bluuebunny/crossref_metadata_2025 huggingface dataset:
B-to-A transition in target DNA during retroviral integration
Authors: Jóźwik Ilona K , Li Wen , Zhang Da-Wei , Wong Doris , Grawenhoff Julia , Ballandras-Colas Allison , Aiyer Sriram , Cherepanov Peter , Engelman Alan N , Lyumkis Dmitry
Published: August 2022

Integration into host target DNA (tDNA), a hallmark of retroviral replication, is mediated by the intasome, a multimer of integrase (IN) assembled on viral DNA (vDNA) ends. To ascertain aspects of tDNA recognition during integration, we have solved the 3.5 Å resolution cryo-EM structure of the mouse mammary tumor virus (MMTV) strand transfer complex (STC) intasome. The tDNA adopts an A-like conformation in the region encompassing the sites of vDNA joining, which exposes the sugar-phosphate backbone for IN-mediated strand transfer. Examination of existing retroviral STC structures revealed conservation of A-form tDNA in the analogous regions of these complexes. Furthermore, analyses of sequence preferences in genomic integration sites selectively targeted by six different retroviruses highlighted consistent propensity for A-philic sequences at the sites of vDNA joining. Our structure additionally revealed several novel MMTV IN-DNA interactions, as well as contacts seen in prior STC structures, including conserved Pro125 and Tyr149 residues interacting with tDNA. In infected cells, Pro125 substitutions impacted the global pattern of MMTV integration without significantly altering local base sequence preferences at vDNA insertion sites. Collectively, these data advance our understanding of retroviral intasome structure and function, as well as factors that influence patterns of vDNA integration in genomic DNA.
TGS1 impacts snRNA 3′-end processing, ameliorates survival motor neuron-dependent neurological phenotypes in vivo and prevents neurodegeneration
Authors: Chen Lu , Roake Caitlin M , Maccallini Paolo , Bavasso Francesca , Dehghannasiri Roozbeh , Santonicola Pamela , Mendoza-Ferreira Natalia , Scatolini Livia , Rizzuti Ludovico , Esposito Alessandro , Gallotta Ivan , Francia Sofia , Cacchione Stefano , Galati Alessandra , Palumbo Valeria , Kobin Marie A , Tartaglia Gian Gaetano , Colantoni Alessio , Proietti Gabriele , Wu Yunming , Hammerschmidt Matthias , De Pittà Cristiano , Sales Gabriele , Salzman Julia , Pellizzoni Livio , Wirth Brunhilde , Di Schiavi Elia , Gatti Maurizio , Artandi Steven E , Raffa Grazia D
Published: August 2022

Trimethylguanosine synthase 1 (TGS1) is a highly conserved enzyme that converts the 5′-monomethylguanosine cap of small nuclear RNAs (snRNAs) to a trimethylguanosine cap. Here, we show that loss of TGS1 in Caenorhabditis elegans, Drosophila melanogaster and Danio rerio results in neurological phenotypes similar to those caused by survival motor neuron (SMN) deficiency. Importantly, expression of human TGS1 ameliorates the SMN-dependent neurological phenotypes in both flies and worms, revealing that TGS1 can partly counteract the effects of SMN deficiency. TGS1 loss in HeLa cells leads to the accumulation of immature U2 and U4atac snRNAs with long 3′ tails that are often uridylated. snRNAs with defective 3′ terminations also accumulate in Drosophila Tgs1 mutants. Consistent with defective snRNA maturation, TGS1 and SMN mutant cells also exhibit partially overlapping transcriptome alterations that include aberrantly spliced and readthrough transcripts. Together, these results identify a neuroprotective function for TGS1 and reinforce the view that defective snRNA maturation affects neuronal viability and function.
Strand switching mechanism of Pif1 helicase induced by its collision with a G-quadruplex embedded in dsDNA
Authors: Valle-Orero Jessica , Rieu Martin , Tran Phong Lan Thao , Joubert Alexandra , Raj Saurabh , Allemand Jean-François , Croquette Vincent , Boulé Jean-Baptiste
Published: August 2022

