bluuebunny/crossref_metadata_2025_split huggingface dataset:
Challenges and strategies for conducting research in primary health care practice: an integrative review
Authors: Bonfim Daiana , Belotti Lorrayne , de Almeida Leticia Yamawaka , Eshriqui Ilana , Velasco Sofia Rafaela Maito , Monteiro Camila Nascimento , Jantsch Adelson Guaraci 
Published: December 2023

Abstract
Background
Providing accessible and high-quality patient-centered healthcare remains a challenge in many countries, despite global efforts to strengthen primary health care (PHC). Research and knowledge management are integral to enhancing PHC, facilitating the implementation of successful strategies, and promoting the use of evidence-based practices. Practice-based research in primary care (PC-PBR) has emerged as a valuable approach, with its external validity to diverse PHC settings, making it an effective means of translating research findings into professional practice.

Objective
To identify challenges and strategies for conducting practice-based research in primary health care services.

Method
An integrative literature review was conducted by searching the PubMed, Embase, Scopus, Web of Science, and Lilacs databases. The research question, guided by the PICo framework, directed the execution of study selection and data extraction. Data analysis followed the RAdAR method's three phases: pre-analysis, data analysis, and interpretation of results.

Results
Out of 440 initially identified articles, 26 met the inclusion criteria. Most studies were conducted in high-income countries, primarily the United States. The challenges and strategies for PC-PBR were categorized into six themes: research planning, infrastructure, engagement of healthcare professionals, knowledge translation, the relationship between universities and health services, and international collaboration. Notable challenges included research planning complexities, lack of infrastructure, difficulties in engaging healthcare professionals, and barriers to knowledge translation. Strategies underscore the importance of adapting research agendas to local contexts, providing research training, fostering stakeholder engagement, and establishing practice-based research networks.

Conclusion
The challenges encountered in PC-PBR are consistent across various contexts, highlighting the need for systematic, long-term actions involving health managers, decision-makers, academics, diverse healthcare professionals, and patients. This approach is essential to transform primary care, especially in low- and middle-income countries, into an innovative, comprehensive, patient-centered, and accessible healthcare system. By addressing these challenges and implementing the strategies, PC-PBR can play a pivotal role in bridging the gap between research and practice, ultimately improving patient care and population health.
DNA polymerases in precise and predictable CRISPR/Cas9-mediated chromosomal rearrangements
Authors: Mehryar Mohammadreza M. , Shi Xin , Li Jingwei , Wu Qiang 
Published: December 2023

Abstract
Background
Recent studies have shown that, owning to its cohesive cleavage, Cas9-mediated CRISPR gene editing outcomes at junctions of chromosomal rearrangements or DNA-fragment editing are precise and predictable; however, the underlying mechanisms are poorly understood due to lack of suitable assay system and analysis tool.

Results
Here we developed a customized computer program to take account of staggered or cohesive Cas9 cleavage and to rapidly process large volumes of junctional sequencing reads from chromosomal rearrangements or DNA-fragment editing, including DNA-fragment inversions, duplications, and deletions. We also established a sensitive assay system using HPRT1 and DCK as reporters for cell growth during DNA-fragment editing by Cas9 with dual sgRNAs and found prominent large resections or long deletions at junctions of chromosomal rearrangements. In addition, we found that knockdown of PolQ (encoding Polθ polymerase), which has a prominent role in theta-mediated end joining (TMEJ) or microhomology-mediated end joining (MMEJ), results in increased large resections but decreased small deletions. We also found that the mechanisms for generating small deletions of 1bp and >1bp during DNA-fragment editing are different with regard to their opposite dependencies on Polθ and Polλ (encoded by the PolL gene). Specifically, Polθ suppresses 1bp deletions but promotes >1bp deletions, whereas Polλ promotes 1bp deletions but suppresses >1bp deletions. Finally, we found that Polλ is the main DNA polymerase responsible for fill-in of the 5′ overhangs of staggered Cas9 cleavage ends.

