bluuebunny/crossref_metadata_2025 huggingface dataset:
Attachment-related anxiety is associated with poor genital satisfaction and sexual problems in women
Authors: Komlenac Nikola , Hochleitner Margarethe
Published: November 2020

Background

Genital satisfaction has been found to influence women’s sexual experience. We tested the Relational Bodily Experiences Theory (RBET) that predicts associations between women’s genital satisfaction, attachment models, and sexual desire. We extended the model by additionally considering sexual arousal, orgasmic sensation, or the experience of pain during sexual activity as outcome variables. According to the RBET, women’s attachment models are associated with their genital satisfaction and linked to women’s sexual experience.

Methods

A cross-sectional online questionnaire study was conducted at an Austrian medical university. In total 294 women (Mage = 23.7, SD = 3.4) provided full responses. Women were asked about genital satisfaction and experiences of distressing sexual problems. Attachment-related anxiety and avoidance were assessed with the Experiences in Close Relationships-Relationship Structures Questionnaire.

Results

Results partially support the RBET. Attachment-related anxiety was associated with genital satisfaction which, in turn, was linked to experiences of frequent and/or distressing diminished sexual arousal, diminished sexual desire, or pain during sexual activity.

Conclusions

These results suggest that clinicians should assess genital satisfaction when treating female sexual problems. Women with attachment-related anxiety may especially be prone to having poor genital satisfaction and may profit from body image interventions in order to improve their sexual experience.
Descriptive study of how proportioning marks determines the performance of nursing students in a pharmacology course
Authors: Doggrell Sheila Anne
Published: November 2020

Background

In programs with higher proportions of marks allocated to ongoing assessment, the students have higher overall marks than those with a lower proportion allocated to assessment. Little or no attention has been made to how the allocation affects the academic success of students in individual courses. The purpose of this study was to determine how the allocation of marks to examinations, tutorials and an assignment affects the performance of nursing students in a pharmacology course.

Methods

For students who passed a pharmacology course (i) the marks for examinations and ongoing assessment (tutorials and/or an assignment) were compared, and (ii) regression line and correlation analysis was undertaken to determine any association between these marks. In addition, for completing students, modelling was undertaken to determine the effects of changing the allocation of marks on passing and failing rates.

Results

Nursing students who passed a pharmacology course obtained significantly lower marks in examinations than ongoing assessment, and for the ongoing assessment, lower marks in the assignment than tutorials. Regression line analysis showed that the marks in ongoing assessment (tutorials and/or the assignment) versus examination marks were a poor line-fit. The correlation coefficients between ongoing assessment and examinations were weak to moderate. A high percentage of students passed the course (> 90%) and, modelling for completing students, showed that decreasing the marks for examination would have led to slightly more students passing the pharmacology course with higher grades. In contrast, increasing the marks for examination would have dramatically decreased the number of students passing the course, and their grades.

Conclusions

The allocation of marks can have a major effect on student performance. As ongoing assessment is only a weak or moderate indicator of performance in examination this has implications for students who rely on passing examinations for their advancement. For instance, nursing students in some countries (e.g. USA) are required to pass examinations prior to registration, whereas in others (e.g. Australia) they are not. Consideration needs to be given as to whether it is appropriate for nursing students who fail examinations to pass courses/programs.
The patient at the centre: evidence from 17 European integrated care programmes for persons with complex needs
Authors: Czypionka Thomas , Kraus Markus , Reiss Miriam , Baltaxe Erik , Roca Josep , Ruths Sabine , Stokes Jonathan , Struckmann Verena , Haček Romana Tandara , Zemplényi Antal , Hoedemakers Maaike , Rutten-van Mölken Maureen
Published: November 2020

Background

As the prevalence of multi-morbidity increases in ageing societies, health and social care systems face the challenge of providing adequate care to persons with complex needs. Approaches that integrate care across sectors and disciplines have been increasingly developed and implemented in European countries in order to tackle this challenge. The aim of the article is to identify success factors and crucial elements in the process of integrated care delivery for persons with complex needs as seen from the practical perspective of the involved stakeholders (patients, professionals, informal caregivers, managers, initiators, payers).

Methods

Seventeen integrated care programmes for persons with complex needs in 8 European countries were investigated using a qualitative approach, namely thick description, based on semi-structured interviews and document analysis. In total, 233 face-to-face interviews were conducted with stakeholders of the programmes between March and September 2016. Meta-analysis of the individual thick description reports was performed with a focus on the process of care delivery.

Results

Four categories that emerged from the overarching analysis are discussed in the article: (1) a holistic view of the patient, considering both mental health and the social situation in addition to physical health, (2) continuity of care in the form of single contact points, alignment of services and good relationships between patients and professionals, (3) relationships between professionals built on trust and facilitated by continuous communication, and (4) patient involvement in goal-setting and decision-making, allowing patients to adapt to reorganised service delivery.