G-rich sequences found at multiple sites throughout all genomes may form secondary structures called G-quadruplexes (G4), which act as roadblocks for molecular motors. Among the enzymes thought to process these structures, the Pif1 DNA helicase is considered as an archetypical G4-resolvase and its absence has been linked to G4-related genomic instabilities in yeast. Here we developed a single-molecule assay to observe Pif1 opening a DNA duplex and resolving the G4 in real time. In support of former enzymological studies, we show that the helicase reduces the lifetime of G4 from hours to seconds. However, we observe that in the presence of a G4, Pif1 exhibits a strong strand switching behavior, which can lead to Pif1 escaping G4 resolution, depending on the structural context surrounding the substrate. This behavior is also detected in the presence of other roadblocks (LNA or RNA). We propose that the efficiency of Pif1 to remove a roadblock (G4 or other) is affected by its strand switching behavior and depends on the context surrounding the obstacle. We discuss how this switching behavior may explain several aspects of Pif1 substrate preference and affect its activity as a G4 resolvase in vivo.
Molecular dynamics of DNA translocation by FtsK
Authors: Pajak Joshua , Arya Gaurav
Published: August 2022

The bacterial FtsK motor harvests energy from ATP to translocate double-stranded DNA during cell division. Here, we probe the molecular mechanisms underlying coordinated DNA translocation in FtsK by performing long timescale simulations of its hexameric assembly and individual subunits. From these simulations we predict signaling pathways that connect the ATPase active site to DNA-gripping residues, which allows the motor to coordinate its translocation activity with its ATPase activity. Additionally, we utilize well-tempered metadynamics simulations to compute free-energy landscapes that elucidate the extended-to-compact transition involved in force generation. We show that nucleotide binding promotes a compact conformation of a motor subunit, whereas the apo subunit is flexible. Together, our results support a mechanism whereby each ATP-bound subunit of the motor conforms to the helical pitch of DNA, and ATP hydrolysis/product release causes a subunit to lose grip of DNA. By ordinally engaging and disengaging with DNA, the FtsK motor unidirectionally translocates DNA.
Biochemical and structural characterization of hepatitis A virus 2C reveals an unusual ribonuclease activity on single-stranded RNA
Authors: Chen Pu , Wojdyla Justyna Aleksandra , Colasanti Ombretta , Li Zhijian , Qin Bo , Wang Meitian , Lohmann Volker , Cui Sheng
Published: August 2022

The HAV nonstructural protein 2C is essential for virus replication; however, its precise function remains elusive. Although HAV 2C shares 24–27% sequence identity with other 2Cs, key motifs are conserved. Here, we demonstrate that HAV 2C is an ATPase but lacking helicase activity. We identified an ATPase-independent nuclease activity of HAV 2C with a preference for polyuridylic single-stranded RNAs. We determined the crystal structure of an HAV 2C fragment to 2.2 Å resolution, containing an ATPase domain, a region equivalent to enterovirus 2C zinc-finger (ZFER) and a C-terminal amphipathic helix (PBD). The PBD of HAV 2C occupies a hydrophobic pocket (Pocket) in the adjacent 2C, and we show the PBD–Pocket interaction is vital for 2C functions. We identified acidic residues that are essential for the ribonuclease activity and demonstrated mutations at these sites abrogate virus replication. We built a hexameric-ring model of HAV 2C, revealing the ribonuclease-essential residues clustering around the central pore of the ring, whereas the ATPase active sites line up at the gaps between adjacent 2Cs. Finally, we show the ribonuclease activity is shared by other picornavirus 2Cs. Our findings identified a previously unfound activity of picornavirus 2C, providing novel insights into the mechanisms of virus replication.
Tacrolimus induces a pro-fibrotic response in donor-derived human proximal tubule cells dependent on common variants of theCYP3A5andABCB1genes
Authors: Knops Noël , Ramazani Yasaman , De Loor Henriëtte , Goldschmeding Roel , Nguyen Tri Q , van den Heuvel Lambert P , Levtchenko Elena , Kuypers Dirk J
Published: August 2022

Background

Common genetic variants of the enzymes and efflux pump involved in tacrolimus disposition have been associated with calcineurin inhibitor nephrotoxicity, but their importance is unclear because of the multifactorial background of renal fibrosis. This study explores the pro-fibrotic response of tacrolimus exposure in relation to the differential capacity for tacrolimus metabolism in proximal tubule cells (PTCs) with a variable (pharmaco)genetic background.

