bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ribozyme-mediated CRISPR/Cas9 gene editing in pyrethrum (Tanacetum cinerariifolium) hairy roots using a RNA polymerase II-dependent promoter
Authors: Li Jia-Wen , Zeng Tuo , Xu Zhi-Zhuo , Li Jin-Jin , Hu Hao , Yu Qin , Zhou Li , Zheng Ri-Ru , Luo Jing , Wang Cai-Yun 
Published: March 2022

Abstract
Background
Traditional CRISPR/Cas9 systems that rely on U6 or U3 snRNA promoters (RNA polymerase III-dependent promoters) can only achieve constitutive gene editing in plants, hampering the functional analysis of specifically expressed genes. Ribozyme-mediated CRISPR/Cas9 systems increase the types of promoters which can be used to transcribe sgRNA. Therefore, such systems allow specific gene editing; for example, transcription of the artificial gene Ribozyme-sgRNA-Ribozyme (RGR) is initiated by an RNA polymerase II-dependent promoter. Genetic transformation is indispensable for editing plant genes. In certain plant species, including pyrethrum, genetic transformation remains challenging to do, limiting the functional verification of novel CRISPR/Cas9 systems. Thus, this study’s aim was to develop a simple Agrobacterium rhizogenes-mediated hairy root transformation system to analyze the function of a ribozyme-mediated CRISPR/Cas9 system in pyrethrum.

Results
A hairy root transformation system for pyrethrum is described, with a mean transformation frequency of 7%. Transgenic hairy roots transformed with the pBI121 vector exhibited significantly increased beta-glucuronidase staining as a visual marker of transgene expression. Further, a ribozyme-based CRISPR/Cas9 vector was constructed to edit the TcEbFS gene, which catalyzes synthesis of the defense-related compound (E)-β-farnesene in pyrethrum. The vector was transferred into the hairy roots of pyrethrum and two stably transformed hairy root transgenic lines obtained. Editing of the TcEbFS gene in the hairy roots was evaluated by gene sequencing, demonstrating that both hairy root transgenic lines had DNA base loss at the editing target site. Gas chromatography–mass spectrometry showed that the (E)-β-farnesene content was significantly decreased in both hairy root transgenic lines compared with the empty vector control group. Altogether, these results show that RGR can be driven by the CaMV35S promoter to realize TcEbFS gene editing in pyrethrum hairy roots.

Conclusion
An A. rhizogenes-mediated hairy root transformation and ribozyme-mediated CRISPR/Cas9 gene editing system in pyrethrum was established, thereby facilitating gene editing in specific organs or at a particular developmental stage in future pyrethrum research.
It’s time for a minimum synoptic operation template in patients undergoing laparoscopic cholecystectomy: a systematic review
Authors: O’Connor Niall , Sugrue Michael , Melly Conor , McGeehan Gearoid , Bucholc Magda , Crawford Aileen , O’Connor Paul , Abu-Zidan Fikri , Wani Imtiaz , Balogh Zsolt J. , Shelat Vishal G. , Tebala Giovanni D. , De Simone Belinda , Eid Hani O. , Chirica Mircea , Fraga Gustavo P. , Di Saverio Salomone , Picetti Edoardo , Bonavina Luigi , Ceresoli Marco , Fette Andreas , Sakakushe Boris , Pikoulis Emmanouil , Coimbra Raul , ten Broek Richard , Hecker Andreas , Leppäniemi Ari , Litvin Andrey , Stahel Philip , Tan Edward , Koike Kaoru , Catena Fausto , Pisano Michele , Coccolini Federico , Johnston Alison 
Published: March 2022

Abstract
Background
Despite the call to enhance accuracy and value of operation records few international recommended minimal standards for operative notes documentation have been described. This study undertook a systematic review of existing operative reporting systems for laparoscopic cholecystectomy (LC) to fashion a comprehensive, synoptic operative reporting template for the future.

Methods
A search for all relevant articles was conducted using PubMed version of Medline, Scopus and Web of Science databases in June 2021, for publications from January 1st 2011 to October 25th 2021, using the keywords: laparoscopic cholecystectomy AND operation notes OR operative notes OR proforma OR documentation OR report OR narrative OR audio-visual OR synoptic OR digital. Two reviewers (NOC, GMC) independently assessed each published study using a MINORS score of ≥ 16 for comparative and ≥ 10 for non-comparative for inclusion. This systematic review followed PRISMA guidelines and was registered with PROSPERO. Synoptic operative templates from published data were assimilated into one “ideal” laparoscopic operative report template following international input from the World Society of Emergency Surgery board.

