bluuebunny/crossref_metadata_2025 huggingface dataset:
A core outcome set for lower limb orthopaedic surgery for children with cerebral palsy: An international multi‐stakeholder consensus study
Authors: Almoajil Hajar , Hopewell Sally , Dawes Helen , Toye Francine , Theologis Tim
Published: July 2022

Aim

To develop a core set of outcome domains to be measured in clinical studies on lower limb orthopaedic surgery for ambulant children with cerebral palsy (CP) that represents the priorities of an international multi‐stakeholder group (children, parent/carers, and health professionals).

Method

Potential outcome domains were identified through literature review and qualitative interviews with key stakeholders. These were scored in an international two‐round Delphi survey, using a 9‐point Likert scale. A final consensus meeting with key stakeholders agreed on the most important outcome domains and refined the core outcome set (COS).

Results

One hundred and sixty‐one health professionals and 36 individuals with CP and their parents/carers rated 21 of 41 outcomes as important in the Delphi survey. The final consensus group agreed 19 outcomes within eight domains to be included in the final COS: pain and fatigue, lower limb structure, motor function, mobility (daily life activities), gait‐related outcomes, physical activity, independence, and quality of life.

Interpretation

A COS for lower limb orthopaedic surgery for children with CP was developed. Incorporating this in the design of future clinical studies will provide a more holistic assessment of the impact of treatment while allowing meaningful comparisons and future synthesis of results from primary studies.

What this paper adds

Eight core outcome domains were identified as important to measure in future clinical research.

Key stakeholders perceived pain, balance and fall, and independence as very important outcomes.

Six contextual factors were identified as essential in surgical decision‐making.
Nutrition and food literacy among young Canadian adults living with type 1 diabetes
Authors: Itzkovitz Alexandra , Maggio Vanessa , Roy‐Fleming Amélie , Legault Laurent , Brazeau Anne‐Sophie
Published: July 2022

Aim

Nutrition and food literacy encompasses knowledge, skills and confidence to prepare healthy meals. This project aimed to assess and compare the proportion of young Canadian adults (18–29 years old) living with type 1 diabetes and without diabetes (controls) who demonstrated adequate nutritional health literacy.

Methods

This cross‐sectional study involved participants completing an online survey that included questions on socio‐economic status, nutrition knowledge, confidence and skills in meal preparation and the Short Food Literacy Questionnaire (SFLQ). Proportion of participants with adequate SFLQ score (i.e. ≥34/52) was compared between the groups (two‐sample t‐test).

Results

Among the 236 people living with type 1 diabetes and 191 controls (81.5% women), mean age was 24 ± 3 years for people living with type 1 diabetes and 22 ± 3 years for controls (p < 0.001). More people living with type 1 diabetes reported adequate SFLQ score (people living with type 1 diabetes 88.0% vs. Controls 68.0%; p < 0.001). Similarly, majority of people living with type 1 diabetes prepared their own meals compared to the controls (74.5% vs. 47.6%; p < 0.001). Enhanced SFLQ score was associated with higher cooking skills (p = 0.02) and confidence (p < 0.01) in preparing healthy meals.

Conclusions

Living with type 1 diabetes was associated with greater SFLQ scores among young Canadian adults. Having the independence, the confidence and skills in meal preparation were contributing factors.
Sources of occupational stress amongst dentistry academics
Authors: Teixeira Hugo , Lalloo Ratilal , Evans Jane L. , Fuss Janet , Pachana Nancy A. , Burton Nicola W.
Published: July 2022

Background

Dentistry professionals may experience significantly higher occupational stress than other health professionals and dentistry academics may have specific work content and context sources of stress.

Aims

The aim of this study is to identify common sources of occupational stress, and how these are associated with wellbeing, in dentistry academics.

Materials & Methods

A cross‐sectional online survey with staff in Dentistry departments in Australia and New Zealand. Assessment included 23 items from five general domains of occupational stress from the NIOSH—Generic Job Stress Questionnaire, a 23‐item list of sources of stress and the 22‐item Psychological General Well‐Being Index. Analyses used descriptive statistics and multiple linear regression.

Results

A total of 107 respondents (average age 50 ± 11.7 years, 56.8% men) completed the survey. Leading sources of occupational stress were job future, time pressure at work, work overload, and administration demands. A multiple linear regression model significantly predicted wellbeing, F(8,77) = 13.141, p = .000, adj.R2 = .53, but there were no significant associations for any of the specific sources of stress.

Conclusion

The combination of time pressure, workload and responsibility, job dissatisfaction, low social support, and uncertain job future was inversely associated with wellbeing amongst these dentistry academics. Future studies should consider the development and evaluation of interventions to address these concerns.
Investigating periodontal diagnosis and treatment at one dental school
Authors: DeStaffany Amy M. , Gurenlian Jo Ann R. , Bono Leciel K.
Published: July 2022

Introduction

The purpose of this study was to evaluate whether patients at one dental education institution received appropriate care based on their periodontal diagnosis in a timely manner.

Materials and Methods

This study used a retrospective design to evaluate data from July 2018 to February 2020. The patient records were screened to determine whether patients received a periodontal diagnosis, whether the diagnosis followed the 2018 AAP Classification Guidelines, and if appropriate preventive and therapeutic procedures were completed in a timely manner. In addition, the type or year of dental student providing the procedure was also recorded. Data were analysed using descriptive statistics and a Chi‐squared test.

