bluuebunny/crossref_metadata_2025_split huggingface dataset:
Efficient in vitro and in vivo self-repression of SpCas9 gene using a molecular Hara-Kiri method.
Authors: Iyombe-Engembe Jean-Paul , Duchene Benjamin , Rousseau Joel , Ouellet Dominique , Cherif Khadija , Guyon Antoine , Tremblay Jacques P. 
Published: March 2022

Abstract
The CRISPR/Cas9 system is currently a major revolution in the field of biology. Because of its simplicity compared to other endonucleases, this system is being experimented in diverse fields. However, a major disadvantage is the toxicity linked to sustained Cas9 expression. In the present study, we present an approach to effectively suppress the expression of the Streptococcus pyogenes Cas9 (SpCas9) gene. This approach that we call the molecular Hara-Kiri method, involves two sgRNAs targeting two sequences in the SpCas9 gene. The SpCas9 enzyme binds to the Protospacer Adjacent Motifs following the two sequences targeted by the sgRNAs and induces two Double Strand Breaks (DSBs) in its own gene (Hara-Kiri). The sequence located between the DSBs is then deleted. Most of the time, the SpCas9 gene is repaired by Non-Homologous End Joining without INDELs. By adequately selecting the targeted sequences, the junction of the SpCas9 gene residues generates a TAA type stop codon within this truncated gene to effectively suppress its expression. This results in dramatic decrease of the SpCas9 protein in vitro and in vivo.
Forest heterogeneity outweighs movement costs by enhancing hunting success and fitness in spotted owls
Authors: Zulla Ceeanna J , Jones Gavin M. , Kramer H. Anu , Keane John J. , Roberts Kevin N. , Dotters Brian P. , Sawyer Sarah C. , Whitmore Sheila A. , Berigan William J. , Kelly Kevin G. , Gutiérrez R. J. , Peery M. Zachariah 
Published: March 2022

Abstract
ContextThe concepts of habitat fragmentation and heterogeneity are central to the conservation of biodiversity; yet understanding when landscapes transition from heterogenous to fragmented can challenge conservation in practice. Complex and sometimes difficult-to-measure responses of species, and ultimately biological communities, to habitat heterogeneity and fragmentation may reflect the outcome of life-history trade-offs shaped by different landscape properties. ObjectivesHere, we tested the hypothesis that a mosaic of forest stands improved hunting and breeding success for California spotted owls (Strix occidentailis occidentalis). MethodsWe integrated high-temporal-resolution GPS tags, video monitoring of nests sites, long-term assessments of reproductive status, and high-resolution remotely-sensed vegetation data in a mixed-ownership landscape in the Sierra Nevada, California. ResultsSpotted owls made shorter nocturnal movements when medium forest was prevalent in their territory. However, spotted owls delivered prey at a higher rate to nests sites when they had more forest edge in their territory, which presumably provided greater access to large-bodied woodrat prey. Further, spotted owl reproductive output was relatively high in territories that contained a mix of mature and open forest. ConclusionsThus, the benefits heterogenous forests provide to hunting success appear to outweigh costs associated with additional commuting to foraging sites and provide emergent fitness benefits to spotted owls. We suggest that the line between habitat heterogeneity and fragmentation can be a complex one that varies not only among, but within, species. Further, understanding the effects of heterogeneity and fragmentation on biological communities will require more empirical and mechanistic studies of individual species.
The Impact of 2009 Versus 2021 Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) GFR Estimating Equations on CKD Prevalence in Asians
Authors: Betzler Bjorn , Sultana Rehena , Feng He , Tham Yih Chung , Lim Cynthia Ciwei , Wang Ya Xing , Nangia Vinay , Tai E Shyong , Rim Tyler Hyungtaek , Bikbov Mukharram M , Jonas Jost B , Kang Se Woong , Park Kyu Hyung , Cheng Ching-Yu , Sabanayagam Charumathi 
Published: March 2022

