. During cesarean section, to counterbalance the effect of hypotension caused by spinal anesthesia, anesthesiologists must choose between two fundamental approaches to maintain the hemodynamic state—intravenous fluids or vasopressors—and this choice will depend upon their particular opinions and experience. We aim to assess for any correlations between thoracic ultrasound A- and B-line artifacts, brain natriuretic peptide (BNP) levels, and the amount of intraoperative fluids administered.

Results

From December 2016 to August 2018, at the University-Hospital of Udine, we enrolled 80 consecutive pregnant women undergoing cesarean section. We observed a statistically significant difference in the volume of fluids administered in the first 24 h (p = 0.035) between the patients presenting B-lines in at least one basal area of their thoracic ultrasound and patients with no evident B-lines (AUC 66.4%; IC 0.49–0.83). Dividing the population on whether their BNP levels were higher or less than 20 pg/mL, no statistically significant difference was revealed with regard to fluids administered in the first 24 h (p = 0.537).

Conclusions

Thoracic ultrasound is a non-invasive and easy-to-use tool for detecting fluid intolerance in pregnant women undergoing cesarean section. BNP levels were slow to rise following the cesarean section and did not show any clear correlation with fluid volumes administered.
Determining spa-type of methicillin-resistant Staphylococcus aureus (MRSA) via high-resolution melting (HRM) analysis, Shiraz, Iran
Authors: Hashemizadeh Zahra , Bazargani Abdollah , Kalantar-Neyestanaki Davood , Mohebi Samane , Hadi Nahal
Published: March 2020

Objectives

Molecular typing methods are useful for rapid detection and control of a disease. Recently, the use of high-resolution melting (HRM) for spa typing of MRSA isolates were reported. This technique is rapid, inexpensive and simple for genotyping and mutation screening in DNA sequence. The aim of this study was to evaluate the ability of HRM-PCR to analysis spa genes amongst MRSA isolates.

Results

A total of 50 MRSA isolates were collected from two teaching hospitals in Shiraz, Iran. The isolates were confirmed as MRSA by susceptibility to cefoxitin and detection of mecA gene using PCR. We used HRM analysis and PCR-sequencing method for spa typing of MRSA isolates. In total, 15 different spa types were discriminate by HRM and sequencing method. The melting temperature of the 15 spa types, using HRM genotyping were between 82.16 and 85.66 °C. The rate of GC % content was 39.4–46.3. According to the results, spa typing of 50 clinical isolates via PCR-sequencing and HRM methods were 100% similar. Consequently, HRM method can easily identify and rapidly differentiate alleles of spa genes. This method is faster, less laborious and more suitable for high sample at lower cost and risk of contamination.
Exploratory analysis of myocardial function after extracorporeal cardiopulmonary resuscitation vs conventional cardiopulmonary resuscitation
Authors: Tonna Joseph E. , McKellar Stephen H. , Selzman Craig H. , Drakos Stavros , Koliopoulou Antigone G. , Taleb Iosif , Stoddard Gregory J. , Stehlik Josef , Welt Frederick G. P. , Fair James F. , Stoddard Kathleen , Youngquist Scott T.
Published: March 2020

Objective

Ventricular unloading is associated with myocardial recovery. We sought to evaluate the association of extracorporeal cardiopulmonary resuscitation (ECPR) on myocardial function after cardiac arrest. We conducted a retrospective exploratory analysis, comparing ejection fraction (EF) after adult cardiac arrest, between ECPR and conventional CPR.

Results

Among 1119 cases of cardiac arrest, 116 had an echocardiogram post-return of spontaneous circulation (ROSC) and were included. Thirty-eight patients had ≥ 2 echocardiograms. ECPR patients had differences in age, hypertension and chronic heart failure. ECPR patients had a lower EF post-ROSC (24% vs 45%;p < 0.01) and were more likely to undergo percutaneous coronary intervention (25% vs 3%; p < 0.01). In multivariate analysis, only ECPR use (β-coeff: 10.4 [95% CI 3.68–17.13];p < 0.01) independently predicted improved myocardial function. In this exploratory study, EF after cardiac arrest may be more likely to improve among ECPR patients than CCPR patients. Our methodology should be replicated to confirm or refute the validity of our findings.
Clinical application of multi-material artifact reduction (MMAR) technique in Revolution CT to reduce metallic dental artifacts
Authors: Wei Yijuan , Jia Fei , Hou Ping , Zha Kaiji , Pu Shi , Gao Jianbo
Published: March 2020

Background

This study aimed to explore the performance of Revolution CT virtual monoenergetic images (VMI) combined with the multi-material artifact reduction (MMAR) technique in reducing metal artifacts in oral and maxillofacial imaging.

