bluuebunny/crossref_metadata_2025_split huggingface dataset:
Optimized Calcium-Phosphate based co-transfection of TAU and tdTomato into human iPSC-derived neurons for the study of intracellular distribution of wildtype and mutant human TAU.
Authors: Lilis Panagiotis , Kabbani Mohamed Aghyad Al , Zempel Hans 
Published: June 2023

Abstract
The study of TAU protein in disease-relevant neuronal cells in culture requires efficient delivery systems for transfection of exogenous TAU and also modulators and interactions of TAU. Transfection of cultivated cells using calcium phosphate precipitation is a simple and cost-effective approach, also for difficult-to-transfect and sensitive cells such as primary neurons. Because of its low cell toxicity and ease of use, the Ca2+-phosphate transfection method is one of the most widely used gene transfer procedures in neuroscience. However, Ca2+-phosphate transfection efficacy in neurons is poor, often in the range of 1–5%, limiting its use in functional investigations. Here, we outline our improved Ca2+-phosphate transfection methodology for human iPSC-derived neurons that yields a reasonable efficiency (20-30% for bright volume markers) without apparent effects on cell health. We have used it to introduce wildtype and mutant human TAU with and without co-transfection of a volume marker (used here: tdTomato). In sum, our procedure can deliver neuronal genes (e.g. MAPT) using typical eukaryotic expression vectors (e.g. using CMV promoter), and is optimized for transfection of human iPSC-derived neurons.
Sequential Glycosylations at the Multibasic Cleavage Site of SARS-CoV-2 Spike Protein Regulate Viral Activation, Assembly, and Infection
Authors: Wang Shengjun , Ran Wei , Sun Lingyu , Fan Qingchi , Zhao Yuanqi , Wang Bowen , Yang Jinghong , He Yuqi , Wu Ying , Wang Yuanyuan , Chen Luoyi , Chuchuay Arpaporn , Zhu Xinhai , Wang Yanqun , chen Yao-Qing , Yuan Yanqiu , Zhao Jincun , Mao Yang 
Published: June 2023

Abstract

The multibasic furin cleavage site at the S1/S2 boundary of the spike protein (S protein) is a hallmark of SARS-CoV-2 and is essential for its increased infectivity. O-glycosylation near the furin site catalyzed by host cell glycosyltransferases can theoretically hinder spike protein processing and impede viral infection, but so far such hypothesis has not been tested with authentic viruses. The mechanism for furin activation is not clearly understood either. Here in this study, we discovered that GalNAc-T3 and T7 together initiate clustered O-glycosylations in the multibasic S1/S2 boundary region, which inhibits furin processing of the spike protein and surprisingly suppresses the incorporation of S protein into virus-like-particles (VLPs). Mechanistic analysis revealed that the assembly of spike protein into VLPs relies on protein-protein interaction between the furin-cleaved S protein and a double aspartic motif on the membrane protein of SARS-CoV-2, suggesting a novel mechanism for furin activation of S protein. Interestingly, a point mutation at P681, found in the SARS-CoV-2 variants alpha and delta, resists the glycosylation by GalNAc-T3 and T7 and its inhibitory effect against furin processing. However, an additional mutation at N679 in the most recent omicron variant reverts this resistance, making it both prone to glycosylation in vitro and sensitive to the expression of GalNAc-T3 and T7 in human lung cells. Together, our results suggest a glycosylation-based defense mechanism of host cells against SARS-CoV-2 and reveal the host-pathogen interplay at this critical “battle field” as the virus first escapes and currently surrenders itself to the host cell glycosylation.
Dose-effect relationship of metaraminol before spinal anesthesia in preventing hypotension induced by spinal anesthesia in parturients undergoing cesarean section
Authors: Xue Xiaoli , Wang Jing , Zhu Chuling , Chen Xiaofen , Ma Simeng , Lin Zhoujin , Zhang Longxin , Zhou Min , Wei Ying 
Published: June 2023