Results
A total of 3567 articles were reviewed. Following MINORS grading 25 studies were selected spanning 14 countries and 4 continents. Twenty-two studies were prospective. A holistic overview of the operative procedure documentation was reported in 6/25 studies and a further 19 papers dealt with selective surgical aspects of LC. A unique synoptic LC operative reporting template was developed and translated into Chinese/Mandarin, French and Arabic.

Conclusion
This systematic review identified a paucity of publications dealing with operative reporting of LC. The proposed new template may be integrated digitally with hospitals’ medical systems and include additional narrative text and audio-visual data. The template may help define new OR (operating room) recording standards and impact on care for patients undergoing LC.
Perioperative incidence of ECMO and IABP on 5901 mitral valve surgery procedures
Authors: Condello Ignazio , Lorusso Roberto , Santarpino Giuseppe , Di Bari Nicola , Fiore Flavio , Moscarelli Marco , Calafiore Antonio Maria , Speziale Giuseppe , Nasso Giuseppe 
Published: March 2022

Abstract
Background
Report the incidence and results of peri-operative extracorporeal membrane oxygenation (ECMO) and intra-aortic balloon pump (IABP) of patients undergoing mitral valve surgery (MVS) through right mini-thoracotomy (RT) and conventional full sternotomy (FS) for a period of 6 years from eleven tertiary Cardiac Surgery Institutes of GVM Care & Research Italia.

Methods
From January 2016 to November 2021, a total of 5901 consecutive patients underwent MVS through RT and FS. The primary outcome of the study was the mortality and incidence of low cardiac output syndrome (LCOS) treated with intra-aortic balloon pump (IABP) with or without inotropic support and the incidence of Postcardiotomy Cardiogenic Shock (PCS) treated with Veno-arterial (VA) Extracorporeal Membrane Oxygenation (ECMO) on patients undergoing mitral valve surgery (MVS) through right mini-thoracotomy (RT) versus conventional full sternotomy (FS).

Results
The mean age was 66 ± 15 years, 3389 patients underwent in RT approach 2512 in FS, 3081 (52%) patients were male and 2.3% had previous cardiac operations. Cardiopulmonary bypass time was 93 min for RT and 81 min for FS and cross clamp time 75 min for RT and 63 min for FS for mitral valve repair. Incidence of perioperative IABP for the treatment of low cardiac output was reported on 99 patients (1.6%), 51 for RT (1.5%), 35% used inotropic support (adrenaline and milrinone) and 48 in FS (1.9), 28% use inotropic support, 21 patients died after IABP (3 RT and 18 FS). Incidence of perioperative VA-ECMO for the PCS treatment was 13 and 4 with IABP, 9 RT (0.2%) and 4 FS approach (0.15%), 12 patients died after VA-ECMO.

Conclusion
Minimally invasive mitral valve surgery is a safe and reproducible approach associated with low mortality and morbidity. ECMO and IABP incidence for the treatment of PCS was 0.2% and for Low cardiac output syndrome (LCOS) was 1.6% in elective mitral valve surgery is very low. The patients that use the perioperative IABP in minimally invasive mitral valve surgery (MIMVS) trough RT reported a reduced mortality compared to FS in relation to the operative risk and surgical technique. Low incidence of VA-ECMO was found in RT and FS approach, only one patient survived after VA-ECMO after minimally invasive mitral valve surgery.
BDNF and TrKB expression levels in patients with endometriosis and their associations with dysmenorrhoea
Authors: Wang Sha , Duan Hua , Li Bohan , Hong Wei , Li Xiao , Wang Yiyi , Guo Zheng Chen 
Published: March 2022

Abstract
Background
Brain-derived neurotrophic factor (BDNF) is a known regulator of the development and maintenance of chronic pain in various chronic disorders. Together with its high-affinity tyrosine kinase type B (TrKB) receptor, BDNF is extensively expressed in the mammalian female reproductive system. However, BDNF and TrKB expression in different stages of endometriosis and the relationship between the expression of each in ectopic lesions and endometriosis pain remain unclear.

Methods
Sixty-two women who underwent laparoscopic surgery were enrolled in this study: forty-six diagnosed with ovarian endometrioma (study group) and sixteen diagnosed with ovarian benign tumours (control group). Samples from eutopic endometrium and ovarian endometriotic lesions were obtained at laparoscopic surgery. BDNF and TrKB messenger RNA (mRNA) and proteins levels in the eutopic and ectopic endometrium of both groups were measured by real-time PCR and immunohistochemical staining, respectively. Before the surgery the visual analogue scale (VAS) was used to measure dysmenorrhoea.