Conclusions

We were able to identify several key aspects for a well-functioning integrated care process for complex patients and how these are put into actual practice. The article sets itself apart from the existing literature by specifically focussing on the growing share of the population with complex care needs and by providing an analysis of actual processes and interpersonal relationships that shape integrated care in practice, incorporating evidence from a variety of programmes in several countries.
Education of medical personnel optimizes filling volume of blood culture bottles without negatively affecting microbiology testing
Authors: Steiner Katrin , Baron-Stefaniak Joanna , Hirschl Alexander M. , Barousch Wolfgang , Willinger Birgit , Baron David M.
Published: November 2020

Background

Anemia is a risk factor for adverse outcomes, which can be aggravated by unnecessary phlebotomies. In blood culture testing, up to 30 ml of blood can be withdrawn per sample, even though most manufacturers recommend blood volumes of 10 ml or less. After assessing the filling volume of blood culture bottles at our institution, we investigated whether an educational intervention could optimize filling volume of blood culture bottles without negatively affecting microbiology testing.

Methods

We weighed 10,147 blood cultures before and 11,806 blood cultures after a six-month educational intervention, during which employees were trained regarding correct filling volume via lectures, handouts, emails, and posters placed at strategic places.

Results

Before the educational intervention, only 31% of aerobic and 34% of anaerobic blood cultures were filled correctly with 5–10 ml of blood. The educational intervention increased the percentage of correctly filled bottles to 43% (P < 0.001) for both aerobic and anaerobic samples without negatively affecting results of microbiologic testing. In addition, sample volume was reduced from 11.0 ± 6.5 to 9.4 ± 5.1 ml (P < 0.001) in aerobic bottles and from 10.1 ± 5.6 to 8.8 ± 4.8 ml (P < 0.001) in anaerobic bottles.

Conclusion

Education of medical personnel is a simple and effective way to reduce iatrogenic blood loss and possibly moderate the extent of phlebotomy-induced anemia.
First evidence of convergent lifestyle signal in reptile skull roof microanatomy
Authors: Ebel Roy , Müller Johannes , Ramm Till , Hipsley Christy , Amson Eli
Published: November 2020

Background

The study of convergently acquired adaptations allows fundamental insight into life’s evolutionary history. Within lepidosaur reptiles—i.e. lizards, tuatara, and snakes—a fully fossorial (‘burrowing’) lifestyle has independently evolved in most major clades. However, despite their consistent use of the skull as a digging tool, cranial modifications common to all these lineages are yet to be found. In particular, bone microanatomy, although highly diagnostic for lifestyle, remains unexplored in the lepidosaur cranium. This constitutes a key gap in our understanding of their complexly interwoven ecology, morphology, and evolution. In order to bridge this gap, we reconstructed the acquisition of a fossorial lifestyle in 2813 lepidosaurs and assessed the skull roof compactness from microCT cross-sections in a representative subset (n = 99). We tested this and five macroscopic morphological traits for their convergent evolution.

Results

We found that fossoriality evolved independently in 54 lepidosaur lineages. Furthermore, a highly compact skull roof, small skull diameter, elongate cranium, and low length ratio of frontal and parietal were repeatedly acquired in concert with a fossorial lifestyle.

Conclusions

We report a novel case of convergence that concerns lepidosaur diversity as a whole. Our findings further indicate an early evolution of fossorial modifications in the amphisbaenian ‘worm-lizards’ and support a fossorial origin for snakes. Nonetheless, our results suggest distinct evolutionary pathways between fossorial lizards and snakes through different contingencies. We thus provide novel insights into the evolutionary mechanisms and constraints underlying amniote diversity and a powerful tool for the reconstruction of extinct reptile ecology.
FFAR1 activation attenuates histamine-induced myosin light chain phosphorylation and cortical tension development in human airway smooth muscle cells
Authors: Xu Shengjie , Schwab Anthony , Karmacharya Nikhil , Cao Gaoyuan , Woo Joanna , Kim Nicholas , An Steven S. , Panettieri Jr Reynold A. , Jude Joseph A.
Published: November 2020

Background

Activation of free fatty acid receptors (FFAR1 and FFAR4) which are G protein-coupled receptors (GPCRs) with established (patho)physiological roles in a variety of obesity-related disorders, induce human airway smooth muscle (HASM) cell proliferation and shortening. We reported amplified agonist-induced cell shortening in HASM cells obtained from obese lung donors. We hypothesized that FFAR1 modulate excitation–contraction (EC) coupling in HASM cells and play a role in obesity-associated airway hyperresponsiveness.

Methods

In HASM cells pre-treated (30 min) with FFAR1 agonists TAK875 and GW9508, we measured histamine-induced Ca2+ mobilization, myosin light chain (MLC) phosphorylation, and cortical tension development with magnetic twisting cytometry (MTC). Phosphorylation of MLC phosphatase and Akt also were determined in the presence of the FFAR1 agonists or vehicle. In addition, the effects of TAK875 on MLC phosphorylation were measured in HASM cells desensitized to β2AR agonists by overnight salmeterol treatment. The inhibitory effect of TAK875 on MLC phosphorylation was compared between HASM cells from age and sex-matched non-obese and obese human lung donors. The mean measurements were compared using One-Way ANOVA with Dunnett’s test for multiple group comparisons or Student’s t-test two-group comparison. For cortical tension measurements by magnetic twisted cytometry, mixed effect model using SAS V.9.2 was applied. Means were considered significant when p ≤ 0.05.

Results

Unexpectedly, we found that TAK875, a synthetic FFAR1 agonist, attenuated histamine-induced MLC phosphorylation and cortical tension development in HASM cells. These physiological outcomes were unassociated with changes in histamine-evoked Ca2+ flux, protein kinase B (AKT) activation, or MLC phosphatase inhibition. Of note, TAK875-mediated inhibition of MLC phosphorylation was maintained in β2AR-desensitized HASM cells and across obese and non-obese donor-derived HASM cells.