Abstract
Objective:
To study the dose-effect relationship of metaraminol in preventing hypotension induced by spinal anesthesia in parturients undergoing cesarean section.
Methods:
One hundred and twenty-five parturients undergoing elective cesarean sections were randomly divided into 5 groups (n=25 each), metaraminol groups (PM1, PM2, PM3, PM4 and PM5). Metaraminol 20, 30, 40, 50 and 60μg·min-1 were intravenously infused before spinal anesthesia.
Effective prevention of hypotension was defined as the absence of hypotension during the period from the end of the administration of spinal anesthesia to the delivery of the fetus. The ED50, ED90, and 95% confidence intervals of metaraminol in preventing hypotension induced by spinal anesthesia were calculated by Probit analysis.
Results:
The ED50 and 95% confidence interval of metaraminol in preventing hypotension induced by spinal anesthesia were 46.52 (39.41-57.64) μg·min-1, and ED90 and 95% confidence interval were 97.10(77.17-157.49) μg·min-1. The incidence of hypotension and shivering decreased with the increase in the dose of metaraminol infused in the five groups (P<0.05). The incidence of reactive hypertension increased with the increase in the dose of metaraminol infused in the five groups (P<0.05).
Conclusion:
The ED50 and ED90 of metaraminol before spinal anesthesia in preventing spinal anesthesia-induced hypotension, in the patients undergoing cesarean section are 46.52 and 97.10μg·min-1, respectively.
In silico and in vitro study of the anti-tuberculous activity of Nigella sativa L. bioactive compounds
Authors: Khamjan Nizar A. , Tabassum Heena , Khan Elhan , Lohani Mohtashim , Haque Mazharul , Mohammed Bahja Siddig , Maguen Elsie V. , Farasani Abdullah , Ahmad Iffat Zareen 
Published: June 2023

Abstract
Tuberculosis (TB) is a deadly disease which killed 1.6 million people in the year 2021 as per the reports of WHO. The treatment of the disease becomes more difficult once drug resistant TB strains (MDR and XDR TB) develops and its co-infection with HIV AIDS. There is a serious need to investigate and develop novel, cost-effective and potent anti-TB drugs. There is a requirement for the evaluation of the herbal drugs by standard scientific methods so as to validate them for the treatment of diseases. In the quest for the effective treatment, the methanolic extract of N. sativa L. sprouts was studied for its efficacy against M. tuberculosis (H37Ra) by Middlebrook broth assay. It was highly active and inhibited the growth at a low concentration of 100µ g/ml upto day 14th in comparison to the control along with the positive fluorescence test of the strain. Furthermore, both compounds showed high drug likeliness and ADME characteristics with no toxicity for oral administration in in silico studies. Molecular docking and dynamics studies also showed that thymoquinone and thymol have a significant binding potential with stable binding interaction to inhibit the Mycobacterium protein function, affirming it as a potent ant-TB drug.
Explanted skull flaps after decompressive hemicraniectomy demonstrate relevant bone avitality. Is their reimplantation worth of the risk?
Authors: Gousias Konstantinos , Stricker Ingo , Hoyer Annika , Theocharous Theocharis , Rompf Csilla , Pranata Arthur , Tannapfel Andrea , Agrawal Rachit , Tischoff Iris 
Published: June 2023