Results

A total of 612 charts were generated for review and 157 met the inclusion criteria and were evaluated. Results revealed that more than half (56.7%) of the patient records did not demonstrate a periodontal diagnosis and another 10.8% did not follow current AAP Classification Guidelines. Most patients (n = 125, 79.6%) had a comprehensive periodontal assessment performed, while 32 (20.4%) had no comprehensive periodontal charting information recorded. Further, of the 157 records reviewed, 96 (61.1%) had no periodontal treatment specified. A statistically significant difference with a modest association was found between type of dental student and year based on periodontal diagnosis (Fisher's exact test value = 20.72, p = .001, Cramer's V = 0.25).

Conclusion

Documentation of key clinical information, diagnostic conclusions, and treatment rendered requires curriculum review. Further studies are warranted to determine whether similar findings exist among other dental school education programs.
Impact of public health and higher education policies on the profile of final‐year Brazilian dental students: Challenges and future developments
Authors: Bitencourt Fernando Valentim , Olsson Thaís Ostroski , Lamers Juliana Maciel de Souza , Leite Fabio Renato Manzolli , Nascimento Gustavo Giacomelli , Toassi Ramona Fernanda Ceriotti
Published: July 2022

Introduction

Brazil has experienced transformations in higher education and health services, including launching more inclusive public policies focused on these two areas.

Objective

To evaluate the profile of final‐year dental students from a Brazilian public university from 2010 to 2019, accompanied by changes in public health and higher education policies.

Methods

A prospective observacional study was carried out with final‐year dental students. A self‐applicable semi‐structured questionnaire was applied.

Result

Six‐hundred and seventy‐seven students participated, of which 71.5% were women, 72.9% aged between 21 and 25 years, 96.2% single, and 96.4% were without children. Over ten years, it was possible to identify trends in the profile explained by implementing public policies toward more inclusive access to Brazilian higher education by socioeconomically disadvantaged individuals. Students who completed the course between 2018 and 2019 did not have the state capital city (the wealthiest area) as their origin city and had lower parental education and income levels than dental students graduating between 2010 and 2011. Moreover, working as a primary care dentist in the Brazilian National Health System was considered a professional possibility by 61.4% of the students, and has gained prominence significantly over time, ranging from 21.1% in 2010 to 72.9% in 2019 (p < .05).

Conclusion

Over the study period, concomitantly to advances in public health and higher education policies in Brazil, more diverse access to public dental education was observed, allowing students from low socioeconomic positions to take the education. Changes have also impacted the students' perspectives regarding the need for postgraduate training and a career in public health dentistry. However, these trends need to be consolidated, and public policies continued and strengthened.
Doctoral education from its medieval foundations to today's globalisation and standardisation
Authors: Ruano‐Borbalan Jean‐Claude
Published: July 2022

The doctorate, as a ritualised form of evaluation, has existed for more than eight centuries—initially linked to Western medieval forms of knowledge production framed by religion, and to the professional training of lawyers and doctors. It nearly disappeared at the end of the modern era. Recast as a requirement for academic professions, it proceeded to play an important role in the production of new knowledge and became a key to scientific and research development in the 19th and 20th centuries. Since less than half a century ago, doctoral education has gone through tremendous changes due to both the globalisation and standardisation of higher education and public policies designed to encourage knowledge transfers from academia to society at large. Doctoral education has become a crucial part of knowledge production and the transfer of knowledge into the broader economy, but it faces huge obstacles that can only be understood by taking into account the history of its transformations, the anthropological role it played and still plays, and also the controversial effect of efforts to standardise knowledge production, which goes against the non‐standard nature of human knowledge creation.
Machine learning validation through decision tree analysis of the Epidemiology‐Based Mortality Score in Status Epilepticus
Authors: Brigo Francesco , Turcato Gianni , Lattanzi Simona , Orlandi Niccolò , Turchi Giulia , Zaboli Arian , Giovannini Giada , Meletti Stefano
Published: July 2022

Objective

This study was undertaken to validate the accuracy of the Epidemiology‐Based Mortality Score in Status Epilepticus (EMSE) in predicting the risk of death at 30 days in a large cohort of patients with status epilepticus (SE) using a machine learning system.

Methods

We included consecutive patients with SE admitted from 2013 to 2021 at Modena Academic Hospital. A decision tree analysis was performed using the 30‐day mortality as a dependent variable and the EMSE predictors as input variables. We evaluated the accuracy of EMSE in predicting 30‐day mortality using the area under the receiver operating characteristic curve (AUC ROC), with 95% confidence interval (CI). We performed a subgroup analysis on nonhypoxic SE.

Results

A total of 698 patients with SE were included, with a 30‐day mortality of 28.9% (202/698). The mean EMSE value in the entire population was 57.1 (SD = 36.3); it was lower in surviving compared to deceased patients (47.1, SD = 31.7 vs. 81.9, SD = 34.8; p < .001). The EMSE was accurate in predicting 30‐day mortality, with an AUC ROC of .782 (95% CI = .747–.816). Etiology was the most relevant predictor, followed by age, electroencephalogram (EEG), and EMSE comorbidity group B. The decision tree analysis using EMSE variables correctly predicted the risk of mortality in 77.9% of cases; the prediction was accurate in 85.7% of surviving and in 58.9% of deceased patients within 30 days after SE. In nonhypoxic SE, the most relevant predictor was age, followed by EEG, and EMSE comorbidity group B; the prediction was correct in 78.9% of all cases (89.6% in survivors and 46.1% in nonsurvivors).