Results
BDNF and TrKB expression levels were higher in ovarian endometriotic lesions than in eutopic endometrium and normal endometrium (P < 0.05), and there was no cyclical change. Furthermore, their expression levels were higher in eutopic endometrium than in normal endometrium (P < 0.05), and BDNF and TrKB levels were higher in stage IV ovarian endometriotic lesions than in stage II and III lesions (P < 0.05), with their expression being non-significantly higher in stage III than in stage II (P > 0.05). Additionally, correlation coefficients for the association analysis between the mRNA expression of BDNF or TrKB in eutopic endometrium and the dysmenorrhoea VAS score were r = 0.52 and r = 0.56 for BDNF and TrKB, respectively (P < 0.05). The correlation coefficients for the associations between BDNF and TrKB in both the eutopic and ectopic endometrium were r = 0.82 and r = 0.66, respectively (P < 0.05).

Conclusions
BDNF and TrKB are closely related to dysmenorrhoea caused by endometriosis and may be important in the pathobiology or pathophysiology of endometriosis.
CUT&Tag2for1: a modified method for simultaneous profiling of the accessible and silenced regulome in single cells
Authors: Janssens Derek H. , Otto Dominik J. , Meers Michael P. , Setty Manu , Ahmad Kami , Henikoff Steven 
Published: March 2022

AbstractCleavage Under Targets and Tagmentation (CUT&Tag) is an antibody-directed transposase tethering strategy for in situ chromatin profiling in small samples and single cells. We describe a modified CUT&Tag protocol using a mixture of an antibody to the initiation form of RNA polymerase II (Pol2 Serine-5 phosphate) and an antibody to repressive Polycomb domains (H3K27me3) followed by computational signal deconvolution to produce high-resolution maps of both the active and repressive regulomes in single cells. The ability to seamlessly map active promoters, enhancers, and repressive regulatory elements using a single workflow provides a complete regulome profiling strategy suitable for high-throughput single-cell platforms.
Erucin inhibits osteoclast formation via suppressing cell–cell fusion molecule DC-STAMP without influencing mineralization by osteoblasts
Authors: Takagi Tomohiro , Inoue Hirofumi , Fujii Shungo , Takahashi Nobuyuki , Uehara Mariko 
Published: March 2022

Abstract
Objective
Erucin (ERN), an isothiocyanate, is derived from the vegetable arugula. Although ERN has antitumor and antioxidant activity, the effect of ERN on osteoclast and osteoblast differentiation is not well documented. In this study, we evaluated the effects of ERN on osteoclast and osteoblast differentiation in vitro.

Results
ERN significantly reduced the formation of 1α,25(OH)2D3-induced tartrate-resistant acid phosphatase (TRAP)-positive cells at non-cytotoxic concentrations. Furthermore, ERN downregulated the mRNA expression of osteoclast-associated genes, such as nuclear factor of activated T cells cytoplasmic-1, TRAP, and cathepsin K. In addition, ERN suppressed mRNA expression of dendritic cell specific transmembrane protein (DC-STAMP), which encodes cell–cell fusion. However, ERN did not affect mineralization by osteoblasts. Thus, our data suggest that ERN may attenuate osteoclastic bone resorption by inhibiting multinucleation of mononuclear pre-osteoclasts and by suppressing mRNA expression of DC-STAMP in bone marrow cells without influencing mineralization by osteoblasts.
Comparison of circulating fibrocytes from non-asthmatic patients with seasonal allergic rhinitis between in and out of pollen season samples
Authors: Côté Marie-Ève , Boulay Marie-Ève , Plante Sophie , Côté Andréanne , Chakir Jamila , Boulet Louis-Philippe 
Published: March 2022

Abstract
Background
Allergic rhinitis is a risk factor for asthma development. In asthma, fibroblast progenitors, fibrocytes, are increased in the blood and bronchial mucosa following allergen exposure. These cells may play a role in lower airways remodeling as observed in non-asthmatic subjects with allergic rhinitis.

Objective
To determine the influence of seasonal allergen exposure on blood circulating fibrocytes in allergic rhinitic subjects without asthma.

Methods
Non-asthmatic subjects with seasonal allergic rhinitis had blood sampling at baseline and at the peak of rhinitis symptoms. Cells were stained for fibrocyte markers (CD34, CD45, CXCR4, collagen I) and analyzed by flow cytometry.