Conclusions

Taken together, our findings identified the FFAR1 agonist TAK875 as a novel bronchoprotective agent that warrants further investigation to treat difficult-to-control asthma and/or airway hyperreactivity in obesity.
Effect of diet and intestinal AhR expression on fecal microbiome and metabolomic profiles
Authors: Yang Fang , DeLuca Jennifer A. A. , Menon Rani , Garcia-Vilarato Erika , Callaway Evelyn , Landrock Kerstin K. , Lee Kyongbum , Safe Stephen H. , Chapkin Robert S. , Allred Clinton D. , Jayaraman Arul
Published: November 2020

Background

Diet, loss of aryl hydrocarbon receptor (AhR) expression and their modification of the gut microbiota community composition and its metabolites affect the development of colorectal cancer (CRC). However, the concordance between fecal microbiota composition and the fecal metabolome is poorly understood. Mice with specific AhR deletion (AhRKO) in intestinal epithelial cell and their wild-type littermates were fed a low-fat diet or a high-fat diet. Shifts in the fecal microbiome and metabolome associated with diet and loss of AhR expression were assessed. Microbiome and metabolome data were integrated to identify specific microbial taxa that contributed to the observed metabolite shifts.

Results

Our analysis shows that diet has a more pronounced effect on mouse fecal microbiota composition than the impact of the loss of AhR. In contrast, metabolomic analysis showed that the loss of AhR in intestinal epithelial cells had a more pronounced effect on metabolite profile compared to diet. Integration analysis of microbiome and metabolome identified unclassified Clostridiales, unclassifiedDesulfovibrionaceae, andAkkermansiaas key contributors to the synthesis and/or utilization of tryptophan metabolites.

Conclusions

Akkermansiaare likely to contribute to the synthesis and/or degradation of tryptophan metabolites. Our study highlights the use of multi-omic analysis to investigate the relationship between the microbiome and metabolome and identifies possible taxa that can be targeted to manipulate the microbiome for CRC treatment.
The generation of metabolic changes for the production of high-purity zeaxanthin mediated by CRISPR-Cas9 in Chlamydomonas reinhardtii
Authors: Song Inhwa , Kim Jongrae , Baek Kwangryul , Choi Young , Shin ByongCheol , Jin EonSeon
Published: November 2020

Background

Zeaxanthin, a major xanthophyll pigment, has a significant role as a retinal pigment and antioxidant. Because zeaxanthin helps to prevent age-related macular degeneration, its commercial use in personalized nutritional and pharmaceutical applications has expanded. To meet the quantitative requirements for personalized treatment and pharmaceutical applications, it is necessary to produce highly purified zeaxanthin.

Results

In this study, to meet the quantitative requirements for industrial applications, we generated a double knockout mutant which is gene-edited by the CRISPR-Cas9 ribonucleoprotein-mediated knock-in system. The lycopene epsilon cyclase (LCYE) was edited to the elimination of α-branch of xanthophyll biosynthesis in a knockout mutant of the zeaxanthin epoxidase gene (ZEP). The double knockout mutant (dzl) had a 60% higher zeaxanthin yield (5.24 mg L− 1) and content (7.28 mg g− 1) than that of the parental line after 3 days of cultivation. Furthermore, medium optimization improved the 3-day yield of zeaxanthin from the dzl mutant to 6.84 mg L− 1.

Conclusions

A Chlamydomonas strain with the elimination of lutein production by gene editing using CRISPR-Cas9 has been successfully developed. This research presents a solution to overcome the difficulties of the downstream-process for the production of high-purity zeaxanthin.
Reporting on the opioid crisis (2000–2018): role of The Globe and Mail, a Canadian English-language newspaper in influencing public opinion
Authors: Quan Amanda My Linh , Wilson Lindsay A. , Mithani Salima S. , Zhu David T. , Bota A. Brianne , Wilson Kumanan
Published: November 2020

Background

We aim to describe the general characteristics of how the Canadian newspaper The Globe and Mail reports on opioid-related news, the opioid crisis and its victims, and explore how Canadians’ perceptions of the opioid crisis could have developed over time from this reporting. The Globe and Mail has the highest circulation among Canadian newspapers and is Canada’s newspaper of record.

Methods

Reviewers performed independent, blinded bibliometric searches of all The Globe and Mail articles archived in the Canadian Periodicals Index Quarterly spanning an 18-year period (1 January 2000–1 June 2018) related to the keywords “opioids” or “drugs and opioids” and “opiates”. Independently and in duplicate, reviewers manually extracted qualitative data from articles and identified emergent themes. Articles were screened independently by both reviewers based on the inclusion criteria. Conflicts were resolved by discussion and consensus. Social representation theory was used as a framework for describing how the opioid crisis is portrayed in Canada.

Results

Our search yielded 650 relevant opioid articles. The number of articles peaked in 2009, 2012, and in 2016, coinciding with major developments in the epidemic. The language used in this discourse has evolved over the years and has slowly shifted towards less stigmatizing language. Content analysis of the articles revealed common social representations attributing responsibility to pharmaceutical companies, physicians, and foreign countries.