Significance

This validation study using a machine learning analysis shows that the EMSE is a valuable prognostic tool, and appears particularly accurate and effective in identifying patients with 30‐day survival, whereas its performance in predicting 30‐day mortality is lower and needs to be further improved.
Novel distance and entropy definitions for linear Diophantine fuzzy sets and an extension of TOPSIS (LDF‐TOPSIS)
Authors: Gül Sait , Aydoğdu Ali
Published: July 2022

The literature of multiple attribute decision making (MADM) is fruitful since there are various and successful applications of different fuzzy set extensions such as intuitionistic, Pythagorean and q‐Rung orthopair fuzzy sets (IFS, PFS and q‐ROFS). Besides their powerful aspects, some definitional limitations are known. In order to eradicate these boundaries regarding the definitions of membership and non‐membership degrees, linear Diophantine fuzzy set (LDFS) concept has been recently emerged. By considering two parameters, LDFS extends the representation area of the previous fuzzy set definitions and provides more extensive human judgement coverage field. In this study, the first distance and entropy measures in the literature have been developed for LDFSs. Their axiomatic definitions are given, and the proofs are shown. Also, thanks to our extensive literature review, we became aware that there is no MADM extension dedicatedly proposed for LDFS. So, the first MADM method extension for LDFS environment has also been developed in this study. A very well‐known MADM approach, TOPSIS, has been extended into LDFS environment for the first time in the literature. The applicability is shown in a healthcare management decision problem and the validity is checked and approved by comparing the alternative rankings LDF‐TOPSIS and the aggregation operators that were obtained from the literature produced.
Role of the Paf1 complex in the maintenance of stem cell pluripotency and development
Authors: Park Jiyeon , Park Shinae , Lee Jung‐Shin
Published: July 2022

Cell identity is determined by the transcriptional regulation of a cell‐type‐specific gene group. The Paf1 complex (Paf1C), an RNA polymerase II‐associating factor, is an important transcriptional regulator that not only participates in transcription elongation and termination but also affects transcription‐coupled histone modifications and chromatin organisation. Recent studies have shown that Paf1C is involved in the expression of genes required for self‐renewal and pluripotency in stem cells and tumorigenesis. In this review, we focused on the role of Paf1C as a critical transcriptional regulator in cell fate decisions. Paf1C affects the pluripotency of stem cells by regulating the expression of core transcription factors such as Oct4 and Nanog. In addition, Paf1C directly binds to the promoters or distant elements of target genes, thereby maintaining the pluripotency in embryonic stem cells derived from an early stage of the mammalian embryo. Paf1C is upregulated in cancer stem cells, as compared with that in cancer cells, suggesting that Paf1C may be a target for cancer therapy. Interestingly, Paf1C is involved in multiple developmental stages in Drosophila, zebrafish, mice and even humans, thereby displaying a trend for the correlation between Paf1C and cell fate. Thus, we propose that Paf1C is a critical contributor to cell differentiation, cell specification and its characteristics and could be employed as a therapeutic target in developmental diseases.
Limited spatial rescue potential for coral reefs lost to future climate warming
Authors: Greiner Ariel , Andrello Marco , Darling Emily , Krkošek Martin , Fortin Marie‐Josée
Published: July 2022

Aim

The aim was to determine reef connectivity and future coral cover levels under global scenarios of coral bleaching loss and potential recovery.

Location

Global coral reefs.

Time period

Present‐day to 2100.

Major taxa studied

Scleractinian coral.

Methods

We used a global coral larval dispersal model that describes population connectivity among reefs at a resolution of ⅙° × ⅙° (c. 18 km × 18 km) cells. To simulate different patterns of bleaching events, we ran three scenarios at different levels of coral reef habitat loss followed by a reseeding of coral larvae from surviving reefs to simulate recovery.

Results

We found a total of 604 distinct reef networks, but more than half of the world's reef cells are contained in six large coral reef networks (294–5,494 cells), whereas the rest form smaller networks. In the bleaching scenario where previously identified predicted climate refugia were maintained, initial connectivity was largely preserved even when 71% of global coral reef habitat was lost, but the relict reef cells were unable to reseed even 50% of former coral reef habitat because many of the relict reefs are in the same networks as each other. In scenarios where refugia were lost first or with random loss, less of the initial connectivity was maintained, but more widespread reseeding was possible because more reef cells within smaller networks were maintained.

Main conclusions

Our findings highlight the importance of maintaining functional coral reef habitat outside of predicted climate refugia to sustain connectivity globally, and suggest an important role for “stepping stone” reefs between the climate refugia. Without attention to these issues of habitat loss and connectivity, much of global coral reef habitat might not be reseeded without human intervention.
Dataset for pathology reporting of ductal carcinoma in situ, variants of lobular carcinoma in situ and low‐grade lesions: recommendations from the International Collaboration on Cancer Reporting (ICCR)
Authors: Fox Stephen B , Webster Fleur , Chen Chih‐Jung , Chua Boon , Collins Laura , Foschini Maaria‐Pia , Bruce Mann G , Millar Ewan , Pinder Sarah E , Rakha Emad , Shaaban Abeer , Tan Benjamin Y , Tse Gary , Watson Peter , Tan Puy Hoon
Published: July 2022

Aims

To describe a new international dataset for pathology reporting of ductal carcinoma in situ (DCIS), variants of lobular carcinoma in situ (LCIS) and low‐grade lesions (encapsulated papillary carcinoma, solid papillary carcinoma in situ, Paget's disease) produced by the International Collaboration on Cancer Reporting (ICCR).