Conclusions
These findings contribute to our understanding of the molecular mechanisms of CRISPR/Cas9-mediated DNA-fragment editing and have important implications for controllable, precise, and predictable gene editing.
One-year psychophysical evaluation of COVID-19-induced olfactory disorders: a prospective cohort study
Authors: Schepens Emma J. A. , Boek Wilbert M. , Boesveldt Sanne , Stokroos Robert J. , Stegeman Inge , Kamalski Digna M. A. 
Published: December 2023

Abstract
Background
Olfactory disorders are common in COVID-19. While many patients recover within weeks, a notable number of patients suffer from prolonged olfactory disorders. Much research has focused on the acute phase of olfactory disorders in COVID-19; however, there is still inconsistency regarding the prognosis. We aim to assess both objective and subjective olfactory function in patients with persisting olfactory disorders following COVID-19, 1 year after diagnosis.

Methods
We objectively measured olfactory function in 77 patients who initially had COVID-19-induced smell disorders, 1 year after confirmed diagnosis. These patients previously underwent two objective measurements at approximately 3 and 6 months after COVID-19, in the context of the COCOS trial (COrticosteroids for COvid-19-induced loss of Smell). The main outcome measurement was TDI score (threshold-discrimination-identification) on Sniffin’ Sticks Test (SST). Secondary outcomes included objective gustatory function on Taste Strip Test (TST), self-reported olfactory, gustatory and trigeminal function on a visual analogue scale (VAS) and outcomes on questionnaires about quality of life, and nasal symptoms.

Results
The findings of this study show that 1 year following COVID-19, the median TDI score increased to 30.75 (IQR 27.38–33.5), regarded as normosmia. The median TDI score started at 21.25 (IQR 18.25–24.75) at baseline and increased to 27.5 (IQR 23.63–30.0) at 6 months following COVID-19. The increase of 9.5 points on the TDI score between baseline and 1 year after COVID-19 marks a clinically relevant improvement. Regarding the self-reported VAS score (1–10) on sense of smell, it increased from 1.2 (IQR 0.4–3.0) at baseline to 3.2 (IQR 1.4–6.0) at 6 months and further improved up to 6.1 (IQR 2.7–7.5) after 1 year. Objective gustatory function increased with 2 points on TST a year after diagnosis. Self-reported olfactory, gustatory, and trigeminal functions also improved over time, as did quality of life.

Conclusions
Objective and self-reported olfactory function continued to improve 1 year after COVID-19. The median TDI score of 30.75 (IQR 27.38–33.5) is regarded as normosmia, which is a favorable outcome. However, the rate of improvement on TDI score reduces over time.
Selective extraction of recombinant membrane proteins from Hansenula polymorpha by pulsed electric field and lytic enzyme pretreatment
Authors: Ganeva Valentina , Kranz Andreas 
Published: December 2023

Abstract
Background
In yeast, recombinant membrane proteins including viral scaffold proteins used for the formation of enveloped Virus-like particles (eVLPs) typically accumulate intracellularly. Their recovery is carried out by mechanical disruption of the cells, often in combination with detergent treatment. Cell permeabilization is an attractive alternative to mechanical lysis because it allows for milder and more selective recovery of different intracellular products.

Results
Here, we present a novel approach for extraction of integral membrane proteins from yeast based on cell envelope permeabilization through a combination of pulsed electric field and lytic enzyme pretreatment of the cells. Our primary experiments focused on Hansenula polymorpha strain #25-5 co-expressing the integral membrane small surface protein (dS) of the duck hepatitis B virus and a fusion protein of dS with a trimer of a Human papillomavirus (HPV) L2-peptide (3xL2-dS). Irreversible plasma membrane permeabilization was induced by treating the cell suspension with monopolar rectangular pulses using a continuous flow system. The permeabilized cells were incubated with lyticase and dithiothreitol. This treatment increased the cell wall permeability, resulting in the release of over 50% of the soluble host proteins without causing significant cell lysis. The subsequent incubation with Triton X-100 resulted in the solubilization and release of a significant portion of 3xL2-dS and dS from the cells. By applying two steps: (i) brief heating of the cells before detergent treatment, and (ii) incubation of the extracts with KSCN, an 80% purity on the protein level has been achieved. Experiments performed with H. polymorpha strain T#3-3, co-expressing dS and the fusion protein EDIIIWNV-dS consisting of dS and the antigen from the West Nile virus (WSV), confirmed the applicability of this approach for recovering dS. The treatment, optimal for solubilization of 3xL2-dS and a significant part of dS, was not effective in isolating the fused protein EDIIIWNV-dS from the membranes, resulting in its retention within the cells.