Conclusions

The Globe and Mail’s coverage of the opioid crisis is focused on basic social representations and attributed responsibility for the crisis to a few collectives. A shift toward coverage of the root causes of the opioid epidemic could positively influence the general public’s perception of the opioid crisis and promote deeper understanding of the issue. Journalists face several obstacles to achieve greater focus and framing of the opioid crisis; a closer working relationship between the media and the research community is needed.
Rasch analysis of the long-term conditions questionnaire (LTCQ) and development of a short-form (LTCQ-8)
Authors: Batchelder Laurie , Fox Diane , Potter Caroline M. , Peters Michele , Jones Karen , Forder Julien E. , Fitzpatrick Ray
Published: November 2020

Background

The aim of the current study was to evaluate the structural validity of the 20-item long-term conditions questionnaire (LTCQ) and to explore a potential short-form version of the scale using Rasch analysis.

Methods

Data were collected through postal surveys (February 2016–January 2017) from a sample of 1,211 participants diagnosed with at least one long-term condition (LTC). Identified participants were invited through either local authorities for a social care cohort (n = 294) or primary care practices for a health care cohort (n = 917). Participants were mailed a survey, including the LTCQ, demographic questions, a comorbidities measure, and other validated outcome measures. Respondents were invited to complete a follow-up survey including the LTCQ for assessment of reproducibility.

Results

The main assumptions of the Rasch model from the LTCQ were fulfilled, although infit and outfit indices indicated some items showed misfit. Misfitted items, items that did not have a preceding set or showed some local dependence were removed one at a time, with the remaining candidate items to form an 8-item short version, the LTCQ-8. The Rasch model for the LTCQ-8 explained 64% variance and had a reliability estimate greater than 0.80. Several items in the LTCQ showed uniform differential item function (DIF) in relation to the number of reported LTCs, age, cohort and type of LTCs, but fewer items exhibited DIF in the LTCQ-8. Spearman’s rho correlations between the LTCQ and the LTCQ-8 were strong across the total sample and various subgroups. Correlations between the LTCQ-8 and all reference measures were moderate to strong, and comparable to correlations found between the LTCQ and these measures.

Conclusions

The LTCQ measures a unidimensional construct, and it is therefore acceptable to use a summed total score. The LTCQ-8 also met the assumption of unidimensionality and had comparable construct validity with the LTCQ. Additional validation is required in an independent sample.
The association of alcohol with circulating total fibrinogen and plasma clot density is mediated by fibrinogen and FXIII genotypes
Authors: Rautenbach Petro Hannie , Nienaber-Rousseau Cornelie , Pieters Marlien
Published: November 2020

Background

Alcohol consumption is associated with haemostasis and so may influence cardiovascular conditions. It is unknown whether the association of alcohol with total and γ’ fibrinogen concentrations, as well as clot structure, are modulated by fibrinogen andfactor (F) XIIIsingle nucleotide polymorphisms (SNPs).

Methods

Total fibrinogen, γ’ fibrinogen and clot properties of 2010 healthy Africans residing in South Africa were measured in relation to alcohol intake as well as its markers – gamma-glutamyltransferase (GGT), percentage carbohydrate deficient transferrin (%CDT), aspartate aminotransferase (AST), and alanine aminotransferase (ALT). Fourteenfibrinogenand two SNPs in theFXIIIgene were genotyped to determine their influence.

Results

Alcohol intake and its markers correlated negatively with fibrinogen and clot lysis time (CLT) as well as with most of the clot properties. Percentage γ’ fibrinogen correlated positively with AST and negatively with alcohol intake. We then stratified for alcohol intake and found inverse associations between γ’ fibrinogen and both %CDT and GGT–CDT in consumers, but the positive association with AST remained only in abstainers. Alcohol intake and its markers modulated the influence offibrinogenSNPs on total fibrinogen concentrations and thefibrinogenSNPs as well as anFXIIISNP on clot density (allp < 0.004).

Conclusion/s

We show for the first time that some individuals harbour certain genotypes that, in combination with alcohol consumption, might predispose or protect them from haemostatic factors that might lead to the development of cardiovascular disease. Studies are needed to clarify the mechanisms related to the interplay between alcohol and the gene variants observed here.
Maternal mortality following thromboembolism; incidences and prophylaxis strategies
Authors: Shirazi Mahboobeh , Sahebdel Behrokh , Torkzaban Mahnoosh , Feizabad Elham , Ghaemi Marjan
Published: November 2020

Background

Thromboembolism is one of the main causes of maternal mortality, which can be prevented in many cases. The present study was designed to investigate the incidence and prophylaxis strategies for maternal mortality following thromboembolism in postnatal.

Methods

In this case series study, the data of the mortality cases were extracted according to the ethical and security standards of the Ministry of Health of the country and compared with a healthy control group. The thromboembolism risk factors measured and scored using a questionnaire entitled “the evaluation of risk factors for maternal mortality following thromboembolism during pregnancy, labor, or post-partum”.