Methods and results

The ICCR, a global alliance of pathology bodies, uses a rigorous and efficient process for the development of evidence‐based, structured datasets for pathology reporting of common cancers. Their aim is to support quality pathology reporting and engender understanding between the breast surgeon, pathologist, and oncologist for optimal and uniform patient management globally. Here we describe the dataset for DCIS, some variants of LCIS (namely the pleomorphic and the florid variants), and low‐grade lesions by a multidisciplinary panel of internationally recognized experts. The agreed dataset comprises 12 core (required) and five noncore (recommended) elements suitable for both developed and low‐income jurisdictions, derived from a review of current evidence. Areas of contention were addressed using a pragmatic approach in the absence of evidence. Use of all core elements is the minimum reporting standard for any individual case. Commentary is provided, explaining each element's clinical relevance, definitions to be applied where appropriate for the agreed list of value options and the rationale for considering the element as core or noncore.

Conclusion

This first internationally agreed dataset for DCIS, variants of LCIS, and low‐grade lesions reporting will enable their standardization of pathology reporting and enhance clinicopathological communication leading to improved patient outcomes. Widespread adoption will also facilitate international comparisons, multinational clinical trials, and help to improve the management of breast disease globally.
Microstructure design for bone void filler
Authors: Tuan Wei‐Hsing , Chen Ying‐Cen , Hsu Pei‐Yi , Chang Hao‐Yu , Yeh Li‐Yun , Chen Chih‐Yi , Lai Po‐Liang
Published: July 2022

Bone void fillers are used to fill in the empty space within bone defect. The use of fillers is aiming to reduce the time required for healing the defect. The filler should show the resistance to washout and the ability of adsorption within human body. In the present study, a microstructure design approach is employed to evaluate the performance of bone void filler. The microstructure features, such as the contacts between particles, the size of pores, and their interconnectivity, are considered. The pellets composing of these microstructure features are prepared for in vitro and in vivo evaluations. The type of contacts between resorbable particles plays a key role on washout resistance. The interconnectivity of pores affects significantly on the absorption rate in vivo. Through the control of these microstructure characteristics, the resorbable behavior of bone void filler can be tailored.
Practices, challenges, and impacts of teaching and learning French as a second language in Ontario secondary schools: Focus on tech‐oriented students
Authors: Takam Alain Flaubert , Gillis Steven A.
Published: July 2022

This paper examines the practices, challenges, and future potential of the French as a second language (FSL) program in the Canadian province of Ontario's secondary schools, with a particular focus on those interested in technological and business programs. Questionnaires were filled out by students, parents, and teachers in six secondary schools. The responses offered insights into the current situation in Ontario's FSL, as well as into the possible utility of, and desire for, the development of FSL courses aimed at students learning in technological and business fields. The findings and their potential implications for FSL in Ontario will be presented.
Mendelian randomization analysis identified causal Association of Childhood Obesity with adult major depressive disorder
Authors: Yan Shan‐Shan , Xu Qian , Han Bai‐Xue , Ni Jing‐Jing , Wei Xin‐Tong , Feng Gui‐Juan , Zhang Hong , Zhang You‐Jie , Zhang Lei , Yu Wen‐Yuan , Pei Yu‐Fang
Published: July 2022

Background

Childhood obesity is associated with adult major depressive disorder (MDD), but their causality is not clear.

Methods

We performed a two‐sample Mendelian randomization (MR) analysis to explore the causality of childhood body mass index (BMI) and childhood obesity on MDD, followed by a multivariable MR (MVMR) analysis to investigate the potential role of adult BMI in mediating such effect. We accessed genome‐wide association summary statistics of childhood BMI, childhood obesity, adult BMI and adult MDD from the Early Growth Genetics consortium (nBMI = 47 541, nobesity = 24 160), the Genetic Investigation of Anthropometric Traits consortium (nadult_BMI = ∼700 000) and the Psychiatric Genomics consortium (nMDD = 500 199), respectively. The MR‐PRESSO test was performed to remove SNPs with potential pleiotropic effect. The MR analysis was performed by inverse‐variance weighted test. Further sensitivity analyses, including the MR‐Egger intercept test and leave‐one‐out analysis, were performed to evaluate the reliability of the results.

Results

Our study found that childhood obesity might increase the odds of developing MDD in adults (OR = 1.03, 95% CI: 1.01–1.06, p = 2.6 × 10−3). Children with higher BMI were more likely to develop MDD in adulthood, with an OR of 1.12 per standard deviation score (SDS) increase in BMI (95% CI: 1.07–1.17, p = 4.4 × 10−7). Sensitivity analyses verified the reliability of the causality between childhood BMI/obesity and MDD. Further MVMR results revealed that the impact of childhood BMI on MDD risk was predominantly mediated by adult BMI.