Results

There were significant differences in image quality scores between VMI + MMAR images and VMI+MARS (multiple artifact reduction system) images at each monochromatic energy level (p = 0.000). Compared with the MARS technology, the MMAR technology further reduced metal artifacts and improved the image quality. At VMI90 keV and VMI110 keV, the SD, CNR, and AI in the Revolution CT group were significantly lower than in the Discovery CT, but no significant differences in these parameters were found between two groups at VMI50 keV, VMI70 keV, and VMI130 keV (p > 0.05). The attenuation was comparable between two groups at any energy level (p > 0.05).

Conclusions

Compared with the MARS reconstruction technique of Discovery CT, the MMAR technique of Revolution CT is better to reduce the artifacts of dental implants in oral and maxillofacial imaging, which improves the image quality and the diagnostic value of surrounding soft tissues.
Identification of the initial molecular changes in response to circulating angiogenic cells-mediated therapy in critical limb ischemia
Authors: Beltran-Camacho Lucia , Jimenez-Palomares Margarita , Rojas-Torres Marta , Sanchez-Gomar Ismael , Rosal-Vela Antonio , Eslava-Alcon Sara , Perez-Segura Mª Carmen , Serrano Ana , Antequera-González Borja , Alonso-Piñero Jose Angel , González-Rovira Almudena , Extremera-García Mª Jesús , Rodriguez-Piñero Manuel , Moreno-Luna Rafael , Larsen Martin Røssel , Durán-Ruiz Mª Carmen
Published: March 2020

Background

Critical limb ischemia (CLI) constitutes the most aggressive form of peripheral arterial occlusive disease, characterized by the blockade of arteries supplying blood to the lower extremities, significantly diminishing oxygen and nutrient supply. CLI patients usually undergo amputation of fingers, feet, or extremities, with a high risk of mortality due to associated comorbidities.

Circulating angiogenic cells (CACs), also known as early endothelial progenitor cells, constitute promising candidates for cell therapy in CLI due to their assigned vascular regenerative properties. Preclinical and clinical assays with CACs have shown promising results. A better understanding of how these cells participate in vascular regeneration would significantly help to potentiate their role in revascularization.

Herein, we analyzed the initial molecular mechanisms triggered by human CACs after being administered to a murine model of CLI, in order to understand how these cells promote angiogenesis within the ischemic tissues.

Methods

Balb-c nude mice (n:24) were distributed in four different groups: healthy controls (C, n:4), shams (SH, n:4), and ischemic mice (after femoral ligation) that received either 50 μl physiological serum (SC, n:8) or 5 × 105 human CACs (SE, n:8). Ischemic mice were sacrificed on days 2 and 4 (n:4/group/day), and immunohistochemistry assays and qPCR amplification of Alu-human-specific sequences were carried out for cell detection and vascular density measurements. Additionally, a label-free MS-based quantitative approach was performed to identify protein changes related.

Results

Administration of CACs induced in the ischemic tissues an increase in the number of blood vessels as well as the diameter size compared to ischemic, non-treated mice, although the number of CACs decreased within time. The initial protein changes taking place in response to ischemia and more importantly, right after administration of CACs to CLI mice, are shown.

Conclusions

Our results indicate that CACs migrate to the injured area; moreover, they trigger protein changes correlated with cell migration, cell death, angiogenesis, and arteriogenesis in the host. These changes indicate that CACs promote from the beginning an increase in the number of vessels as well as the development of an appropriate vascular network.