Methods

PTCs were obtained from protocol allograft biopsies with different combinations of CYP3A5 and ABCB1 variants and were incubated with tacrolimus within the concentration range found in vivo. Gene and protein expression, CYP3A5 and P-glycoprotein function, and tacrolimus metabolites were measured in PTC. Connective tissue growth factor (CTGF) expression was assessed in protocol biopsies of kidney allograft recipients.

Results

PTCs produce CTGF in response to escalating tacrolimus exposure, which is approximately 2-fold higher in cells with the CYP3A5*1 and ABCB1 TT combination in vitro. Increasing tacrolimus exposure results in relative higher generation of the main tacrolimus metabolite {13-O-desmethyl tacrolimus [M1]} in cells with this same genetic background. Protocol biopsies show a larger increase in in vivo CTGF tissue expression over time in TT vs. CC/CT but was not affected by the CYP3A5 genotype.

Conclusions

Tacrolimus exposure induces a pro-fibrotic response in a PTC model in function of the donor pharmacogenetic background associated with tacrolimus metabolism. This finding provides a mechanistic insight into the nephrotoxicity associated with tacrolimus treatment and offers opportunities for a tailored immunosuppressive treatment.
Tenofovir-Diphosphate in Dried Blood Spots vs Tenofovir in Urine/Plasma for Oral Preexposure Prophylaxis Adherence Monitoring
Authors: Niu Xin , Kubiak Rachel W , Siriprakaisil Oraphan , Klinbuyaem Virat , Sukrakanchana Pra ornsuda , Cressey Ratchada , Okochi Hideaki , Gandhi Monica , Cressey Tim R , Drain Paul K
Published: August 2022

Background

Tenofovir-diphosphate (TFV-DP) measured in dried blood spots (DBS) and tenofovir (TFV) measured in urine/plasma have been used to measure TFV-based oral pre-exposure prophylaxis (PrEP) adherence. However, there are limited data comparing these 3 metrics and their appropriate use for PrEP adherence monitoring.

Methods

We collected DBS, urine, and plasma samples from HIV-negative adults randomized to a low (2 doses/week), moderate (4 doses/week), or perfect (7 doses/week) adherence group (via directly observed therapy) of tenofovir disoproxil fumarate/emtricitabine (TDF/FTC) for 6 weeks, followed by a 4-week washout phase. Drug concentrations were measured using liquid chromatography tandem mass spectrometry. Linear mixed-effects modeling was used to examine associations between drug concentrations and dosing time.

Results

Among 28 participants, the median age was 33 years, and 12 (43%) were female. At steady state, 25th percentile TFV-DP concentrations were 466, 779, and 1375 fmol/3 mm punch in the low, moderate, and perfect adherence group, respectively. Correlation was stronger between quantifiable TFV-DP and plasma TFV (r = 0.65; P < .01) than between TFV-DP and urine TFV (r = 0.50; P < .01). Among all participants, each additional week of cumulative dosing on average led to a mean increase of 158 fmol/3 mm punch (P < .001) in TFV-DP during the dosing phase. Each additional day after the last dose was associated with 43 fmol/3 mm punch lower TFV-DP (P = .07).