Abstract
Objective Reimplantations of autologous skull flaps after decompressive hemicraniectomies (DH) are associated with dramatic high rates of postoperative bone flap resorption (BFR). In the context of our clinical trial DRKS00023283, we assessed histologically the cell viability of explanted bone flaps in certain periods of time after DH, in order to conclude whether precursors of BRF may be developed during their storage.Methods Skull bone flaps explanted during a DH between 2019 and 2020 for a vascular disease or traumatic brain injury were sterile stored in a freezer at either − 23°C or -80°C. After their thawing process, the skulls have been collected for histological investigations. Parameters of bone metabolism, namely PTH1 and OPG have been analyzed via immunohistochemistry. H&E stain was used to assess the degree of avital bone tissue, whereas the repeated assays were performed after 6 months.Results A total of 17 stored skull flaps (8–23°C; 9–80°C) were analyzed. Median age of our cohort was 70 years; 9 patients (53%) were male. Duration of cryopreservation varied between 2 and 17 months. Relevant degree of bone avitality has been observed in all skull flaps, which has been significantly increased at the repeated evaluation after 6 months (p < 0.001). Preservation at -23°C (p = 0.006) as well as longer time of storage (p < 0.001) have been identified as prognostic factors for higher rates of bone avitality in a linear mixed regression model.Conclusions Our novel finding shows a clear benefit from a storage at -80° C, which should be carefully considered for the future management and storage of explanted skull flaps. Our analysis further revealed a significant degree of bone avitality, a potential precursor of BFR, also in skull flaps stored for several weeks. To this end, we should reconsider whether the reimplantation of autologous skull flaps instead of synthetic skull flaps is still justified.
Ultrasound-Guided Erector Spinae Plane Block in Elderly Patients Undergoing Total hip arthroplasty (THA): A Prospective Randomized Controlled Study
Authors: Wu Xin-wei , Shen Zhuo-er , Zhou Hong-mei , Li Qi 
Published: June 2023

Abstract
BACKGROUND
Total hip arthroplasty (THA) is associated with considerable postoperative pain among elderly patients. Erector spinae plane(ESP) block is a newly described plane block for post-surgical painmanagement. In this randomized controlled study, we evaluated the efficacy of ultrasound-guided ESP blocks combined with ropivacaine for reducing pain in elderly patients undergoing THA.
METHODS
Patients between 60 and 80 years of age with American Society of Anesthesiologists (ASA) physical status I–III who were scheduled for THA surgery were screened for enrollment. A total of 50 patients were randomized to ESPB group (Group A) receiving ultrasound-guided ESP block and control group receiving no block intervention (Group B). All patients received general anesthesia for surgery. and we recorded their baseline mean arterial pressure (MAP), heart rate(HR), consumption of intraoperative and postoperative opioid, Numerical Rating Scale(NRS) scores, and postoperative adverse reactions during the perioperative period.
RESULTS
The primary outcome was postoperative pain measured by NRS scores. We found that NRS scores were significantly lower in Group A than in Group B in the recovery room and at postoperative 12 and 24 hours (P < 0.05). When the timeline was extended to 48 hours, the NRS scores were no significant difference between this two group(p > 0.05). MAP and HR during surgery but did not find any statistically significant differences between the two groups. The coefficient of variation (CV) of MAP and HR was lower in Group A than in Group B. Total remifentanil consumption in group A was significantly lower than group B. There was no significant difference in complications between these two groups.
CONCLUSIONS
Ultrasound-guided ESP block can improve the stability of MAP and HR in older patients during the surgery, provide effective analgesia in the first 24h after surgery, and reduce opioid requirement, thereby improving the quality of recovery of elderly patients undergoing THA surgery.
Efficient frequency conversion of non-diffracting modes via four-wave mixing in rubidium vapor
Authors: VERMA ONKAR NATH , KANT NITI 
Published: June 2023

Abstract
We investigate the nonlinear frequency conversion of non-diffracting optical modes via non-degenerate four-wave mixing(FWM) process in rubidium vapor. In this process, two strong control fields and a weak probe field mutually interact via afour-level double-Λ type atomic system to produce a new frequency weak Stokes field which is a phase conjugate to probe. We show that any arbitrary mode such as Airy, Bessel, Mathieu, and Weber beams encoded initially in the spatial envelope of probe field are efficiently transferred to FWM Stokes field by satisfying the phase matching condition. Interestingly, we found that the transmitted intensity profiles of Stokes beam are identical to that of probe beam while phase profile is complimentary.The phase conjugation property of output beams is revealed by interfering them with a copropagating plane wave. We further found the fidelity in terms of structural similarity between two transmitted modes of about 99%, which substantiates the high efficiency of FWM process. The efficient transfer and frequency conversion of such diffraction-free beams may have potential applications in nonlinear optical and quantum technologies.
Using the PLS-SEM model to study the factors that affect the willingness of college students to pay for carbon offsets based on a TPB and NAM approach
Authors: Li Changyuan , Liu Xi , Zhang Zhuoya , Li Jiaxi , Zhang Wanxiong , Yang Xin 
Published: June 2023