Results

The maternal mortality rate was 16 per 100,000 live births. Among 297 mortality cases, 27 (9%) death were due to thromboembolism. The mean gestational age was 32.5 weeks. Dyspnea (88.8%) and tachycardia (18.5%) were found as common clinical manifestations in these patients. Sixteen cases (59.3%) did not get heparin, 6 (22.2%) received single dose and 5 (18.5%) received two doses and more. In these 11 cases, 5 (45%) patients received heparin before surgery, 1 after surgery, and 5 before and after surgery. Twenty cases deceased in the first hours after delivery and the rest after 2 to 12 days. The average score of risk for thromboembolism based on Royal College of Obstetricians & Gynecologist (RCOG) guideline was 4.6.

Conclusion

It seems that one of the most important cause of maternal mortality in this study was the lack of recognition of high-risk patients and the lack of prescription for prophylaxis with heparin and this clearly explains the need for accurate screening of high-risk mothers, designing a standard form and the care and treatment of these patients.
Pathogenic effects of inhibition of mTORC1/STAT3 axis facilitates Staphylococcus aureus-induced pyroptosis in human macrophages
Authors: Yao Ruiyuan , Chen Yuhao , Hao Huifang , Guo Zhixin , Cheng Xiaoou , Ma Yuze , Ji Qiang , Yang Xiaoru , Wang Yanfeng , Li Xihe , Wang Zhigang
Published: November 2020

Background

Pyroptosis is a recently identified pathway of caspase-mediated cell death in response to microbes, lipopolysaccharide, or chemotherapy in certain types of cells. However, the mechanism of how pyroptosis is regulated is not well-established.

Methods

Herein, the intracellular bacteria were detected by staining and laser confocal microscopy and TEM. Live/dead cell imaging assay was used to examine macrophage death. Western blot and immunohistochemical staining were used to examine the protein changes. IFA was used to identify typical budding vesicles of pyroptosis and the STAT3 nuclear localization. SEM was used to observe the morphological characteristics of pyroptosis. ELISA was used to detect the level of inflammatory cytokines. Pyroptosis was filmed in macrophages by LSCM.

Results

S. aureus was internalized by human macrophages. Intracellular S. aureus induced macrophage death. S. aureus invasion increased the expression of NLRP3, Caspase1 (Casp-1 p20) and the accumulation of GSDMD-NT, GSDMD-NT pore structures, and the release of IL-1β and IL-18 in macrophages. Macrophages pyroptosis induced by S. aureus can be abrogated by blockage of S. aureus phagocytosis. The pyroptosic effect by S. aureus infection was promoted by either rapamycin or Stattic, a specific inhibitor for mTORC1 or STAT3. Inhibition of mTORC1 or STAT3 induced pyroptosis. mTORC1 regulated the pyroptosic gene expression through governing the nuclear localization of STAT3. mTORC1/STAT3 axis may play a regulatory role in pyroptosis within macrophages.

Conclusions

S. aureus infection induces human macrophage pyroptosis, inhibition of mTORC1/STAT3 axis facilitates S. aureus-induced pyroptosis. mTORC1 and STAT3 are associated with pyroptosis. Our findings demonstrate a regulatory function of the mTORC1/STAT3 axis in macrophage pyroptosis, constituting a novel mechanism by which pyroptosis is regulated in macrophages.
Heparanase and the hallmarks of cancer
Authors: Jayatilleke Krishnath M. , Hulett Mark D.
Published: November 2020

Heparanase is the only mammalian enzyme that cleaves heparan sulphate, an important component of the extracellular matrix. This leads to the remodelling of the extracellular matrix, whilst liberating growth factors and cytokines bound to heparan sulphate. This in turn promotes both physiological and pathological processes such as angiogenesis, immune cell migration, inflammation, wound healing and metastasis. Furthermore, heparanase exhibits non-enzymatic actions in cell signalling and in regulating gene expression. Cancer is underpinned by key characteristic features that promote malignant growth and disease progression, collectively termed the ‘hallmarks of cancer’. Essentially, all cancers examined to date have been reported to overexpress heparanase, leading to enhanced tumour growth and metastasis with concomitant poor patient survival. With its multiple roles within the tumour microenvironment, heparanase has been demonstrated to regulate each of these hallmark features, in turn highlighting the need for heparanase-targeted therapies. However, recent discoveries which demonstrated that heparanase can also regulate vital anti-tumour mechanisms have cast doubt on this approach. This review will explore the myriad ways by which heparanase functions as a key regulator of the hallmarks of cancer and will highlight its role as a major component within the tumour microenvironment. The dual role of heparanase within the tumour microenvironment, however, emphasises the need for further investigation into defining its precise mechanism of action in different cancer settings.
Logistics of an advanced therapy medicinal product during COVID-19 pandemic in Italy: successful delivery of mesenchymal stromal cells in dry ice
Authors: Astori Giuseppe , Bernardi Martina , Bozza Angela , Catanzaro Daniela , Chieregato Katia , Merlo Anna , Santimaria Monica , Barbazza Roberto , Amodeo Giuseppe , Ciccocioppo Rachele , Elice Francesca , Ruggeri Marco
Published: November 2020

Background

During the coronavirus disease-2019 (COVID-19) pandemic, Italian hospitals faced the most daunting challenges of their recent history, and only essential therapeutic interventions were feasible. From March to April 2020, the Laboratory of Advanced Cellular Therapies (Vicenza, Italy) received requests to treat a patient with severe COVID-19 and a patient with acute graft-versus-host disease with umbilical cord-derived mesenchymal stromal cells (UC-MSCs). Access to clinics was restricted due to the risk of contagion. Transport of UC-MSCs in liquid nitrogen was unmanageable, leaving shipment in dry ice as the only option.