Conclusions
This study presents an alternative approach for the recovery and partial purification of viral membrane proteins expressed in H. polymorpha. The factors influencing the effectiveness of this procedure and its potential use for the recovery of other integral membrane proteins are discussed.
Tumour cells can escape antiproliferative pressure by interferon-β through immunoediting of interferon receptor expression
Authors: Hiebinger Felix , Kudulyte Aiste , Chi Huanting , Burbano De Lara Sebastian , Ilic Doroteja , Helm Barbara , Welsch Hendrik , Dao Thi Viet Loan , Klingmüller Ursula , Binder Marco 
Published: December 2023

AbstractType I interferons (IFNs) play a central role not only in innate immunity against viral infection, but also in the antitumour response, e.g. through a direct impact on cell proliferation. Particularly for cancer arising in the context of chronic inflammation, constant exposure to IFNs may constitute a strong selective pressure during tumour evolution. Expansion of neoplastic subclones resistant to the antiproliferative effects of IFNs may contribute to immunoediting of tumours, leading to more aggressive disease. Experimental evidence for this development of IFN-insensitivity has been scarce and its molecular mechanism is unclear. In this study we demonstrate that six weeks exposure of cells to IFN-β in vitro reduces their sensitivity to its antiproliferative effects, and that this phenotype was stable for up to four weeks. Furthermore, we observed substantial differences in cellular sensitivity to growth inhibition by IFN-β in a panel of ten different liver cancer cell lines, most prominently in a pair of highly dedifferentiated cell lines, and least in cells from well-differentiated tumours. In both, long-term IFN selection and in dedifferentiated tumour cell lines, we found IFNAR2 expression to be substantially reduced, suggesting the receptor complex to be a sensitive target amenable to immunoediting. Beyond new insights into possible molecular processes in tumour evolution, these findings might prove valuable for the development of biomarkers allowing to stratify tumours for their sensitivity to IFN treatment in the context of patient tailored therapies.
A critical review and analysis of the context, current burden, and application of policy to improve cancer equity in Ghana
Authors: Tuck Chloe Zabrina , Cooper Richard , Aryeetey Richmond , Gray Laura A , Akparibo Robert 
Published: December 2023

Abstract
Background
Cancer causes a major disease burden worldwide. This is increasingly being realised in low and middle-income countries, which account disproportionately for preventable cancer deaths. Despite the World Health Organization calling for governments to develop policies to address this and alleviate cancer inequality, numerous challenges in executing effective cancer policies remain, which require consideration of the country-specific context. As this has not yet been considered in Ghana, the aim of this review was to bring together and critique the social-environmental, health policy and system factors to identifying opportunities for future health policies to reduce cancer burden in the Ghanian context. A critical policy-focused review was conducted to bring together and critique the current health systems context relating to cancer in Ghana, considering the unmet policy need, health system and social factors contributing to the burden and policy advances related to cancer.

Conclusion
The findings highlight the changing burden of cancer in Ghana and the contextual factors within the socio-ecosystem that contribute to this. Policies around expanding access to and coverage of services, as well as the harmonization with medical pluralism have potential to improve outcomes and increase equity but their implementation and robust data to monitor their impact pose significant barriers.
Ozone alleviates MSU-induced acute gout pain via upregulating AMPK/GAS6/MerTK/SOCS3 signaling pathway
Authors: Fan Wen , Liu Chong , Chen Dacai , Xu Chenjie , Qi Xiuting , Zhang Ailin , Zhu Xuexian , Liu Yujie , Wang Lei , Hao Lanxiang , Liu Wen-Tao , Hu Liang 
Published: December 2023

Abstract
Background
Gout pain seriously affects the quality of patients' life. There is still no effective treatment. The inflammatory response is the main mechanism of gout. Here, we found that ozone can reduce the inflammatory reaction in the joints of gouty mice and relieve gout pain, and we further explore its protective mechanism.