Conclusion

Our findings provided evidence of a causal relationship between childhood BMI/obesity and adult MDD, thus providing new insights into the prevention of MDD.
Embracing brain and behaviour: Designing programs of complementary neurophysiological and behavioural studies
Authors: Kirwan C. Brock , Vance Anthony , Jenkins Jeffrey L. , Anderson Bonnie Brinton
Published: July 2022

NeuroIS—the methods and knowledge of neuroscience applied to the information systems (IS) domain—has become an established research field within the IS discipline. A key advantage of NeuroIS is its ability to provide insights into human cognition beyond those obtained using behavioural techniques alone. Nevertheless, in neuroscience, there is renewed interest in examining behaviour together with neurophysiological methods to better inform our understanding of neural processes. In this research opinion article, we argue that in the field of NeuroIS, there is an opportunity for hybrid programs of study that combine neurophysiological and behavioural methods in a complementary manner. We outline four strategies for designing complementary neurophysiological and behavioural experiments in a research program: (1) observe the relationship between neural processes and behavioural change; (2) combine neurophysiological and behavioural methods to enhance internal, external, and ecological validity; (3) extend, rather than replicate, experiments based on theory; and (4) use neurophysiological and behavioural experiments together to evaluate IT artefact design. By applying these strategies, researchers can more effectively design programs using complementary neurophysiological and behavioural methods, which, in turn, can help to provide richer insights into the phenomena under study as well as accelerate the advancement of IS knowledge.
Insight into the spectroscopic behavior of Dy3+‐substituted and Li+ charge–compensated molybdate phosphors
Authors: Nayak Priyanka , Nanda Sushri Sangita , Gupta Santosh K. , Sudarshan K. , Dash S.
Published: July 2022

Efficient utilization of phosphors in solid‐state lighting and phosphor‐converted light‐emitting diodes demands high emission intensity, narrow size distribution, and tunable luminescence. Here in this work, un‐doped and Li+ co‐doped CaMoO4:Dy3+ nanoparticles are synthesized via modified reflux method in ethylene glycol medium at 130°C. The host shows broad dual band emission centered in blue and green regions endowed by charge transfer and defects. Upon Dy3+ doping, the host‐to‐dopant energy transfer causes the CaMoO4:Dy3+ nanoparticles to exhibit characteristic emission lines from Dy3+. The Commission Internationale de L'Eclairage (CIE 1931) chromaticity coordinates for the prepared phosphors are estimated and found to lie in the nearly white region of color space. The light output as well as excited state lifetimes are successfully improved by lithium co‐doping by virtue of reduced charge compensating defects and sensitization via oxygen‐related defects. The positron annihilation studies show that all the lithium concentration co‐doped for charge compensation is merely helping in the removal of cations. This work shows the role of lithium co‐doping and its optimum concentration in improving the optical properties of phosphors.
Ultrafast charge carrier dynamics and photoluminescence of CsPbBr3−xIx quantum dots in boro‐germanate glasses
Authors: Liu Yingrui , Wang Jing , Han Jianjun , Liu Chao , Ruan Jian
Published: July 2022

CsPbBr3−xIx quantum dots were precipitated in boro‐germanate glasses with various Br/I ratios by the melt‐quenching method. When the heat treatment temperature (TH) increased from 480 to 550°C, more I− entered the perovskite crystal lattice, resulting in lattice expansion, the X‐ray diffraction peaks of CsPbBr1.5I1.5 glasses shifted to small angles. The high concentration of I− entered into the perovskite crystal lattice, decreasing the bandgap. The photoluminescence (PL) peaks of CsPbBr1.5I1.5 glasses redshifted from 577 to 620 nm, the PL peaks of CsPbBrI2 glasses shifted from 614 to 636 nm, and the PL peaks of CsPbBr0.5I2.5 glasses shifted from 650 to 672 nm. As the ratio of Br/I reduced from 2.5/0.5 to 0.5/2.5, the PL lifetime of CsPbBr3−xIx glass increased from 2.08 to 74.5 ns. With the delay time from 1 to 1500 ps, the carriers gradually accumulated in the band‐edge state, and the bleaching signal intensity increased. Then more and more carriers returned to the ground state through radiative or nonradiative recombination, the bleaching signal intensity decreased.
Purification and characterization of a robust thermostable protease isolated from Bacillus subtilis strain HR02 as an extremozyme
Authors: Homaei Ahmad , Izadpanah Qeshmi Fatemeh
Published: July 2022

Aim

Since the hot water of Genow, a village in Isin rural district in the central district of Bandar Abbas, Hormozgan Province, Iran, has a rich source of thermophilic bacteria, the current study aimed to find a new thermophilic protease enzyme with suitable properties to be used in different industries.

Methods and results

Water and sediment samples were collected from the hot water of Genow, and finally, 20 colonies were isolated. Among these isolated colonies, two bacterial strains grew on the skim milk agar medium, and a clear halo was formed around the colony, which was accurately identified by 16S rRNA gene sequence analysis. The comparison of 16S rRNA gene sequence analyses of isolated strains HR01 and HR02 with registered sequences of 16S rRNA genes in NCBI showed that the two isolates had the most similarity to Bacillus sonorensis and Bacillus subtilis, respectively. Among the two bacterial strains, the highest enzymatic activity was observed in B. subtilis strain HR02, from which the protease purification process was performed. A putative native B. subtilis strain HR02 protease (BSHR02PR) was purified by the UNO Q-6 ionic exchange chromatography method. Biochemical analyses revealed a monomeric enzyme, BsHR02Pro, with a molecular weight of 25 kDa, showing the maximum activity at 70°C and pH 8.0. Moreover, the purified enzyme was stable up to 80 °C and in a pH range of 6.0–12.0. The steady-state kinetic analysis for colloidal casein showed that the Km, Vmax and kcat values of the purified enzyme were 25.7 μM, 93.2 μM min−1 and 2.18 s−1, respectively.