Methods

We assessed effects of the transition from liquid nitrogen to dry ice on cell viability; apoptosis; phenotype; proliferation; immunomodulation; and clonogenesis; and validated dry ice-based transport of UC-MSCs to clinics.

Results

Our results showed no differences in cell functionality related to the two storage conditions, and demonstrated the preservation of immunomodulatory and clonogenic potentials in dry ice. UC-MSCs were successfully delivered to points-of-care, enabling favourable clinical outcomes.

Conclusions

This experience underscores the flexibility of a public cell factory in its adaptation of the logistics of an advanced therapy medicinal product during a public health crisis. Alternative supply chains should be evaluated for other cell products to guarantee delivery during catastrophes.
Association between metabolic syndrome and venous thromboembolism after total joint arthroplasty: a meta-analysis of cohort studies
Authors: Yang Yipei , Li Ziyue , Liang Haifeng , Tian Jing
Published: November 2020

Objective

Metabolic syndrome (MetS) has been associated with hypercoagulative status. However, previous studies evaluating the association between MetS and incidence of venous thromboembolism (VTE) after total joint arthroplasty (TJA) showed inconsistent results. We performed a meta-analysis to evaluate the influence of MetS on the risk of VTE following TJA.

Methods

Cohort studies were identified by the search of PubMed, Embase, and the Cochrane’s Library databases. A random-effect model was used if considerable heterogeneity was detected; otherwise, a fixed-effect model was used. Subgroup analyses according to the category of VTE, definition of MetS, category of procedure, and follow-up durations were performed.

Results

Seven cohort studies with 1,341,457 patients that underwent TJA were included, with 118,060 MetS patients (8.8%) at baseline. With a follow-up duration up to 3 months after surgery, 9788 patients had VTE. Pooled results with a random-effect model showed that MetS was not associated with increased overall VTE after TJA (adjusted risk ratio [RR] = 1.24, 95% confidence interval [CI] 0.89 ~ 1.72, p = 0.20; I2 = 69%). The results were not significantly affected by the diagnostic criteria of MetS, category of the procedure, and follow-up durations. Subgroup analyses showed that MetS was not associated with an increased the risk of pulmonary embolism ([PE], RR 1.06, 95% CI 0.37 ~ 3.02, p = 0.91), but an increased risk of deep vein thrombosis (DVT) after TJA (RR 3.38, 95% CI 1.83 ~ 6.24, p < 0.001).

Conclusions

Current evidence from observational studies suggests MetS might be associated with an increased risk of DVT but not PE after TJA.
A 29 Mainland Chinese cohort of patients with Phelan–McDermid syndrome: genotype–phenotype correlations and the role of SHANK3 haploinsufficiency in the important phenotypes
Authors: Xu Na , Lv Hui , Yang Tingting , Du Xiujuan , Sun Yu , Xiao Bing , Fan Yanjie , Luo Xiaomei , Zhan Yongkun , Wang Lili , Li Fei , Yu Yongguo
Published: November 2020

Background

Phelan–McDermid syndrome (PMS) or 22q13 deletion syndrome is a rare developmental disorder characterized by hypotonia, developmental delay (DD), intellectual disability (ID), autism spectrum disorder (ASD) and dysmorphic features. Most cases are caused by 22q13 deletions encompassing many genes including SHANK3. Phenotype comparisons between patients with SHANK3 mutations (or deletions only disrupt SHANK3) and 22q13 deletions encompassing more than SHANK3 gene are lacking.

Methods

A total of 29 Mainland China patients were clinically and genetically evaluated. Data were obtained from medical record review and a standardized medical history questionnaire, and dysmorphology evaluation was conducted via photographic evaluation. We analyzed 22q13 deletions and SHANK3 small mutations and performed genotype–phenotype analysis to determine whether neurological features and other important clinical features are responsible for haploinsufficiency of SHANK3.

Results

Nineteen patients with 22q13.3 deletions ranging in size from 34 kb to 8.7 Mb, one patient with terminal deletions and duplications, and nine patients with SHANK3 mutations were included. All mutations would cause loss-of function effect and six novel heterozygous variants, c.3838_3839insGG, c.3088delC, c.3526G > T, c.3372dupC, c.3120delC and c.3942delC, were firstly reported. Besides, we demonstrated speech delay (100%), DD/ID (88%), ASD (80%), hypotonia (83%) and hyperactivity (83%) were prominent clinical features. Finally, 100% of cases with monogenic SHANK3 deletion had hypotonia and there was no significant difference between loss of SHANK3 alone and deletions encompassing more than SHANK3 gene in the prevalence of hypotonia, DD/ID, ASD, increased pain tolerance, gait abnormalities, impulsiveness, repetitive behaviors, regression and nonstop crying which were high in loss of SHANK3 alone group.