Conclusions

TFV-DP levels in DBS provide valuable insight into both dosing recency and cumulative doses from variable adherence patterns. Our observed benchmark TFV-DP concentrations were slightly higher than prior predicted estimates based on convenience samples.
Comparing the Efficacy and Safety of Gemcitabine plus Nab-Paclitaxel versus Gemcitabine Alone in Older Adults with Unresectable Pancreatic Cancer
Authors: Kobayashi Satoshi , Suzuki Motoko , Ueno Makoto , Maruki Yuta , Okano Naohiro , Todaka Akiko , Ozaka Masato , Tsuji Kunihiro , Shioji Kazuhiko , Doi Keitaro , Kojima Yasushi , Tsumura Hidetaka , Tanaka Kazunari , Higuchi Hajime , Kawabe Ken , Imaoka Hiroshi , Yamashita Tatsuya , Miwa Haruo , Nagano Hiroaki , Arima Shiho , Hayashi Hideyuki , Naganuma Atsushi , Yamaguchi Hironori , Hisano Terumasa , Umemoto Kumiko , Ishii Shuji , Nakashima Koji , Suzuki Rei , Kitano Yohei , Misumi Toshihiro , Furuse Junji , Ishii Hiroshi
Published: August 2022

Background

Gemcitabine plus nab-paclitaxel (GnP) has been a standard treatment for unresectable pancreatic cancer (uPC); however, the current treatment status and usefulness in older adults with uPC remain unclear. Therefore, we aimed to investigate the patient background and compare the efficacy and safety of GnP versus other treatments in older adults with uPC.

Patients and Methods

In this prospective observational study, we enrolled 233 eligible patients aged ≥76 years with pathologically proven, clinically uPC, and no history of chemotherapy from 55 Japanese centers during September 2018-September 2019. The main endpoints were overall survival (OS), progression-free survival (PFS), and safety. Geriatric assessments were performed upon registration and after 3 months. To adjust for confounders, we conducted propensity score-matched analyses.

Results

GnP, gemcitabine alone (Gem), best supportive care, and other therapies were administered to 116, 72, 16, and 29 patients, respectively. In the propensity score-matched analysis, 42 patients each were selected from the GnP and Gem groups. The median OS was longer in the GnP group than in the Gem group (12.2 vs. 9.4 months; hazard ratio [HR], 0.65; 95% CI, 0.37-1.13). The median PFS was significantly longer in the GnP group than in the Gem group (9.2 vs. 3.7 months; HR, 0.38; 95% CI, 0.23-0.64). The incidence of severe adverse events was higher with GnP than with Gem; however, the difference was not significant.

Conclusion

GnP is more efficacious than Gem in patients aged ≥76 years with uPC despite demonstrating a higher incidence of severe adverse events.
Serum biomarkers in prednisolone-treated hand osteoarthritis patients
Authors: van de Stadt Lotte A , Kroon Féline P B , Thudium Christian F , Bay-Jensen Anne C , Kloppenburg Margreet
Published: August 2022

Objectives

To investigate whether biomarkers are modulated by prednisolone treatment in patients with hand OA and whether they can predict response to prednisolone.

Methods

Biomarkers reflecting tissue turnover and inflammation [aggrecanase-derived neoepitope of arggecan (ARGS), MMP-derived neoepitope of type I collagen (C1M), MMP-derived neoepitope of type III collagen (C3M), marker of true type V collagen formation (PROC5), MMP-derived neoepitope of CRP (CRPM), citrullinated vimentin fragment (VICM), high-sensitivity (hsCRP)] were measured in sera from 78 patients with painful inflammatory hand OA, who were randomized between prednisolone or placebo treatment. Association of baseline biomarker levels with disease characteristics [visual analogue scale (VAS) pain, synovial thickening ultrasonography sum score and erosive OA] and OMERACT-Osteoarthritis Research Society International (OARSI) response after 6 weeks were analysed with linear or logistic regression and adjusted for age, BMI and sex. Change in biomarker levels after 6 weeks was assessed with linear regression adjusted for baseline biomarker levels, age, BMI and sex.

Results

For all patients (mean age 64 years, 79% female), there were no associations between biomarker levels and VAS finger pain or synovial thickening score at baseline. Patients with erosive hand OA had higher levels of C1M and hsCRP [adjusted geometric mean ratio 1.24 (95% CI 1.03, 1.49) and 1.91 (1.19, 3.06), respectively]. Biomarker levels did not decrease over time. There was no association between baseline biomarkers levels and OARSI response, except for CRPM [geometric mean ratio of 0.88 (0.77, 1.00)].