Graphical abstract
Results from extended lymphadenectomies with [111In]PSMA-617 for intraoperative detection of PSMA-PET/CT-positive nodal metastatic prostate cancer
Authors: Jilg Cordula A. , Reichel Kathrin , Stoykow Christian , Rischke H. Christian , Bartholomä Mark , Drendel Vanessa , von Büren Moritz , Schultze-Seemann Wolfgang , Meyer Philipp T. , Mix Michael
Published: March 2020

Purpose

Identification of suspicious PSMA-PET/CT-positive lymph node (LN) metastases (LNM) from prostate cancer (PCa) during lymphadenectomy (LA) is challenging. We evaluated an 111In-labelled PSMA ligand (DKFZ-617, referred to as [111In]PSMA-617) as a γ-emitting tracer for intraoperative γ-probe application for resected tissue samples in PCa patients. Forty-eight hours prior to LA, [111In]PSMA-617 was administered intravenously in 23 patients with suspected LNM on PSMA-PET/CT (n = 21 with biochemical relapse, n = 2 at primary therapy). Resected tissue samples (LN, LNM and fibrofatty tissue) were measured ex situ by a γ-probe expressed as counts per second (CPSnorm). [111In]PSMA-617 tissue sample uptake was measured by a germanium detector for verification and calculated as %IAlbm (percent injected activity per kilogram lean body mass at time of surgery). Based on a clinical requirement for a specificity > 95%, thresholds for both ex situ measurements were chosen accordingly. Correlation of the results from PET/CT, γ-probe and germanium detector with histopathology was done.

Results

Eight hundred sixty-four LNs (197 LNM) were removed from 275 subregions in 23 patients, on average 8.6 ± 14.9 LNM per patient. One hundred four of 275 tissue samples showed cancer. Median γ-probe and germanium detector results were significantly different between tumour-affected (33.5 CPSnorm, 0.71 %IAlbm) and tumour-free subregions (3.0 CPSnorm, 0.03 %IAlbm) (each p value < 0.0001). For the chosen γ-probe cut-off (CPSnorm > 23) and germanium detector cut-off (%IAlbm > 0.27), 64 and 74 true-positive and 158 true-negative samples for both measurements were identified. Thirty-nine and 30 false-negative and 6 and 5 false-positive tissue samples were identified by γ-probe and germanium detector measurements.

Conclusion

[111In]PSMA-617 application for LA is feasible in terms of an intraoperative real-time measurement with a γ-probe for detection of tumour-affected tissue samples. γ-probe results can be confirmed by precise germanium detector measurements and were significantly different between tumour-affected and tumour-free samples.
The equivalence of $F_{a}$-frames
Authors: Hussain Tufail , Li Yun-Zhang
Published: March 2020

Structured frames such as wavelet and Gabor frames in$L^{2}(\mathbb {R})$L2(R)have been extensively studied. But$L^{2}(\mathbb{ R}_{+})$L2(R+)cannot admit wavelet and Gabor systems due to$\mathbb{R}_{+}$R+being not a group under addition. In practice,$L^{2}(\mathbb{R}_{+})$L2(R+)models the causal signal space. The function-valued inner product-based$F_{a}$Fa-frame for$L^{2}(\mathbb{R}_{+})$L2(R+)was first introduced by Hasankhani Fard and Dehghan, where an$F_{a}$Fa-frame was called a function-valued frame. In this paper, we introduce the notions of$F_{a}$Fa-equivalence and unitary$F_{a}$Fa-equivalence between$F_{a}$Fa-frames, and present a characterization of the$F_{a}$Fa-equivalence and unitary$F_{a}$Fa-equivalence. This characterization looks like that of equivalence and unitary equivalence between frames, but the proof is nontrivial due to the particularity of$F_{a}$Fa-frames.
Dynamics and oscillations of models for differential equations with delays
Authors: Miraoui Mohsen , Repovš Dušan D.
Published: March 2020

By developing new efficient techniques and using an appropriate fixed point theorem, we derive several new sufficient conditions for the pseudo almost periodic solutions with double measure for some system of differential equations with delays. As an application, we consider certain models for neural networks with delays.
Trends in survival of stage IV breast cancer (BC) among Caucasian and African American (AA) BC patients (pts)
Authors: Dawood S. S. , Broglio K. , Hortobagyi G. , Giordano S.
Published: March 2020