Conclusions

This is the first work describing a cohort of Mainland China patients broaden the clinical and molecular spectrum of PMS. Our findings support the effect of 22q13 deletions and SHANK3 point mutations on language impairment and several clinical manifestations, such as DD/ID. We also demonstrated SHANK3 haploinsufficiency was a major contributor to the neurological phenotypes of PMS and also responsible for other important phenotypes such as hypotonia, increased pain tolerance, impulsiveness, repetitive behaviors, regression and nonstop crying.
Targeting the myeloid checkpoint receptor SIRPα potentiates innate and adaptive immune responses to promote anti-tumor activity
Authors: Kuo Tracy C. , Chen Amy , Harrabi Ons , Sockolosky Jonathan T. , Zhang Anli , Sangalang Emma , Doyle Laura V. , Kauder Steven E. , Fontaine Danielle , Bollini Sangeetha , Han Bora , Fu Yang-Xin , Sim Janet , Pons Jaume , Wan Hong I.
Published: November 2020

Background

Signal regulatory protein α (SIRPα) is a myeloid-lineage inhibitory receptor that restricts innate immunity through engagement of its cell surface ligand CD47. Blockade of the CD47–SIRPα interaction synergizes with tumor-specific antibodies and T-cell checkpoint inhibitors by promoting myeloid-mediated antitumor functions leading to the induction of adaptive immunity. Inhibition of the CD47–SIRPα interaction has focused predominantly on targeting CD47, which is expressed ubiquitously and contributes to the accelerated blood clearance of anti-CD47 therapeutics. Targeting SIRPα, which is myeloid-restricted, may provide a differential pharmacokinetic, safety, and efficacy profile; however, SIRPα polymorphisms and lack of pan-allelic and species cross-reactive agents have limited the clinical translation of antibodies against SIRPα. Here, we report the development of humanized AB21 (hAB21), a pan-allelic anti-SIRPα antibody that binds human, cynomolgus monkey, and mouse SIRPα alleles with high affinity and blocks the interaction with CD47.

Methods

Human macrophages derived from donors with various SIRPα v1 and v2 allelic status were used to assess the ability of hAB21 to enhance phagocytosis. HAB21_IgG subclasses were evaluated for targeted depletion of peripheral blood mononuclear cells, phagocytosis and in vivo efficacy in xenograft models. Combination therapy with anti-PD1/anti-PD-L1 in several syngeneic models was performed. Immunophenotyping of tissues from MC38 tumor-bearing mice treated with AB21 and anti-PD-1 was evaluated. PK, PD and tolerability of hAB21 were evaluated in cynomolgus monkeys.

Results

SIRPα blockade with hAB21 promoted macrophage-mediated antibody-dependent phagocytosis of tumor cells in vitro and improved responses to rituximab in the Raji human tumor xenograft mouse model. Combined with PD-1/PD-L1 blockade, AB21 improved response rates by facilitating monocyte activation, dendritic cell activation, and T cell effector functions resulting in long term, durable antitumor immunity. In cynomolgus monkeys, hAB21 has a half-life of 5.3 days at 10 mg/kg and complete target occupancy with no hematological toxicity or adverse findings at doses up to 30 mg/kg.

Conclusions

The in vitro and in vivo antitumor activity of hAB21 broadly recapitulates that of CD47 targeted therapies despite differences in ligand expression, binding partners, and function, validating the CD47–SIRPα axis as a fundamental myeloid checkpoint pathway and its blockade as promising therapeutic intervention for treatment of human malignancies.
Engineering crops of the future: CRISPR approaches to develop climate-resilient and disease-resistant plants
Authors: Zaidi Syed Shan-e-Ali , Mahas Ahmed , Vanderschuren Hervé , Mahfouz Magdy M.
Published: November 2020

To meet increasing global food demand, breeders and scientists aim to improve the yield and quality of major food crops. Plant diseases threaten food security and are expected to increase because of climate change. CRISPR genome-editing technology opens new opportunities to engineer disease resistance traits. With precise genome engineering and transgene-free applications, CRISPR is expected to resolve the major challenges to crop improvement. Here, we discuss the latest developments in CRISPR technologies for engineering resistance to viruses, bacteria, fungi, and pests. We conclude by highlighting current concerns and gaps in technology, as well as outstanding questions for future research.
Comparative effectiveness of direct admission and admission through emergency departments for children: a randomized stepped wedge study protocol
Authors: Leyenaar JoAnna K. , McDaniel Corrie E. , Acquilano Stephanie C. , Schaefer Andrew P. , Bruce Martha L. , O’Malley A. James
Published: November 2020

Background

Approximately 2 million children are hospitalized each year in the United States, with more than three-quarters of non-elective hospitalizations admitted through emergency departments (EDs). Direct admission, defined as admission to hospital without first receiving care in the hospital’s ED, may offer benefits for patients and healthcare systems in quality, timeliness, and experience of care. While ED utilization patterns are well studied, there is a paucity of research comparing the effectiveness of direct and ED admissions. The overall aim of this project is to compare the effectiveness of a standardized direct admission approach to admission beginning in the ED for hospitalized children.

Methods/design

We will conduct a stepped wedge cluster randomized controlled trial at 3 structurally and geographically diverse hospitals. A total of 70 primary and urgent care practice sites in the hospitals’ catchment areas will be randomized to a time point when they will begin participation in the multi-stakeholder informed direct admission program. This crossover will be unidirectional and occur at 4 time points, 6 months apart, over a 24-month implementation period. Our primary outcome will be the timeliness of clinical care provision. Secondary outcomes include (i) parent-reported experience of care, (ii) unanticipated transfer to the intensive care unit within 6 h of hospital admission, and (iii) rapid response calls within 6 h of hospital admission. We anticipate that 190 children and adolescents will be directly admitted, with 1506 admitted through EDs.