Conclusion

Erosive disease was associated with higher levels of C1M and hsCRP. Biomarker levels were not influenced by treatment with prednisolone. Current biomarkers were not associated with response to prednisolone in hand OA.
Human dermal fibroblast-derived exosomes induce macrophage activation in systemic sclerosis
Authors: Bhandari Rajan , Yang Heetaek , Kosarek Noelle N , Smith Avi E , Garlick Jonathan A , Hinchcliff Monique , Whitfield Michael L , Pioli Patricia A
Published: August 2022

Objectives

Prior work demonstrates that co-cultured macrophages and fibroblasts from patients with SSc engage in reciprocal activation. However, the mechanism by which these cell types communicate and contribute to fibrosis and inflammation in SSc is unknown.

Methods

Fibroblasts were isolated from skin biopsies obtained from 7 SSc patients or 6 healthy age and gender-matched control subjects following written informed consent. Human donor-derived macrophages were cultured with exosomes isolated from control or SSc fibroblasts for an additional 48 h. Macrophages were immunophenotyped using flow cytometry, qRT-PCR and multiplex. For mutual activation studies, exosome-activated macrophages were co-cultured with SSc or healthy fibroblasts using Transwells.

Results

Macrophages activated with dermal fibroblast-derived exosomes from SSc patients upregulated surface expression of CD163, CD206, MHC Class II and CD16 and secreted increased levels of IL-6, IL-10, IL-12p40 and TNF compared with macrophages incubated with healthy control fibroblasts (n = 7, P < 0.05). Exosome-stimulated macrophages and SSc fibroblasts engaged in reciprocal activation, as production of collagen and fibronectin was significantly increased in SSc fibroblasts receiving signals from SSc exosome-stimulated macrophages (n = 7, P < 0.05).

Conclusion

In this work, we demonstrate for the first time that human SSc dermal fibroblasts mediate macrophage activation through exosomes. Our findings suggest that macrophages and fibroblasts engage in cross-talk in SSc skin, resulting in mutual activation, inflammation, and extracellular matrix (ECM) deposition. Collectively, these studies implicate macrophages and fibroblasts as cooperative mediators of fibrosis in SSc and suggest therapeutic targeting of both cell types may provide maximal benefit in ameliorating disease in SSc patients.
First-Episode Psychosis Patients Who Deteriorated in the Premorbid Period Do Not Have Higher Polygenic Risk Scores Than Others: A Cluster Analysis of EU-GEI Data
Authors: Ferraro Laura , Quattrone Diego , La Barbera Daniele , La Cascia Caterina , Morgan Craig , Kirkbride James B , Cardno Alastair G , Sham Pak , Tripoli Giada , Sideli Lucia , Seminerio Fabio , Sartorio Crocettarachele , Szoke Andrei , Tarricone Ilaria , Bernardo Miquel , Rodriguez Victoria , Stilo Simona A , Gayer-Anderson Charlotte , de Haan Lieuwe , Velthorst Eva , Jongsma Hannah , Bart Rutten B P , Richards Alexander , Arango Celso , Menezez Paulo Rossi , Lasalvia Antonio , Tosato Sarah , Tortelli Andrea , Del Ben Cristina Marta , Selten Jean-Paul , Jones Peter B , van Os Jim ,   The WP2 EU-GEI Group, Di Forti Marta , Vassos Evangelos , Murray Robin M
Published: August 2022