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Background: The aim of this review was to look at the trends in survival over time in patients(pts) with de novo stage IV BC and to identify factors that impact survival. Methods: We searched the SEER registry to identify pts with de novo stage IV breast cancer diagnosed between 1988 and 2003. Pts who were male, did not have microscopic confirmation of disease, had more than one primary, and who had a prior malignancy were excluded. Pts were divided into three groups based on their year of diagnosis separated into 5-year intervals: 1988 - 1993, 1994 - 1998, and 1999 -2003. Overall survival (OS) and BC specific survival (BCS) were calculated from the date of diagnosis to the date of death, last follow-up, or 12/31/03. Survival outcomes were estimated by the Kaplan-Meier method, and Cox models were fit to determine the characteristics that were independently associated with survival. Follow-up time was censored at 3 years for Cox analyses. Results: 15,438 pts were identified. 3796 pts were diagnosed from 1988–1993, 3954 from 1994–1998, and 7688 from 1999–2003. Median age was 62 years. Median follow-up was 16 months (mos) (range 0–191), 18 mos (range 0–199), and 11 mos (range 0–59) in periods 1988 - 1993, 1994 - 1998, and 1999 - 2003 respectively. Median OS was 18 mos overall and 16, 18, and 20 mos respectively for each time period. Median BCS was 23 mos overall and 20, 21, and 25 mos for each time period. In the multivariable model, more distant year of diagnosis, Grade 3 disease, higher number of positive LN, increasing age, being unmarried, ER- disease, PR- disease, and no surgery were all independently associated with worse BCS. The interaction term between ER and year of diagnosis was of borderline significance, indicating that over time, pts with ER+ disease had decreasing risk of death compared to patients with ER- disease (HR 0.98, 95% CI 0.96–1.00). An interaction term between race and year of diagnosis was significant, such that with each increasing year of diagnosis AA pts had increasing risk of death compared to whites (HR 1.04, 95% CI 1.01–1.07). Conclusions: The survival of de novo stage IV BC pts has modestly improved over time. Of concern, survival disparities between AA and white patients increased rather than diminished over time.

No significant financial relationships to disclose.
Sensitivity of myxoid-round cell liposarcoma (MRCL) to trabectedin (T) may be related to a direct effect on the fusion transcript
Authors: Grosso F. , Forni C. , Frapolli R. , Greco A. , Gronchi A. , Jimeno J. , Mantovani R. , D'Incalci M. , Pilotti S. , Casali P. G.
Published: March 2020

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Background: Myxoid-round cell liposarcomas (MRCL) is a variant of liposarcoma, associated with t(12;16)(q13;p11) and the rarer t(12;22)(q13;q12) chromosomal translocation, resulting in the FUS-CHOP and EWS-CHOP fusion proteins, supposed to act as aberrant transcription factors. We reported the exceedingly high clinical efficacy of T against MRCL, which is a magnitude higher in comparison to other liposarcomas and sarcomas. Reasons thereof remain unknown, but a distinct mechanism of action of T in MRCL is suspected on clinical grounds. Methods: Tumor biopsies were taken from 9 MRCL patients (pts) before starting T, and the tumor was fully characterized by cytogenetic and molecular analysis. Primary cultures were set up, and tumor fragments were transplanted in scid mice, to develop an experimental model aimed at investigating the mechanism of action of T. Chromatin immunoprecipitation (ChIP) analysis on the 402–91 myxoid liposarcoma cell line was performed, to characterize the DNA binding capability of FUS-CHOP. Finally, in vivo ChIP analyses are currently ongoing, using tumor samples obtained before and after therapy with T. Tumor samples from 3 patients have been processed so far. Results: All patients had t(12–16) by FISH. Molecular characterization showed that 7 pts had the type II transcript, 1 had the type III, and 1 both type II and III. Primary cultures evaluated after the first 3 passages were morphologically, cytogenetically and molecularly consistent with the original tumor, as well as the 4 tumours grown in mice so far. In the myxoid liposarcoma cell line carrying the type I transcript, selective binding of FUS-CHOP to some promoters was seen. Impairment of this binding activity was observed after treatment with T at concentration 1–4 nM. Conclusions: There is in vitro evidence of a direct effect of T on the fusion protein's ability to bind to promoters. In vivo studies on the experimental model are ongoing, and results will be presented.