Methods
MSU was used to establish the gouty mice model. Nociception was assessed by Von Frey hairs. Cell signaling assays were performed by western blotting and immunohistochemistry. The mouse leukemia cells of monocyte macrophage line RAW264.7 were cultured to investigate the effects of ozone administration on macrophage.

Results
Ozone reduced inflammation, relieved gout pain and improved the paw mean intensity and duty cycle of the gouty mice. Ozone increased the phosphorylation of AMP-activated protein kinase (AMPK), induced suppressor of cytokine signaling 3 (SOCS3) expression and inhibited metallopeptidase 9 (MMP9) expression. In vivo, ozone activated AMPK to induce Gas6 release, and upregulated MerTK/SOCS3 signaling pathway to reduce inflammation in mouse macrophage line RAW264.7. Inhibitors of AMPK and MerTK, respectively abolished the analgesic and anti-inflammatory effects of ozone in vivo and in vitro. Gas6 knockout cancelled the protectively effects of ozone on gout pain and the paw mean intensity and duty cycle of gouty mice. Additionally, the level of Gas6 and protein S in plasma of patients with hyperuricemia was significantly higher than that of healthy contrast group.

Conclusion
Ozone reduces inflammation and alleviates gout pain by activating AMPK to up-regulate Gas6/MerTK/SOCS3 signaling pathway.
Efficacy of aquatic exercise in chronic musculoskeletal disorders: a systematic review and meta-analysis of randomized controlled trials
Authors: Wang Tianyue , Wang Jiamin , Chen Yuheng , Ruan Yanmin , Dai Senjie 
Published: December 2023

Abstract
Background
Aquatic exercise (AE) is becoming ever more popular as a physical therapy, while it is unclear what precise improvements it will produce and how effective it will be in comparison with other non-surgical therapies. The study aimed to assess whether AE positively impacts chronic musculoskeletal disorder patients in terms of pain, physical function, and quality of life.

Methods
PRISMA guidelines were followed, and our study protocol was published online at PROSPERO under registration number CRD42023417411. We searched PubMed, Embase, Web of Science, and Cochrane library databases for English-language articles published before April 11, 2023, including studies from all relevant randomized controlled trials (RCTs). After screening, we ultimately included 32 RCTs with a total of 2,200 participants. We also performed subgroup analyses for all included studies. This meta-analysis calculated standardized mean difference (SMD) with 95% confidence interval (CI), and the variance was estimated using a random-effects model. The quality of the included studies was assessed by using the Cochrane collaborative "risk of bias" assessment tool (version 2.0). Thus ensuring that the literature included is of high quality.

Results
This meta-analysis included 32 trials with 2,200 participants; these patients were all between the ages of 38–80. The study showed that compared to the no exercise (NE) group, patients in the AE group experienced a remarkable reduction in pain (SMD: -0.64, P < 0.001), a significant increase in physical function (SMD: 0.62, P < 0.001), and a statistically significant improvement in quality of life (SMD: −0.64, P < 0.001). When compared to land-based exercise (LE), AE significantly relieves patients' pain (SMD: −0.35, P = 0.03).

Conclusions
This is the first systematic review and meta-analysis to study whether AE could improve chronic musculoskeletal disorders. The evidence suggests that AE benefits pain, physical function, and quality of life in adults with chronic musculoskeletal conditions compared to NE. Furthermore, when compared to LE, AE continues to provide a better improvement in patient pain. More long-term clinical trials are needed to confirm AE's positive effects and improvement mechanisms and the more existential advantages compared to LE.
Demographic, clinical, and paraclinical features of patients operated with the diagnosis of acute descending necrotizing mediastinitis: a retrospective study in Southern Iran
Authors: Ranjbar Keivan , Shahriarirad Reza , Ebrahimi Kamyar , Amirian Armin , Karoobi Mohamadreza , Mardani Parviz , Erfani Amirhossein , Fallahi Mohammad Javad , Ketabchi Farzaneh , Ziaian Bizhan 
Published: December 2023

Abstract
Introduction
Descending necrotizing mediastinitis (DNM) is a type of acute mediastinitis that is rarely reported but is regarded as a fatal disease despite improvements in technological methods and antibiotic therapies. We aimed to determine the demographic, clinical, and paraclinical features of patients diagnosed with acute DNM.