Conclusion

The hot water of Genow is a rich source of protease-producing bacteria. Sediments are a better source for the isolation of these types of bacteria than spring water. Overall, our results demonstrated a potential bacterial enzyme BsHR02Pro as a suitable catalyst to be used in the various industries.
Comparative metabolomics and transcriptomics analyses provide insights into the high-yield mechanism of phenazines biosynthesis in Pseudomonas chlororaphis GP72
Authors: Li Song , Yue Sheng-Jie , Huang Peng , Feng Tong-Tong , Zhang Hong-Yan , Yao Rui-Lian , Wang Wei , Zhang Xue-Hong , Hu Hong-Bo
Published: July 2022

Aims

Phenazines, such as phenazine-1-carboxylic acid (PCA), phenazine-1-carboxamide (PCN), 2-hydroxyphenazine-1-carboxylic acid (2-OH-PCA), 2-hydroxyphenazine (2-OH-PHZ), are a class of secondary metabolites secreted by plant-beneficial Pseudomonas. Ps. chlororaphis GP72 utilizes glycerol to synthesize PCA, 2-OH-PCA and 2-OH-PHZ, exhibiting broad-spectrum antifungal activity. Previous studies showed that the addition of dithiothreitol (DTT) could increase the phenazines production in Ps. chlororaphis GP72AN. However, the mechanism of high yield of phenazine by adding DTT is still unclear.

Methods and Results

In this study, untargeted and targeted metabolomic analysis were adopted to determine the content of metabolites. The results showed that the addition of DTT to GP72AN affected the content of metabolites of central carbon metabolism, shikimate pathway and phenazine competitive pathway. Transcriptome analysis was conducted to investigate the changed cellular process, and the result indicated that the addition of DTT affected the expression of genes involved in phenazine biosynthetic cluster and genes involved in phenazine competitive pathway, driving more carbon flux into phenazine biosynthetic pathway. Furthermore, genes involved in antioxidative stress, phosphate transport system and mexGHI-opmD efflux pump were also affected by adding DTT.

Conclusion

This study demonstrated that the addition of DTT altered the expression of genes related to phenazine biosynthesis, resulting in the change of metabolites involved in central carbon metabolism, shikimate pathway and phenazine competitive pathway.

Significance and Impact of the Study

This work expands the understanding of high yield of phenazine by the addition of DTT and provides several targets for increasing phenazine production.
The antibacterial potency and antibacterial mechanism of a commercially available surface-anchoring quaternary ammonium salt (SAQAS)-based biocide in vitro
Authors: Saseendran Nair Shilpa , Anand Vikash , De Silva Karnika , Wiles Siouxsie , Swift Simon
Published: July 2022

Aims

To determine the antimicrobial potency of a surface-anchored quaternary ammonium salt (SAQAS)-based biocide during in vitro wet and dry fomite assays and to determine the mechanism of killing bacteria on the surface.

Methods and Results

Wet and dry fomite assays were established in vitro for a commercially available biocide (SAQAS-A) applied to glass and low-density polyethylene (LDPE) surfaces. Both wet and dry fomite tests showed the active killing of Gram-positive and Gram-negative bacteria but not endospores. Assays measuring membrane permeability (ATP and DNA release), bacterial membrane potential and bacterial ROS production were correlated with the time-to-kill profiles to show SAQAS-A activity in suspension and applied to a surface.

Conclusions

SAQAS-A is an effective biocide against model strains of vegetative bacteria. The killing mechanism for SAQAS-A observed minimal membrane depolarization, a surge in ROS production and assessment of membrane permeability supported the puncture of cells in both suspension and surface attachment, leading to cell death.

Significance and Impact of the study

SAQAS represents effective surface biocides against single challenges with bacteria through a mechanical killing ability that supports real-world application if their durability can be demonstrated to maintain residual activity.
Complete-genome sequence and in vitro probiotic characteristics analysis of Bifidobacterium pseudolongum YY-26
Authors: Yu Ying , Ren Xinmiao , Cao Linyuan , Liang Qingping , Xiao Mengshi , Cheng Jiaying , Nan Shihao , Zhu Changliang , Kong Qing , Fu Xiaodan , Mou Haijin
Published: July 2022

Aims

The aim was to isolate a neotype bifidobacteria strain and evaluate its in vitro probiotic potential.

Methods and Results

Bifidobacterium pseudolongum YY-26 (CGMCC 24310) was isolated from faeces of mice treated with low-molecular-weight hydrolyzed guar gum (GMPS) and identified based on 16S rRNA sequence and genome sequence. Whole-genome sequencing obtained using PacBio's single-molecular and Illumina's paired-end sequencing technology. A genome of 2.1 Mb in length, with 1877 predicted protein-coding sequences was obtained. Carbohydrate-Activity enZyme analysis revealed that YY-26 encodes 66 enzymes related to carbohydrate metabolism. Whole genome sequence analysis revealed the typical probiotic characteristics of YY-26, including safety in genetic level and ability to produce beneficial metabolites and extracellular polysaccharides. Ability of extensive carbon source utilization and short-chain fatty acid production was observed with single YY-26 cultivation. Considerable acetic acids and lactic acids were determined in GMPS utilization. YY-26 showed tolerance to simulated gastrointestinal tract and displayed appreciable antioxidant activity of free radical scavenging.

Conclusions

B. pseudolongum YY-26 was identified with numerous probiotic-associated genes and its probiotic characteristics were verified in vitro.