Analyses will compare the effectiveness of direct admission to admission through the ED and will evaluate the causal effect of implementing a direct admission program using linear regression with random effects for referring practice clusters and time period fixed effects. We will further examine the heterogeneity of treatment effects based on hypotheses specified a priori. In addition, we will conduct a mixed-methods process evaluation to assess reach, effectiveness, adoption, implementation, and maintenance of our direct admission intervention.

Discussion

Our study represents the first randomized controlled trial to compare the effectiveness of direct admission to admission through the ED for pediatric patients. Our scientific approach, pairing a stepped wedge design with a multi-level assessment of barriers to and facilitators of implementation, will generate valuable data about how positive findings can be reproduced across other healthcare systems.

Trial registration

ClinicalTrials.gov NCT04192799. Registered on December 10, 2019).
Characterization of an AA9 LPMO from Thielavia australiensis, TausLPMO9B, under industrially relevant lignocellulose saccharification conditions
Authors: Calderaro F. , Keser M. , Akeroyd M. , Bevers L. E. , Eijsink V. G. H. , Várnai A. , van den Berg M. A.
Published: November 2020

Background

The discovery of lytic polysaccharide monooxygenases (LPMO) has changed our perspective on enzymatic degradation of plant biomass. Through an oxidative mechanism, these enzymes are able to cleave and depolymerize various polysaccharides, acting not only on crystalline substrates such as chitin and cellulose, but also on other polysaccharides, such as xyloglucan, glucomannan and starch. Despite their widespread use, uncertainties related to substrate specificity and stereospecificity, the nature of the co-substrate, in-process stability, and the nature of the optimal reductant challenge their exploitation in biomass processing applications.

Results

In this work, we studied the properties of a novel fungal LPMO from the thermophilic fungusThielavia australiensis, TausLPMO9B. Heterologous expression ofTausLPMO9B inAspergillus nigeryielded a glycosylated protein with a methylated N-terminal histidine showing LPMO activity. High sequence identity of the AA9 domain to that ofMtLPMO9B (MYCTH_80312) fromMyceliophthora thermophila(84%) indicated strictly C1-oxidizing activity on cellulose, which was confirmed experimentally by the analysis of products released from cellulose using HPAEC. The enzyme was stable and active at a pH ranging from 4 to 6, thus matching the conditions commonly used in industrial biomass processing, where a low pH (between 4 and 5) is used due to the pH-optima of commercial cellulases and a desire to limit microbial contamination.

Conclusion

While the oxidative cleavage of phosphoric acid swollen cellulose (PASC) byTausLPMO9B was boosted by the addition of H2O2as a co-substrate, this effect was not observed during the saccharification of acid pretreated corn stover. This illustrates key differences between the lab-scale tests with artificial, lignin-free substrates and industrial settings with lignocellulosic biomass as substrate.
Comparative transcriptomic analysis of antimony resistant and susceptible Leishmania infantum lines
Authors: Andrade Juvana Moreira , Gonçalves Leilane Oliveira , Liarte Daniel Barbosa , Lima Davi Alvarenga , Guimarães Frederico Gonçalves , de Melo Resende Daniela , Santi Ana Maria Murta , de Oliveira Luciana Marcia , Velloso João Paulo Linhares , Delfino Renato Guimarães , Pescher Pascale , Späth Gerald F. , Ruiz Jeronimo Conceição , Murta Silvane Maria Fonseca
Published: November 2020

Background

One of the major challenges to leishmaniasis treatment is the emergence of parasites resistant to antimony. To study differentially expressed genes associated with drug resistance, we performed a comparative transcriptomic analysis between wild-type and potassium antimonyl tartrate (SbIII)-resistant Leishmania infantum lines using high-throughput RNA sequencing.

Methods

All the cDNA libraries were constructed from promastigote forms of each line, sequenced and analyzed using STAR for mapping the reads against the reference genome (L. infantum JPCM5) and DESeq2 for differential expression statistical analyses. All the genes were functionally annotated using sequence similarity search.

Results

The analytical pipeline considering an adjusted p-value < 0.05 and fold change > 2.0 identified 933 transcripts differentially expressed (DE) between wild-type and SbIII-resistant L. infantum lines. Out of 933 DE transcripts, 504 presented functional annotation and 429 were assigned as hypothetical proteins. A total of 837 transcripts were upregulated and 96 were downregulated in the SbIII-resistant L. infantum line. Using this DE dataset, the proteins were further grouped in functional classes according to the gene ontology database. The functional enrichment analysis for biological processes showed that the upregulated transcripts in the SbIII-resistant line are associated with protein phosphorylation, microtubule-based movement, ubiquitination, host–parasite interaction, cellular process and other categories. The downregulated transcripts in the SbIII-resistant line are assigned in the GO categories: ribonucleoprotein complex, ribosome biogenesis, rRNA processing, nucleosome assembly and translation.

Conclusions

The transcriptomic profile of L. infantum showed a robust set of genes from different metabolic pathways associated with the antimony resistance phenotype in this parasite. Our results address the complex and multifactorial antimony resistance mechanisms in Leishmania, identifying several candidate genes that may be further evaluated as molecular targets for chemotherapy of leishmaniasis.

Graphical Abstract