Results
Data from 26 subjects (11M:15F) aged 29 ± 8 years were analysed. Compared to baseline, there was a significant decrease in blood fibrocytes during the pollen season in subjects sensitized to trees [median (25–75 percentile), 9.3 (6.4–20.7)% vs 7.0 (4.2–10.1)%, P = 0.007] and a significant increase in subjects sensitized to grass [12.7 (9.9–23.1)% vs 64.0 (57.6–73.6)%, P < 0.001] and ragweed [8.0 (7.4–10.8)% vs 48.2 (43.5–52.6)%, P < 0.001]. A significant decrease in CXCR4 mean fluorescence was also observed between the two visits [1814 (1261–2235) vs 1352 (814–1796) (arbitrary units), P = 0.02].

Conclusions and clinical relevance
These results contribute to document dynamic variations in blood fibrocytes’ activation and migration into the airways following natural exposure to allergens. These findings may help identify one of the potential factors involved in the development of asthma in allergic rhinitic subjects.
Atypical presentation of an atypical pneumonia: a case report
Authors: Payus Alvin Oliver , Clarence Clarita , Nee Tiong , Yahya Wan Nur Nafisah Wan 
Published: March 2022

Abstract
Background
Neurologic impediments occur in only 0.1% of Mycoplasma pneumoniae infections. Although direct intracerebral infection can occur in these patients, autoimmune-mediated reactions secondary to molecular mimicry are the most common pathophysiology of such neurological complications. These complications include immune-mediated encephalitis, peripheral neuritis such as Guillain–Barré syndrome, and many others. Miller Fisher syndrome is a one of the variants of Guillain–Barré syndrome that has been rarely linked to Mycoplasma pneumoniae infection. It is a condition classically characterized by the triad of ophthalmoplegia, areflexia, and ataxia. Most patients with Miller Fisher syndrome will have positive anti-ganglioside GQ1b antibodies found in their serum, making this autoantibody a very useful serological confirmation parameter. We report a case of a Miller Fisher syndrome in a woman with Mycoplasma pneumoniae infection. To the best of the authors’ knowledge, such cases have been only rarely described in literature.

Case presentation
A 35-year-old Chinese woman presented with sudden onset of double vision and ataxia 5 days after fever and mild flu symptoms. Her Mycoplasma pneumoniae antigen was positive with 1 over 2560 titer of total mycoplasma antibody and presence of immunoglobulin M antibody, suggesting acute infection, and her nerve conduction study revealed mild sensory axonal polyneuropathy with segmental demyelination. the Miller Fischer syndrome variant of Guillain-Barré syndrome secondary to Mycoplasma pneumonia was suspected and later confirmed by presence of serum anti-GQ1b autoantibody. She was treated with intravenous immunoglobulin 0.4 g/kg once daily for 5 days.

Conclusions
The objective of this report is to share a case of an uncommon neurological complication of Mycoplasma pneumoniae infection, to increase the level of suspicion among clinicians that Miller Fischer syndrome can occur as an atypical presentation of an atypical pneumonia.
Half-linear differential equations of fourth order: oscillation criteria of solutions
Authors: Bazighifan Omar , Al-Ghafri Khalil S. , Al-Kandari Maryam , Ghanim F. , Mofarreh Fatemah 
Published: March 2022

AbstractIn this paper, we are concerned with the oscillation of solutions to a class of fourth-order delay differential equations withp-Laplacian like operators$( r ( t ) \vert x^{\prime \prime \prime } ( t ) \vert ^{p_{1}-2}x^{\prime \prime \prime } ( t ) ) ^{\prime }+q ( t ) \vert x ( \tau ( t ) ) \vert ^{p_{2}-2}x ( \tau ( t ) ) =0$(r(t)|x‴(t)|p1−2x‴(t))′+q(t)|x(τ(t))|p2−2x(τ(t))=0and$( r ( t ) \vert x^{\prime \prime \prime } ( t ) \vert ^{p_{1}-2}x^{\prime \prime \prime } ( t ) ) ^{\prime }+\sigma ( t ) \vert x^{\prime \prime \prime } ( t ) \vert ^{p_{1}-2}x^{ \prime \prime \prime } ( t ) +q ( t ) \vert x ( \tau ( t ) ) \vert ^{p_{2}-2}x ( \tau ( t ) ) =0$(r(t)|x‴(t)|p1−2x‴(t))′+σ(t)|x‴(t)|p1−2x‴(t)+q(t)|x(τ(t))|p2−2x(τ(t))=0. New oscillation criteria are presented by the comparison technique and employing the Riccati transformation. Moreover, our results are an extension and complement to previous results in the literature. Two examples are shown to illustrate the conclusions.
Environmental surveillance of ESBL and carbapenemase-producing gram-negative bacteria in a Ghanaian Tertiary Hospital
Authors: Acolatse Joseph Elikem Efui , Portal Edward A. R. , Boostrom Ian , Akafity George , Dakroah Mavis Puopelle , Chalker Victoria J. , Sands Kirsty , Spiller Owen B. 
Published: March 2022

Abstract
Background
The burden of antibiotic resistant infection is mainly felt in low-to-middle income countries, where the rate of antimicrobial resistance is largely under-surveyed and under huge pressure from unregulated, disparate and often self-guided access to antimicrobials. Nosocomial infections from hospital environments have been shown to be a particularly prevalent source of multi-drug resistant strains, yet surveillance of hospital environmental contamination is often not investigated.