Significance and Impact of Study

This study supplemented with limited publicly information regarding the genomes of B. pseudolongum strains and revealed the probiotic potential of YY-26.
High-throughput RNA sequencing and genetic structure studies of turnip mosaic virus infecting black and yellow mustard revealing emergence of world-B3 pathotype in India
Authors: Singhal Pankhuri , Baranwal Virendra Kumar , Prajapati Malyaj R. , Singh Jitender
Published: July 2022

The RNA viruses are marked by high genetic diversity, which allows them to quickly adapt to new and resistant hosts. The pathogenic turnip mosaic virus (TuMV) infects Brassicaceae plant species all over the world.

Aim

To study the evolution and host expansion of a TuMV for the first time in India using molecular population genetic framework.

Materials and Results

Here, we decipher the complete genome sequences of two TuMV world-B3 strains infecting yellow and black mustard in India through high-throughput RNA sequencing subjecting ribosomal RNA depleted mRNA isolated from leaves exhibiting puckering and mosaic symptoms with 100% incidence and high severity in the experimental field. The viral genomes of the two isolates were 9817 and 9829 nucleotides long. They featured two open reading frames (ORFs), one of which encoded a polyprotein comprised of 3164 amino acids and the other of which encoded a PIPO protein of 62 amino acids.

Conclusions

The two TuMV strains from New Delhi region shared identity with the world-B pathotype and subpathotype world B3 showcasing its emergence first time in South Asia. In contrast, other isolates reported previously from South Asia were all Asian-BR pathotypes.

Significance and Impact of the Study

According to our knowledge, this is the first instance of TuMV association with black mustard naturally. Their geographical prevalence justifies a lower degree of genetic differentiation and higher rate of gene flow calculated between the world-B and Asian-BR pathotypes. This study provides insights on population structuring, expansions and evolution, level of genetic heterogeneity and variability of worldwide prevalent isolates of TuMV which will further aid in understanding virus epidemiology and help prevent losses.
AaCaM is required for infection structure differentiation and secondary metabolites in pear fungal pathogen Alternaria alternata
Authors: Jiang Qianqian , Mao Renyan , Li Yongcai , Bi Yang , Liu Yongxiang , Zhang Miao , Li Rong , Yang Yangyang , Prusky Dov B.
Published: July 2022

Aims

Calmodulin (CaM), acts as a kind of multifunctional Ca2+ sensing protein, which is ubiquitous in fungi, is highly conserved across eukaryotes and is involved in the regulation of a range of physiological processes, including morphogenesis, reproduction and secondary metabolites biosynthesis. Our aim was to understand the characteristics and functions of AaCaM in Alternaria alternata, the causal agent of pear black spot.

Methods and results

A 450 bp cDNA sequence of AaCaM gene of A. alternata was cloned by the PCR homology method. Sequence analysis showed that this protein encoded by AaCaM was a stable hydrophilic protein and had a high similarity to Neurospora crassa (CAA50271.1) and other fungi. RT-qPCR analysis determined that AaCaM was differentially upregulated during infection structural differentiation of A. alternata both on hydrophobic and pear wax extract-coated surface, with a 3.37-fold upregulation during the hydrophobic induced appressorium formation period (6 h) and a 1.46-fold upregulation during the infection hyphae formation period (8 h) following pear wax induction. Pharmaceutical analysis showed that the CaM-specific inhibitor, trifluoperazine (TFP), inhibited spore germination and appressorium formation, and affected toxins and melanin biosynthesis in A. alternata.

Conclusions

AaCaM plays an important role in regulating infection structure differentiation and secondary metabolism of A. alternata.

Significance and impact of study

Our study provides a theoretical basis for further in-depth investigation of the specific role of AaCaM in the calcium signalling pathway underlying hydrophobic and pear wax-induced infection structure differentiation and pathogenicity of A. alternata.
Cardiac resynchronization therapy response in cardiac sarcoidosis
Authors: Shabtaie Samuel A. , Sehrawat Ojasav , Lee Justin Z. , Cha Yong‐Mei , Mulpuru Siva K. , Kowlgi Narayan G. , Siontis Konstantinos C. , Rosenbaum Andrew N. , Bois John P. , AbouEzzeddine Omar F. , Noseworthy Peter A. , Asirvatham Samuel J. , DeSimone Christopher V. , Deshmukh Abhishek J.
Published: July 2022

Introduction

Cardiac sarcoidosis (CS) is a nonischemic cardiomyopathy (NICM) characterized by infiltration of noncaseating granulomas involving the heart with highly variable clinical manifestations that can include conduction abnormalities and systolic heart failure. Cardiac resynchronization therapy (CRT) has shown significant promise in NICM, though little is known about its efficacy in patients with CS.

Objective

To determine if CRT improved cardiac remodeling in patients with CS.

Methods

We retrospectively reviewed all patients with a clinical or histological diagnosis of CS who underwent CRT implantation at the Mayo Clinic enterprise from 2000 to 2021. Baseline characteristics, imaging parameters, heart failure hospitalizations and need for advanced therapies, and major adverse cardiac events (MACE) were assessed.