No significant financial relationships to disclose.
Targeted therapy for osteosarcoma using 17AAG and rapamycin: Identification of new targets using Gene Microarray profiling and MiRNA
Authors: Gazitt Y. , Moncada K. , Thomas C. , Takimoto C. , Tolcher A.
Published: March 2020

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Background: Sarcomas constitute a group of diverse diseases with many subtypes and varying prognoses. Hence, new approaches for therapy involving targeting of specific survival pathways are needed. Methods: We used 17-AAG to block HSP90 and rapamycin (Ra) to block mTOR, in 2 osteosarcoma cell lines, HOS and KHOS/NP. Cells were treated for 24 and 48 hours with 200 nM of 17- AAG or 5 micromolar of rapamycin. Gene profiling (GP) was performed using h-U133 2 Plus Array (Affymetrix). Results were pre-filtered using the GCOS pair-wise comparisons. ANOVA was filtered for >2-fold change between two conditions. We compared control HOS or KHOS vs. 24 H or 48 H treatment with Ra, or 17AAG. Gene identity, ontology and z-scores were derived. For micro RNAs (miRs), we used a miRNA chip (LC Sciences) and data were normalized, adjusted and clustered using t-test (p<0.01). Results and Conclusions: 17-AAG is a good inducer of apoptosis in HOS and KHOS cells (IC50 of ∼50nM) and blocked cells at mitosis. Ra is a weak inducer of apoptosis (IC20 at 5 micromolar) and blocked cells at G1. GP of 17AAG treated HOS/KHOS cells revealed >1,000 and 1,600 gene changes at 24 H and 48 H, respectively, with ∼700 genes affected at 24 and 48H. Ra affected ∼400 and ∼350 genes at 24 and 48H, respectively, with 20 genes affected at 24 and 48H. Of the genes upregulated by 17AAG, >450 were in cell and protein metabolism; >50 genes were in chromatin modifications and >40 in regulation of cell cycle. With 17AAG, changes were found in histones and deacetylases; cytoskeleton proteins; extracellular matrix proteins and MMPs; HSPs; MAP kinase, mTOR, IGF and VEGF pathways; cell cycle and apoptosis. Many targets were validated by by immunoblotting. MiRs were profiled following 24H treatment with 17AAG or Ra. 17AAG upregulated miR122a;145;143 and 663 (4–16 fold) and miR 542–5P was decreased (∼12 fold). Ra upregulated miR 498;145;143;452;663; 602 and miR224 (4–32 fold) and downregulated (4–32 fold) miR1;542–5P and miR335. Taken together, miR changes affect the expression of transcripts related to angiogenesis; cell cycle; cell signaling and apoptosis and are common to the changes observed by GP. In summary, new therapeutic targets were identified by the combination of GP and miRNA following 17AAG or rapamycin.

No significant financial relationships to disclose.
STEAP1: A potential marker of metastatic disease for Ewing family tumors
Authors: Cheung I. Y. , Feng Y. , Danis K. , Shukla N. , Meyers P. , Ladanyi M. , Cheung N.
Published: March 2020

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Background: Ewing Family Tumors (EFTs) metastasize by hematogeneous spread. Because of its dismal cure rate, targeting subclinical disease is particularly relevant. Genome-wide gene expression array analysis can uncover novel genes differentially expressed in tumors over normal marrow/blood, which may have clinical potentials as markers of minimal residual disease (MRD). Methods: Gene expression array data were obtained on 28 EFT tumors and 10 EFT cell lines using the Affymetrix U133 gene chip. Ten genes with high tumor to blood ratios were identified. Quantitative RT-PCR was performed to study (1) the dynamic range of detection of rare tumor cells (frequency of 10-7 to 10-4), (2) the gene expression in normal blood and marrow samples, (3) the gene expression among EFT tumors, and (4) the prognostic impact of marker positivity in EFT patients with histologically negative marrows. Results: Of the 10 genes, 5 genes, namely six transmembrane epithelial antigen of the prostate 1 [STEAP1], cyclin D1 [CCND1], plakophilin 1 [PKP1], transmembrane protein 47 [TMEM47], and NKX2.2 transcription factor [NKX2.2] were chosen for further analyses. This was based on their steep linear dynamic range in tumor cells seeded in normal mononuclear cells and consistently high expression among EFT tumor samples. Among 53 EFT patients with histologically negative marrow samples, a marker was classified as positive if the gene transcript level was above the upper limit of normal, defined as mean + 2SD of 26 normal blood and marrow samples. Of the 5 markers tested, STEAP1 positivity in marrow samples was highly correlated with survival with new metastasis in Kaplan Meier analysis (p=0.001). Conclusions: This gene expression array-based approach identified STEAP1 as a promising surrogate MRD marker of metastatic EFT with potential prognostic importance. Its clinical utility will need to be further tested in large patient cohorts.

No significant financial relationships to disclose.