Cluster studies identified a subgroup of patients with psychosis whose premorbid adjustment deteriorates before the onset, which may reflect variation in genetic influence. However, other studies reported a complex relationship between distinctive patterns of cannabis use and cognitive and premorbid impairment that is worthy of consideration. We examined whether: (1) premorbid social functioning (PSF) and premorbid academic functioning (PAF) in childhood and adolescence and current intellectual quotient (IQ) define different clusters in 802 first-episode of psychosis (FEP) patients; resulting clusters vary in (2) polygenic risk scores (PRSs) for schizophrenia (SCZ_PRS), bipolar disorder (BD_PRS), major depression (MD_PRS), and IQ (IQ_PRS), and (3) patterns of cannabis use, compared to 1,263 population-based controls. Four transdiagnostic clusters emerged (BIC = 2268.5): (1) high-cognitive-functioning (n = 205), with the highest IQ (Mean = 106.1, 95% CI: 104.3, 107.9) and PAF, but low PSF. (2) Low-cognitive-functioning (n = 223), with the lowest IQ (Mean = 73.9, 95% CI: 72.2, 75.7) and PAF, but normal PSF. (3) Intermediate (n = 224) (Mean_IQ = 80.8, 95% CI: 79.1, 82.5) with low-improving PAF and PSF. 4) Deteriorating (n = 150) (Mean_IQ = 80.6, 95% CI: 78.5, 82.7), with normal-deteriorating PAF and PSF. The PRSs explained 7.9% of between-group membership. FEP had higher SCZ_PRS than controls [F(4,1319) = 20.4, P < .001]. Among the clusters, the deteriorating group had lower SCZ_PRS and was likelier to have used high-potency cannabis daily. Patients with FEP clustered according to their premorbid and cognitive abilities. Pronounced premorbid deterioration was not typical of most FEP, including those more strongly predisposed to schizophrenia, but appeared in a cluster with a history of high-potency cannabis use.
Services, Jobs, and Economic Development in Africa
Authors: Baccini Leonardo , Fiorini Matteo , Hoekman Bernard , Sanfilippo Marco
Published: August 2022

This article presents data and analyzes the structure of employment in 13 African economies at the administrative unit level, with a focus on the role of services. We provide two novel pieces of evidence. First, we present a descriptive snapshot of changes in the composition of employment over time and across geographies. This reveals evidence of structural transformation toward services and service-related occupations at subnational level and provides a fine-grained overview of who works in services and where and how this has changed over time. Second, we provide correlations between services and economic development, using per capita nightlight luminosity as a proxy. We document (1) a strong positive association between high skills services and economic development; (2) substantial heterogeneity across industries within services; and (3) a mediating role of market conditions and technology in the relation between services and economic development. Overall, our work highlights an important role of services activities for employment, skills, and economic development in Africa.
Diversity and Pathogenicity of Fungi Associated with Fruit Rot of Winter Jujube in Shandong Province, China
Authors: Xu Xi , Guo Lifeng , Yang Chunbo , Teng Haolin , Shen Guijin , Wang Shuo , Zhao Junlei , Yang Xilang , Zhang Li , Wang Xiangjing , Zhao Junwei , Xiang Wensheng
Published: August 2022

Winter jujube originated from China and had an extremely high nutritional value. In 2021, symptomatic winter jujube fruits were collected from eight locations in Zhanhua District of Binzhou City, Shandong Province. In total, 108 fungal isolates were obtained and grouped into 11 species based on morphological characteristics and multilocus phylogenetic analysis, including Nothophoma quercina (43.52%), Fusarium lateritium (20.37%), Alternaria alternata (12.03%), F. proliferatum (7.41%), F. graminearum (4.63%), Botryosphaeria dothidea (3.70%), Fusarium sp. (2.78%), A. tenuissima (2.78%), Diaporthe eres (1.85%), Nigrospora oryzae (0.93%), and Cercospora nicotianae (0.93%). All fungal isolates obtained in this study showed aggressiveness on detached winter jujube fruits except N. oryzae and C. nicotianae isolates, of which F. proliferatum was the most virulent, while A. alternata isolates, which have been considered the major pathogen of winter jujube fruit rot, showed a relatively low-level virulence in this study. Furthermore, D. eres, F. graminearum, F. lateritium, and an unclassified Fusarium species were first reported as causal agents of winter jujube fruit rot. The typical symptoms of winter jujube fruit rot observed in this study could be distinguished into two types. N. quercina, A. alternata, A. tenuissima, Fusarium sp., D. nobilis, and F. lateritium isolates caused reddish brown to dark gray lesions on the peel, while B. dothidea, F. graminearum, and F. proliferatum isolates caused peel and pulp decay, resulting in red to reddish brown and water-soaked lesions. In addition, haplotype analysis of N. quercina isolates obtained in this study and validly published articles showed that there were 11 haplotypes worldwide; the isolates obtained in the current study were grouped into three haplotypes (Hap 1, Hap 2, and Hap 11), and two of them (Hap 2 and Hap 11) were confirmed as new haplotypes.
Ambroxol, a mucolytic agent, boostsHO‐1, suppressesNF‐κB, and decreases the susceptibility of the inflamed rat colon to apoptosis: A new treatment option for treating ulcerative colitis
Authors: Cavalu Simona , Sharaf Hossam , Saber Sameh , Youssef Mahmoud E. , Abdelhamid Amir Mohamed , Mourad Ahmed A. E. , Ibrahim Samar , Allam Shady , Elgharabawy Rehab Mohamed , El‐Ahwany Eman , Amin Noha A. , Shata Ahmed , Eldegla Mai , Atef Marina , Aboraya Maii , Mohamed Mayar , Anz Niera , Elmotelb Dina Abd , Gabr Fayrouz , Elzablawy Dalia , Hamada Menna , Yehia Amr , Osama Dalia , Mohammed Osama A.
Published: August 2022