Results

Our cohort was comprised of 55 patients with 61.8% male and a mean age of 58.7 ± 10.9 years. Eighteen (32.7%) patients had definite CS, 21 (38.2%) had probable CS, while 16 (29.1%) had presumed CS, and 26 (47.3%) with extracardiac sarcoidosis. The majority underwent CRT‐D implantation (n = 52, 94.5%) and 3 (5.5%) underwent CRT‐P implantation with 67.3% of implanted devices being upgrades from prior pacemakers or implantable cardioverter defibrillators. At 6 months postimplantation there was no significant improvement in ejection fraction (34.8 ± 10.9% vs. 37.7 ± 14.2%, p = .331) or left ventricular end‐diastolic diameter (58.5 ± 10.2 vs. 57.5 ± 8.1 mm, p = .236), though mild improvement in left ventricular end systolic diameter (49.1 ± 9.9 vs. 45.7± 9.9 mm, p < .0001). Within the first 6 months postimplantation, 5 (9.1%) patients sustained a heart failure hospitalization. At a mean follow‐up of 4.1± 3.7 years, 14 (25.5%) patients experienced a heart failure hospitalization, 11 (20.0%) underwent cardiac transplantation, 1 (1.8%) underwent left ventricular assist device implantation and 7 (12.7%) patients died.

Conclusions

Our findings suggest variable response to CRT in patients with CS with no overall improvement in ventricular function within 6 months and a substantial proportion of patients progressing to advanced heart failure therapies.
Impact of medication adherence on the efficacy of Baroreflex activation therapy
Authors: Schäfer Ann‐Kathrin C. , Müller Dieter , Born Ellen , Mühlhaus Maria , Lüders Stephan , Wallbach Manuel , Koziolek Michael J.
Published: July 2022

Therapy adherence significantly determines the success of antihypertensive therapy, especially in patients with resistant hypertension. Our study investigates the impact of drug adherence on the efficacy of Baroreflex‐activation‐therapy (BAT). In this retrospective analysis, the authors measured blood pressure (BP) and antihypertensive medication adherence (by gas chromatography‐mass spectrometry [GC‐MS] urine analysis) before and 6 months after BAT initiation. Adherence was defined as detection of ≥80% intake of prescribed medication at the time of follow‐up. Response to BAT was defined as BP drop ≥5 mmHg in systolic 24 h‐ambulatory BP (ABP) after 6 months. Overall patients (n = 38) median medication adherence was low, but rose from 60% (IQR 25%–100%) to 75% (IQR 38%–100%; p = .0194). After 6 months of BAT, mean systolic and diastolic office BP (‐21 ± 25 mmHg and ‐9 ± 15 mmHg; p < .0001 and .0004) as well as 24 h‐ABP dropped significantly (‐9 ± 17 mmHg and ‐5 ± 12 mmHg; p = .0049 and .0280). After 6 months of BAT, 21 patients (60%) could be classified as responders. There was neither significant difference in mean office systolic (‐21 ± 23 mmHg vs. ‐21 ± 28 mmHg; p = .9581) nor in 24 h‐systolic ABP decrease (‐11 ± 19 mmHg vs. ‐7 ± 15 mmHg; p = .4450) comparing adherent and non‐adherent patients. Whereas Antihypertensive Therapeutic Index (ATI) was unchanged in non‐responders, it significantly decreased in responders (from 50 ± 16 to 46 ± 16; p = .0477). These data are the first to show that BAT‐initiation leads to a clear BP reduction independently of patients´ medication adherence. Response to BAT is associated with a significant lowering of ATI, which might contribute to an underestimation of BAT efficacy.
Factors associated with hypoglycemia unawareness and severe hypoglycemia in type 1 diabetes mellitus patients
Authors: Takagi Satoshi , Miura Junnosuke , Takita Mikako , Mochizuki Shota , Asanuma Takuya , Hoshina Sari , Takaike Hiroko , Uchigata Yasuko , Babazono Tetsuya
Published: July 2022

Aims/Introduction

Several factors are associated with hypoglycemia unawareness and severe hypoglycemia, but few large studies have analyzed Japanese patients with type 1 diabetes. The aim of this study was to analyze the risk factors for hypoglycemia unawareness and severe hypoglycemia in Japanese type 1 diabetes patients.

Materials and Methods

A self‐administered questionnaire investigated events, complications and treatments associated with hypoglycemia in patients with type 1 diabetes. Multiple logistic regression analysis of factors associated with hypoglycemia unawareness and severe hypoglycemia requiring medical treatment was carried out. The coefficient of variation (CV) of blood glucose levels was determined using blood samples collected at six outpatient visits.

Results

Of the 1,619 participants, 44.2% and 10.4% experienced hypoglycemia unawareness and severe hypoglycemia, respectively. Mean HbA1c levels in patients with hypoglycemia unawareness were lower than those in patients without hypoglycemia unawareness. The type 1 diabetes subtype, glycated hemoglobin (HbA1c) level, CV of blood glucose levels and history of severe hypoglycemia requiring medical treatment were significant independent variables predicting the presence of hypoglycemia unawareness. The glucose CV and a history of hypoglycemia unawareness were significant independent variables predicting severe hypoglycemia requiring medical treatment. In stratified analyses of patients divided into four groups according to glucose CV and HbA1c levels, the high‐glucose‐CV/low‐HbA1c group had the highest odds ratios for hypoglycemia unawareness (2.60) and severe hypoglycemia requiring medical treatment (2.55).

Conclusions

The ambulant glucose CV correlated with both hypoglycemia unawareness and severe hypoglycemia. Patients with high glucose CV and low HbA1c are at high risk of such adverse events, and their treatment strategies should be reviewed.