Abstract
Background: In 2021, the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) validated a new equation for estimated glomerular filtration rate (eGFR) without race as an explanatory variable. Because the prevalence of chronic kidney disease (CKD) in Asians is known to differ from other ethnicities, this manuscript compares prevalence of CKD in multiple Asian cohorts using the 2009 and 2021 creatinine-based eGFR equations.Methods: Eight population-based studies from China, India, Russia (Asian), Singapore and South Korea provided individual-level data (n=67,233). CKD was defined as an estimated glomerular filtration rate (eGFR) <60 mL/min/1.73m2 (G3-G5). GFR was estimated using both the 2009 CKD-EPI equation developed using creatinine, age, sex, and race (eGFRcr [2009, ASR]) and the 2021 CKD-EPI equation developed without race (eGFRcr [2021, AS]). Prevalence of eGFR categories was compared within each study and within subgroups of age, gender, body mass index (BMI), diabetes, and hypertension status. The extent of reclassification was examined using net reclassification improvement (NRI). Results: Of 67,233 adults, CKD prevalence was 8.6% (n=5800/67,233) using eGFRcr (2009, ASR) and 6.4% (n=4307/67,233) using eGFRcr (2021, AS). Within individual studies, all 8 studies saw a reduction in CKD prevalence, ranging from -7.0% (95% CI -8.5% to -5.4%) to -0.4% (-1.3% to 0.5%). A reduction in CKD prevalence when using eGFRcr (2021, AS) was observed in all subgroups, except those in the BMI < 18.5% subgroup. Net reclassification index (NRI) was significant at -2.33% (p<0.001). No individuals were reclassified as a higher (more severe) eGFR category, while 1.7-4.2% of individuals with CKD were reclassified as one eGFR category lower when eGFRcr (2021, AS) rather than eGFRcr (2009, ASR) was used.Conclusions: eGFRcr (2021, AS) consistently provided reduced CKD prevalence and GFR estimates among Asian cohorts than eGFRcr (2009, ASR). Comparison of both equations to predict progression to renal failure or adverse outcomes using prospective studies are warranted for accurate validation within Asian populations.
Medical Travel For Surgical Procedures: A Systematic Review Of The Literature
Authors: Weinberg Fabienne Ingrid , Wagstaff Duncan , Moonesinghe Ramani , Vindrola-Padros Cecilia 
Published: March 2022

Abstract
Background: Thousands of patients travel each year to access surgical procedures in other locations but there is limited evidence on the types of procedures they seek, the coordination of care across countries, the role played by their companion-caregivers and the presence of complications. Aim: The current review aims to provide an overview of the current evidence on medical travel for surgical procedures.Methods: A systematic review of peer-reviewed articles was conducted by searching the databases PubMed, Web of Science, ProQuest Central and CINAHL for papers on medical travel for surgeries. Data were extracted through a form developed in REDCap. The MMAT was used to assess the quality of the studies. We followed PRISMA. Results: The review included a total of 58 articles. Patients travelled with their caregiver-companions to gain access to surgical procedures. Pre-assessments in the destination countries were limited. Access to follow-up care was challenging, threatening the continuity of care. There was limited information exchange between sending and destination countries. Significant health risks for patients were identified in the form of postoperative complications.Conclusion: Future research needs to explore the full pathway of care of patients and their caregiver-companions traveling to access surgical procedures. Protocol registration: CRD: 42018114495
Spatial dynamics of urbanization and refugees’ settlements: Case study evidence using mixed method
Authors: Islam Md Tariqul , Charlesworth Mark , Rabbi Atta , Sikder Sujit Kumar 
Published: March 2022

Abstract
Urbanization needs to conceptualize beyond the city boundaries by exploring complex and interconnected interdependencies in regional key driving factors that can have tele-coupling effects during sudden shocks. Massive refugee settlement has a robust inter-linkage to the spatial and socio-ecological dynamics in local, regional contexts in both global south and North Countries. This study aims to explore the spatial dimension of urbanization in a regional transition context focusing on the newly built refugee settlement in the south-eastern coastal area of Bangladesh. For locational proximity and comparatively easy escape from Myanmar after domestic conflicts, over one million Rohingya have settled in the rural/forest area in Cox’s Bazar. Analysis of openly available global data products from historical built-up expansion from 1975 to 2021 suggests that refugee settlements impact spatial development intensity and population density dimensions, which have a transition (rural-to-urban) of a greater regional scale than local urbanization. Although our results from the expert-based key informant interviews have a broad agreement with the quantitative findings; however, the environmental, socio-economic, and cultural impacts are more local than findings from the geospatial analysis. Therefore, the local development policies need an urgent adjustment to comply with the local and regional balances in the above factors, along with international development actors may also need attentive measures in the policy formulation. Further investigations in scenario-based urbanization dynamics are required to avoid an urban desert initiated in the absence or relocation of refugee settlement. Similar kinds of studies may also be replicated even in prosperous global north countries in order to conceptualize the spatial transformative process of rapid migration influx on urbanization with innovative open data in the absence of up-to-date official datasets and adoptions of mixed approaches including related actors and policymakers.
Clinical significance of p53 protein expression and TP53 mutation status in colorectal cancer
Authors: Kim Kyoung Min , Ahn Ae-Ri , Park Ho Sung , Jang Kyu Yun , Moon Woo Sung , Kang Myoung Jae , Ha Gi Won , Lee Min Ro , Chung Myoung Ja 
Published: March 2022