Abstract

Human influence is very likely the main driver of global climate change according to IPCC AR6 report. Carbon offsets is a new ecological compensation mechanism created against the back-ground of global warming. As college students are an important group in society, the mecha-nism of their willingness to pay for carbon offsets (WTP) is worthy of study. In this study, an in-tegrated theoretical model TPB-NAM is constructed from the perspectives of personal subjective cognition based on the theory of planned behavior (TPB) and the normative activation model (NAM). It also adds two variables, carbon offset cognition (COC) and climate change hazard perception (CCHP). The partial least-squares structural equation model (PLS-SEM) was used to study the factors that affect the WTP of college students. The results show that COC, behavioral attitudes (BAs), subjective norms (SNs), perceived behavioral control (PBC), and personal norms (PNs) have significant positive impacts on college students' WTP. Among them, BAs have the greatest impact on WTP. PNs respectively plays an intermediary role in the four path of AC and SNs affecting BAs, as well as AC and SNs affect WTP. BAs also play a mediating effect in the influence of PNs affect WTP, which is the most obvious.
Edge-based Method for Phenotype Similarity: Enhancing Genetic Disease Diagnosis through Graph-based Representation and Analysis
Authors: Xu Yiteng , Xiong Xu , Xiao Rui 
Published: June 2023

Abstract
Background
The rapid development of high-throughput sequencing technology has revolutionized the diagnosis and understanding of genetic diseases. However, the complexity of clinical phenotypes and high genetic heterogeneity make identifying causative genes a daunting task. In this context, developing accurate and efficient methods to measure the similarity between phenotypes and infer genotype-phenotype associations is crucial for the timely diagnosis and proper management of patients with genetic disorders.
Results
In this study, we proposed an innovative edge-based approach, which we termed PhenoEdge, to measure phenotype similarity, which effectively considers the hierarchical structure of the Human Phenotype Ontology (HPO). Our method demonstrated higher sensitivity than existing methods in both simulated patient datasets and an inhouse dataset of 385 patients with confirmed genetic diseases. Our method showed superior performance in the most clinically relevant simulated dataset containing noise and inaccuracies. Furthermore, the Wilcoxon rank-sum test on the local patient dataset revealed that the sensitivity of our method was significantly better than that of other methods, including the widely used Resnik and Gong methods.
Conclusion
Compared with five existing semantic similarity measures, the edge-based phenotype similarity measurement method demonstrated higher sensitivity and improved accuracy in identifying congenital diseases in clinical settings. This approach can enhance the efficiency of clinical practitioners and holds great promise for future clinical applications. However, further validation with a larger sample size or in real-world clinical scenarios is warranted to fully establish the method's robustness and generalizability.
What helps patients access web-based services in primary care? Free-text analysis of patient responses to the Di-Facto questionnaire
Authors: Khan Nada , Pitchforth Emma , Winder Rachel , Abel Gary , Clark Christopher E. , Cockcroft Emma , Campbell John 
Published: June 2023