Methods
In this retrospective study, patients’ hospital records with a diagnosis of DNM admitted to the Namazi hospital in southern Iran during 18 years (2002–2019) were reviewed. Demographic and clinical features were recorded and subsequently analyzed via SPSS 22.

Results
Out of 67 mediastinitis patients, 25 (37.3%) were diagnosed as DNM with an average age of 37.2 ± 16.7 years, and 68% were male. Regarding etiology, 52.0% were due to neck infection. Based on the technique of surgery, 52% of the patients underwent the combined method, which was mostly among type I and IIA DNM, while thoracotomy was mostly performed on type IIB DNM (P = 0.08). Based on the incision, type IIA and IIB had the highest frequency of thoracotomy and cervicothoracic incisions (P = 0.02 and 0.002). Puss discharge was significantly lower in type I DNM (P = 0.01). Based on the presenting symptoms of our patients, the majority (72.0%) had a chief complaint of neck pain, followed by chills and fever (48%). There were no reports of mortality during our short-term follow-up.

Conclusion
We report one of the largest retrospective studies of DNM patients in our referral center, with a high prevalence of the disease among younger populations, especially under 40 years. The method of treatment should be chosen based on the extent of infection and can be limited to neck exploration in upper mediastinal infections, though thoracic or combined approach in more broad infections.
Characterization of MicroRNA expression profiles in the ovarian tissue of goats during the sexual maturity period
Authors: Wang Yanyan , Wang Jianmin , Li Qing , Xuan Rong , Guo Yanfei , He Peipei , Chao Tianle 
Published: December 2023

Abstract
Background
The ovary is an important reproductive organ in mammals, and its development directly affects the sexual maturity and reproductive capacity of individuals. MicroRNAs (miRNAs) are recognized as regulators of reproductive physiological processes in various animals and have been shown to regulate ovarian development through typical targeting and translational repression. However, little is known about the regulatory role of miRNAs in ovarian tissue development during sexual maturity in goats. To comprehensively profile the different physiological stages of sexual maturation in goats, we performed small-RNA sequencing of ovarian tissue samples collected at four specific time points (1 day after birth (D1), 2 months old (M2), 4 months old (M4), and 6 months old (M6)). In addition, we used ELISAs to measure serum levels of reproductive hormones to study their temporal changes.

Results
The results showed that serum levels of gonadotropin-releasing hormone, follicle-stimulating hormone, luteinizing hormone, oestradiol, progesterone, oxytocin, and prolactin were lower in goats at the D1 stage than at the other three developmental stages (P < 0.05). The secretion patterns of these seven hormones show a similar trend, with hormone levels reaching their peaks at 4 months of age. A total of 667 miRNAs were detected in 20 libraries, and 254 differentially expressed miRNAs and 3 groups of miRNA clusters that had unique expression patterns were identified (|log2-fold change|> 1, FDR < 0.05) in the 6 comparison groups. RT‒qPCR was employed to confirm that the expression pattern of the 15 selected miRNAs was consistent with the Illumina sequencing results. Gene ontology analyses revealed significant enrichment of GO terms such as cell proliferation regulation, epithelial cell development, and amino acid transport, as well as important signaling pathways including the MAPK signaling pathway, the PI3K-Akt signaling pathway, and the oestrogen signaling pathway. Further miRNA‒mRNA regulation network analysis revealed that 8 differentially expressed miRNAs (chi-miR-1343, chi-miR-328-3p, chi-miR-877-3p, chi-miR-296-3p, chi-miR-128-5p, chi-miR-331-3p, chi-miR-342-5p and chi-miR-34a) have important regulatory roles in ovarian cell proliferation, hormone secretion and metabolism-related biological processes.

Conclusions
Overall, our study investigated the changes in serum hormone and miRNA levels in the ovaries. These data provide a valuable resource for understanding the molecular regulatory mechanisms of miRNAs in ovarian tissue during the sexual maturity period in goats.