Abstract
In human colorectal cancer (CRC), TP53 is one of the most important driver genes. Immunohistochemistry (IHC) has been used most often to assess the mutational status of TP53. Recently, next-generation sequencing (NGS) of the TP53 gene has increased. However, to our knowledge, a comparison between TP53 status evaluated by IHC and NGS has not been studied. Therefore, the primary aim of this study was to compare the clinical effect of TP53 status evaluated by IHC and NGS in patients with CRC. The secondary aim was to investigate the correlation between expression of p53 by IHC and mutational status of TP53 by NGS. We performed immunohistochemical staining of p53 and sequencing of TP53 by NGS in 204 human samples of CRC. We then analyzed the correlation between mutational status of TP53 and p53 expression, along with their prognostic impact in CRC patients. There was significant correlation between p53 expression and TP53 mutation (P < 0.001), TP53 mutation and higher N stage (P = 0.024), and positive p53 expression and higher N stage (P = 0.003). Positive p53 expression (P = 0.018) was significantly associated with overall survival (OS) of CRC patients by univariate analysis and was revealed as an independent prognostic factor by multivariate analysis. Additionally, the nonsense/frameshift (p53 proportion, 0%) p53 expression pattern showed a significantly better prognosis than the wild type and missense p53 expression patterns. However, the mutational status of TP53 was not significant in OS of CRC patients. These results suggest that IHC expression of p53 protein correlates with mutation status of TP53 and expression of p53 protein rather than mutation status of TP53 has more significant impact on the OS of CRC patients.
Cervicovaginal microflora isolated from healthy women exhibited probiotic properties and antimicrobial activity against pathogens isolated from cervical cancer patients
Authors: Kamble Ashwini , Naik Shilpa , Talathi Manju , Jadhav Deepali , Pingale Samradni , Kaul-Ghanekar Ruchika 
Published: March 2022

Abstract
Abnormal cervicovaginal microflora plays an important role in HPV persistence and progression to cervical cancer. The present study aimed at isolating and identifying probiotics from vaginal swabs of healthy women and evaluating their activity against vaginal pathogens isolated from cervical cancer patients. Based on probiotic, acid-bile tolerance and antimicrobial properties, 13 lactic acid bacteria (LAB) from the healthy group were identified by MALDI TOF MS. Among these, four strains, Lactobacillus gasseri P36Mops, Limosilactobacillus fermentum P37Mws, Lactobacillus delbrueckii P31Mcs and Enterococcus faecium P26Mcm, exhibited significant antimicrobial activity against 8 vaginal pathogens (Staphylococcus haemolyticus P41Tcs, Escherichia coli P30Tcs, E. coli P79Bcm, Enterococus faecalis P29Mops, E. faecalis P50Tws, E. faecalis P68Tcb, S. haemolyticus P48Bcb and S. haemolyticus P58Bcb) isolated from precancerous and cervical cancer patients. 16S rRNA sequencing of four potential probiotics revealed congruency with the MALDI-TOF MS (Matrix Assisted Laser Desorption and Ionisation, Time Of Flight Mass Spectrometry) identification and phylogenetic analysis showed genetic relationship with previously reported LAB strains. The selected LAB showed strain specific hydrophobicity (35.88-56.70%), auto-aggregation (35.26-61.39%) and antibiotic susceptibility. Interestingly, L. gasseri P36Mops was resistant to five standard antibiotics routinely used against urogenital or vaginal infections. LCMS (Liquid Chromatography Mass Spectrometry) analyses of the CFS (cell-free supernatant) of the four probiotics revealed presence of metabolites such as N-(1-deoxy-1-fructosyl)valine, hygroline, acetoxy-2-hydroxy-16-heptadecen-4-one, avocadyne 4-acetate, avocadyne 2-acetate, taraxinic acid glucosyl ester, 6-hydroxypentadecanedioic acid, with reported antimicrobial activity. The overall data suggests the bio-therapeutic potential of the identified vaginal probiotics against cervical cancer associated pathogens.
Value of Indocyanine Green Tracing Combined with Ex-Vitro Anatomical Sorting of Lymph Nodes in Clinical Prognosis of Patients with Gastric Cancer
Authors: Li Jieming , Liu Dongning , Jiang Qunguang 
Published: March 2022