Methods
The study was prospective, observational and cross-sectional, sampling 231 high and low touch surfaces from 15th March to 13th April 2021, from five wards in the Cape Coast Teaching Hospital, Ghana. Microbial growth in the presence of vancomycin and either meropenem or cefotaxime was examined and bacterial species were identified by MALDI-TOF. The presence of common extended-spectrum β-lactamases (ESBL) and carbapenemase antimicrobial resistance genes (ARG) were identified through PCR screening, which were confirmed by phenotypic antimicrobial susceptibility determination. Isolates positive for carbapenem resistance genes were sequenced using a multi-platform approach.

Results
We recovered microbial growth from 99% of swabs (n = 229/231) plated on agar in the absence of antimicrobials. Multiple sites were found to be colonised with resistant bacteria throughout the hospital setting. Bacteria with multi-drug resistance and ARG of concern were isolated from high and low touch points with evidence of strain dissemination throughout the environment. A total of 21 differing species of bacteria carrying ARG were isolated. The high prevalence of Acinetobacter baumannii carrying blaNDM-1 observed was further characterised by whole genome sequencing and phylogenetic analysis to determine the relationship between resistant strains found in different wards.

Conclusion
Evidence of multiple clonal incursions of MDR bacteria of high sepsis risk were found in two separate wards for a regional hospital in Ghana. The prevalence of multiple blaNDM carrying species in combination with combinations of ESBLs was particularly concerning and unexpected in Africa. We also identify strains carrying tet(X3), blaVIM-5 or blaDIM-1 showing a high diversity of carbapenamases present as a reservoir in a hospital setting. Findings of multi-drug resistant bacteria from multiple environmental sites throughout the hospital will inform future IPC practices and aid research prioritisation for AMR in Ghana.
Implementation of an e-Learning course in physical activity and sedentary behavior for pre- and in-service early childhood educators: Evaluation of the TEACH pilot study
Authors: Bruijns Brianne A. , Vanderloo Leigh M. , Johnson Andrew M. , Adamo Kristi B. , Burke Shauna M. , Carson Valerie , Heydon Rachel , Irwin Jennifer D. , Naylor Patti-Jean , Timmons Brian W. , Tucker Patricia 
Published: March 2022

Abstract
Background
Childcare-based physical activity (PA) and sedentary behavior (SB) interventions have traditionally used in-person training to supplement early childhood educators’ (ECEs) knowledge and confidence to facilitate physically active programming for the children in their care. However, this method of delivery is resource-intensive and unable to reach a high number of ECEs. The purpose of the Training pre-service EArly CHildhood educators in PA (TEACH) pilot study was to test the implementation (e.g., fidelity, feasibility, acceptability) of an e-Learning course targeting PA and SB among a sample of pre-service (i.e., post-secondary students) and in-service (i.e., practicing) ECEs in Canada.

Methods
A pre-/post-study design was adopted for this pilot study, and implementation outcomes were assessed cross-sectionally at post-intervention. Pre-service ECEs were purposefully recruited from three Canadian colleges and in-service ECEs were recruited via social media. Upon completing the e-Learning course, process evaluation surveys (n = 32 pre-service and 121 in-service ECEs) and interviews (n = 3 pre-service and 8 in-service ECEs) were completed to gather ECEs’ perspectives on the e-Learning course. Fidelity was measured via e-Learning course metrics retrieved from the web platform. Descriptive statistics were calculated for quantitative data, and thematic analysis was conducted to analyze qualitative data.

Results
Moderate-to-high fidelity to the TEACH study e-Learning course was exhibited by pre-service (68%) and in-service (63%) ECEs. Participants reported that the course was highly acceptable, compatible, effective, feasible, and appropriate in complexity; however, some ECEs experienced technical difficulties with the e-Learning platform and noted a longer than anticipated course duration. The most enjoyed content for pre- and in-service ECEs focused on outdoor play (87.5% and 91.7%, respectively) and risky play (84.4% and 88.4%, respectively).

Conclusions
These findings demonstrate the value of e-Learning for professional development interventions for ECEs. Participant feedback will be used to make improvements to the TEACH e-Learning course to improve scalability of this training.