Abstract

Background The National Health Service (NHS) and general practice are increasingly adopting digital services. These services can impact both positively and negatively upon patient experiences, but access to digital services is not equal amongst all groups. Within a wider project (the Di-Facto study) our team conducted a patient survey amongst English primary care practices to investigate patient views of what supports uptake and use of web-based services. This paper reports on the analysis of the free-text responses from the patient survey. Methods The Di-Facto patient survey was distributed to practices in eight clinical commissioning groups (CCGs) in England between 2021–2022. We examined free-text responses to two questions relating to access to primary care web-based and support for web-based services. We used qualitative reflexive thematic analysis based on a six-stage process to analyse responses. Results Of the 3051 patients who responded to the Di-Facto survey, 2246 provided a free-text response. We present our findings in two major themes: systems and structures and their impact on use of web-based services, and ‘what works for me’, a description of how respondents described what worked, or did not work in terms of their interactions with web-based services. Respondents described how the technology, such as practice website design, confusion over multiple digital apps, data security and concerns about eConsult offerings impacted on use of web-based services. Respondents described practice level barriers, such as a lack of or inconsistent provision, which prevented optimal use of web-based services. Respondents described personal and technical barriers that impacted on their use of digital services, and described which web-based services worked well for them. Respondents felt that web-based services were not a replacement for face-to-face interactions with a doctor. Conclusions This analysis of free-text responses from a large patient survey highlights the system, practice, and person level barriers and facilitators to use of digital services in primary care. With an increasing push towards digital solutions in NHS primary care, practices should consider the design, rollout and communication of their web-based services to support patient access.
Efficacy of coagulation factor infusion combined with staged surgery in the treatment of moderate hemophilia A combined with kidney stone
Authors: Xu Wenfeng , Liu Deyun , Deng Yaoliang , Dai Yuzong , Guo Fuyou , Wang Xiang 
Published: June 2023

Abstract
Background
To date, there are few reports related to the treatment of patients with hemophilia combined with kidney stones, and there are no standardized treatment protocols for patients with hemophilia combined with kidney stones.
Case presentation:
This case report is of a 27-year-old male patient with hemophilia and left-sided renal calculi. He had severe back pain and bright red hematuria as a result of the disease. On urologic CT we found a left renal pelvic hematoma. The patient was treated with coagulation factors and staged surgery, and his kidney stones have now been cured.
Conclusions
Patients with hemophilia A are at high-risk for operation and the expense of transfusion of clotting factors is expensive, so strict indications for operation must be followed, otherwise operative treatment should be avoided. When dealing with patients with moderate hemophilia A combined with kidney stones, if they have ureteral obstruction, back pain, bright red hematuria, etc. If the patient's condition is good and medical conditions allow, operative treatment should be performed as soon as possible. If a patient has a pelvic hematoma and severe hematuria, in addition to the need to give the patient a clotting factor infusion, we can stage the surgery to reduce the risk of the associated surgery. The first period is to drain the pelvic hematoma to relieve the patient's hard and improve the patient's renal function, and also to prepare for the second period of lithotripsy. After the patient's general condition improves, then we can perform lithotripsy and try to remove the lithotripsy to reduce the bleeding caused by lithotripsy and ureteral stent.
The psychological experience of advanced maternal age raising a second child: a qualitative study
Authors: Ge Yurong , Zhang Xin , Li Juanjuan , Zheng Jingyuan , Zeng Fen , Kitili Ike , Chen Qianyi 
Published: June 2023

Abstract
Objective:
To explore the psychological changes that may take place in older women with a second child from conception to birth. The study has several aims, including the provision of protection of the overall health level of these women; the maintenance of women's position in the labor market; and the promotion of theimplementation of the "comprehensive two-child" policy.
Methods:
Semi-open interviews were conducted on older women pregnant with their second child. These women were recruited from three tertiary hospitals in one province in western China. The interview content was analyzed using the qualitative research method of Japanese scholar Kinoshita Yasuhito's revised version of grounded theory.
Results:
Four themes were extracted from the psychological experience of the older women with a second child; these were: Life choice; The mixed feelings of motherhood; The backbone of a family; Second children should be cautious.
Conclusion:
older women with a second child face physical, psychological, life, and other pressures. To ensure the implementation of the "comprehensive two-child" policy, the Chinese government should speed up the establishment of a fertility support system, including education, medical investment, women’s employment rights protection, and pension burden. Establish and improve relevant policies.