Abstract
Objective: In order to increase the detection rate of lymph nodes (LNs) in radical gastrostomy, we applied indocyanine green (ICG) tracing and Ex vitro anatomical sorting to LNs dissection, and explored the guiding role of LNs number detected in the prognosis of patients with gastric cancer (GC). Methods: The clinical data of 88 GC patients who underwent D2 radical gastrostomy were retrospectively analyzed, and they were divided into ICG group and non-ICG group according to ICG usage and LN sorting method. Total LN number, positive LN number, clinical treatment, tumor marker, and patients’ prognosis between two groups were compared. Furthermore, we discussed the correlation between total LN and positive LN, and evaluated the role of ICG tracing in their detection. Results: Total LNs number, total positive LNs number, and LNs number in different LN sub stations detected in ICG group were higher. Furthermore, positive LNs number was positively correlated with total LNs number. In the evaluation of ICG tracing effect on LNs, it was found that fluorescent labeled LNs accounted for nearly 62% of total LNs and about 74% of positive LNs. In terms of clinical prognosis, the levels of CEA, CA199, CA242, and CA724 in ICG group were better controlled. Besides, GC patients’ in ICG group were not prone to relapse and metastasize, and their long-term survival rate were better. Conclusion: ICG tracing combined with Ex vitro anatomical sorting can significantly improve the detection rate of total LNs and positive LNs, which has a positive impact on patients’ long-term prognosis.
From Microscopic To Endoscopic Transsphenoidal Surgery: A Single Institution’s Learning Curve In Pituitary Tumor Treatment
Authors: Aman Renindra Ananda , Perkasa Sayyid Abdil Hakam , Sitorus Risayogi Wicaksana , Anindya Yodie , Wicaksana Bima Andyan , Kurniawan Ramadhan 
Published: March 2022

Abstract
Introduction: Transsphenoidal surgery with an operative microscope was a standard procedure for pituitary adenomas resection in our institution. Still, since the end of 2020, we have shifted the protocol to endoscopic transsphenoidal approach. This paper presented our learning curve in pituitary surgery using endoscopy instruments.Method: Patients with pituitary tumors who received transsphenoidal surgery during 2019 – 2021 were included. We compared the extent of resection, re-operation rate, blood loss volume, operative time, and complications of postoperative cerebrospinal fluid (CSF) leakage and diabetes insipidus (DI) between microscopic and endoscopic transsphenoidal approaches. Tumors extension to the suprasellar area and cavernous sinus were also analyzed to see their relationship with surgical outcomes.Result: Twenty-seven (55.1%) and 22 (44.9%) patients underwent microscopic transsphenoidal and endoscopic transsphenoidal surgery. There was no significant difference in total removal rate, re-operation rate, post-operative CSF leakage rate, blood loss volume, and operative time between the two approaches. Patients who received endoscopic surgery had a higher rate of DI (P = 0.002). Tumors extension to cavernous sinus was a significant predictor of total resection rate (P = 0.002). Multivariate analysis showed that surgical approach, suprasellar and cavernous sinus extension did not significantly affect the extent of surgery.Conclusion: Our endoscopic and microscopic surgery results are comparable despite the limitation of experience with endoscopy instruments. This report supports our transition from microscopic to endoscopic pituitary surgery.
Harpagoside Attenuates Local Bone Erosion and Systemic Osteoporosis in Collagen-Induced Arthritis in Mice
Authors: Kim Ju-Young , Cheon Yoon-Hee , Ahn Sung-Jun , Kwak Sung Chul , Chung Chong Hyuk , Lee Chang Hoon , Lee Myeung Su 
Published: March 2022