Ulcerative colitis (UC) is a chronic inflammatory bowel disease of unknown etiology that increases the risk of developing colorectal cancer and imposes a lifelong healthcare burden on millions of patients worldwide. Current treatment strategies are associated with significant risks and have been shown to be fairly effective. Hence, discovering new therapies that have better efficacy and safety profiles than currently exploited therapeutic strategies is challenging. It has been well delineated that NF‐κB/Nrf2 crosstalk is a chief player in the interplay between oxidative stress and inflammation. Ambroxol hydrochloride, a mucolytic agent, has shown antioxidant and anti‐inflammatory activity in humans and animals and has not yet been examined for the management of UC. Therefore, our approach was to investigate whether ambroxol could be effective to combat UC using the common acetic acid rat model. Interestingly, a high dose of oral ambroxol (200 mg/kg/day) reasonably improved the microscopic and macroscopic features of the injured colon. This was linked to low disease activity and a reduction in the colonic weight/length ratio. In the context of that, ambroxol boosted Nrf2 activity and upregulated HO‐1 and catalase to augment the antioxidant defense against oxidative damage. Besides, ambroxol inactivated NF‐κB signaling and its consequent target pro‐inflammatory mediators, IL‐6 and TNF‐α. In contrast, IL‐10 is upregulated. Consistent with these results, myeloperoxidase activity is suppressed. Moreover, ambroxol decreased the susceptibility of the injured colon to apoptosis. To conclude, our findings highlight the potential application of ambroxol to modify the progression of UC by its anti‐inflammatory, antioxidant, and antiapoptotic properties.
Transforming Clinical Education During a Global Pandemic: Building New Approaches to Clinical Education
Authors: Rembish Melissa , Snyder Marianne , Sarage Dawn , Spadafora Nancy , Alexander Ivy M.
Published: August 2022

The sudden and severe impact of COVID-19 on nursing education brought about many challenges and opportunities. The Community of Inquiry model and Online Collaborative Learning theory provided a guiding framework for preparing faculty to develop curriculum and deliver online/virtual synchronous clinical and laboratory experiences for junior and senior nursing students, overcome identified challenges, and evaluate the experiences. The swift change to virtual clinical education required careful reflection about how we teach, evaluate learning, and move forward with precision while establishing new best practices.
Facilitating Residents’ Understanding of Electronic Health Record Report Card Data Using Faculty Feedback and Coaching
Authors: Sebok-Syer Stefanie S. , Shaw Jennifer M. , Sedran Robert , Shepherd Lisa , McConnell Allison , Dukelow Adam M. , Syer Mark D. , Lingard Lorelei
Published: August 2022

Purpose

Feedback continues to present a challenge for competency-based medical education. Clear, consistent, and credible feedback is vital to supporting one’s ongoing development, yet it can be difficult to gather clinical performance data about residents. This study sought to determine whether providing residents with electronic health record (EHR)-based report cards, as well as an opportunity to discuss these data with faculty trained using the R2C2 model, can help residents understand and interpret their clinical performance metrics.