Abstract
Background: Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease that causes local bone erosion and systemic osteoporosis. Harpagoside (HAR), an iridoid glycoside, has various pharmacological effects on pain, arthritis, and inflammation. Our previous study suggests that HAR is more deeply involved in the mechanism of bone loss caused by inflammatory stimuli than hormonal changes. Here, we identified the local and systemic bone loss inhibitory effects of HAR on RA and its intracellular mechanisms using a type 2 collagen-induced arthritis (CIA) mouse model. Methods: The anti-osteoporosis and anti-arthritic effects of HAR were evaluated on bone marrow macrophage in vitro and CIA in mice in vivo by obtaining clinical scores, measuring hind paw thickness and inflammatory cytokine levels, micro-CT and histopathological assessments, and cell-based assay.Results: HAR markedly reduced the clinical score and incidence rate of CIA in both the prevention and therapy groups. Histological analysis demonstrated that HAR locally ameliorated the destruction of bone and cartilage and the formation of pannus. In this process, HAR decreased the expression of inflammatory cytokines, such as tumor necrosis factor-a, interleukin (IL)-6, and IL-1b in the serum of CIA mice. Additionally, HAR downregulated the expression of receptor activator of nuclear factor-kB ligand and upregulated that of osteoprotegerin. HAR suppressed systemic bone loss by inhibiting osteoclast differentiation and osteoclast marker gene expression in a CIA mouse model. Conclusions: Taken together, these findings show the beneficial effect of HAR on local symptoms and systemic bone erosion triggered by inflammatory arthritis.
Syringin Exerts Anti-Breast Cancer Effects Through PI3K-AKT and EGFR-RAS-RAF Pathways
Authors: Wang Fei , Yuan Chong , Liu Bo , Yang Fang Yan , Wu Zhen He 
Published: March 2022

Abstract
Background
Breast cancer (BC) is one of the most common malignant tumors with the highest mortality in the world. Modern pharmacological studies have shown that Syringin has an inhibitory effect on many tumors, but its anti-BC efficacy and mechanism are still unclear.
Methods
First, Syringin was isolated from Acanthopanax senticosus (Rupr. & Maxim.) Harms (AS) by systematic solvent extraction and silica gel chromatography column. Then, the potential mechanism of syringin against BC was revealed by bioinformatics and in vitro experiments. To provide a more experimental basis for later research, the hub genes which could be candidate biomarkers of BC and a ceRNA network related to them were obtained.
Results
Syringin was obtained by liquid chromatography-mass spectrometry (LC-MS), nuclear magnetic resonance (NMR), and high-performance liquid chromatography (HPLC). Bioinformatics results showed that MAP2K1, PIK3CA, HRAS, EGFR, Caspase3, and PTGS2 were the hub targets of Syringin against BC. In vitro experiments confirmed that Syringin inhibited the proliferation and migration and promoted apoptosis of BC cells through the above hub targets. And PIK3CA and HRAS were related to the survival and prognosis of BC patients, the PIK3CA-has-mir-139-5p-LINC01278 and PIK3CA-has -mir-375 pathways might be closely related to the mechanism of Syringin against BC.
Conclusions
Syringin against BC via PI3K-AKT-PTGS2 and EGFR-RAS-RAF-MEK-ERK pathways, and PIK3CA and HRAS are hub genes for adjuvant treatment of BC.
The Primary Care COVID-19 Integrated Pathway: A Rapid Response to Health and Social Impacts of COVID-19
Authors: Aghajafari Fariba , Hansen Brian , McBrien Kerry , Leslie Myles , Chiew Alexandra , Ward Rick , Li Bing , Hu Jia 
Published: March 2022

Abstract
Background
The first wave of COVID-19 in Calgary, Alberta accelerated the integration of primary care with the province’s centrally managed health system. This integration aimed to deliver wraparound in-community patient care through two interventions that combined to create the COVID-19 Integrated Pathway (CIP). The CIP’s interventions were: 1) a data sharing platform that ensured COVID-19 test results were directly available to family physicians (FPs), and 2) a clinical algorithm that supported FPs in delivering in-community follow up to improve patient outcomes. We evaluate the CIP by describing its function and capacity to facilitate FP follow-up with COVID-19 patients.
Method
We generated descriptive statistics by analyzing data from a Calgary Zone hub clinic called the Calgary COVID-19 Care Clinic (C4), provincially maintained records of hospitalization, Emergency Department (ED) visits, and physician claims.
Results
Between Apr. 16 and Sep. 27, 2020, 7289 patients were referred by the Calgary Public Health team to the C4 clinic. Of those, 48.6% were female, the median age was 37.4 y. 97% of patients had at least one visit with a healthcare professional, where follow-up was conducted using the CIP’s algorithm. 5.1% of patients visited an ED and 1.9% were hospitalized within 30 days of diagnosis. 75% of patients had a median of 4 visits with their FP.
Discussion
Our data suggest that that information exchange between PC and central systems facilitates primary care-based management of patients with COVID-19 in the community and has potential to reduce acute care visit.