Method

Using action research methodology, the author team collected EHR data from July 2017 to February 2020, for all residents (n = 21) in one 5-year Emergency Medicine program and created personalized report cards for each resident. During October 6–17, 2020, 8 out of 17 eligible residents agreed to have their feedback conversations recorded and participate in a subsequent interview with a nonphysician member of the research team. Data were analyzed using thematic analysis, and the authors used inductive analysis to identify themes in the data.

Results

In analyzing both the feedback conversations as well as the individual interviews with faculty and residents, the authors identified 2 main themes: (1) Reactions and responses to receiving personalized EHR data and (2) The value of EHR data for assessment and feedback purposes. All participants believed that EHR data metrics are useful for prompting self-reflection, and many pointed to their utility in providing suggestions for actionable changes in their clinical practice. For faculty, having a tool through which underperforming residents can be shown “objective” data about their clinical performance helps underscore the need for improvement, particularly when residents are resistant.

Conclusions

The EHR is a valuable source of educational data, and this study demonstrates one of the many thoughtful ways it can be used for assessment and feedback purposes.
Optimizing Clinical Reasoning Assessments With Analytic and Holistic Ratings: Examining the Validity, Reliability, and Cost of a Simplified Patient Note Scoring Procedure
Authors: Cheung Jeffrey J.H. , Park Yoon Soo , Aufderheide Kassidee , Holden Jaime , Yudkowsky Rachel
Published: August 2022

Purpose

Post-standardized patient (SP) encounter patient notes used to assess students’ clinical reasoning represent a significant time burden for faculty who traditionally score them. To reduce this burden, the authors previously reported a complex faculty-developed scoring method to assess patient notes rated by nonclinicians. The current study explored whether a simplified scoring procedure for nonclinician raters could further optimize patient note assessments by reducing time, cost, and creating additional opportunities for formative feedback.

Method

Ten nonclinician raters scored patient notes of 141 students across 5 SP cases by identifying case-specific patient note checklist items. The authors identified the bottom quintile of students using the proportion of correct items identified in the note (percent-scores) and case-specific faculty-generated scoring formulas (formula-scores). Five faculty raters scored a subset of notes from low, borderline, and high-performing students (n = 30 students) using a global rating scale. The authors performed analyses to gather validity evidence for percent-scores (i.e., relationship to other variables), investigate its reliability (i.e., generalizability study), and evaluate its costs (i.e., faculty time).

Results

Nonclinician percent- and formula-scores were highly correlated (r = .88) and identified similar lists of low-performing students. Both methods demonstrated good agreement for pass-fail determinations with each other (Kappa = .68) and with faculty global ratings (Kappapercent =.61; Kappaformula =.66). The G-coefficient of percent-scores was .52, with 38% of variability attributed to checklist items nested in cases. Using percent-scores saved an estimated $746 per SP case (including 6 hours of faculty time) in development costs over formula-scores.

Conclusions

Nonclinician percent-scores reliably identified low-performing students without the need for complex faculty-developed scoring formulas. Combining nonclinician analytic and faculty holistic ratings can reduce the time and cost of patient note scoring and afford faculty more time to coach at-risk students and provide targeted assessment input for high-stakes summative exams.
Racial/Ethnic Disparities in Clerkship Grading: Perspectives of Students and Teachers
Authors: Hanson Janice L. , Pérez Maria , Mason Hyacinth R.C. , Aagaard Eva M. , Jeffe Donna B. , Teherani Arianne , Colson Eve R.
Published: August 2022

Purpose

Racial/ethnic disparities exist in clinical clerkship grading, yet little is known about medical student and faculty perspectives on why these disparities occur. This study explored what happens during clerkships that might explain grading disparities.

Method

Medical students and clerkship teachers at 3 U.S. medical schools completed a demographic survey and semistructured interview. The constant comparative method was used to analyze transcripts by inductively developing codes; grouping codes in categories; and refining codes, descriptions, and group assignments to identify themes. Interpretations of and relationships among themes were iteratively discussed to develop a grounded theory.

Results

Fifty-nine participants (29 medical students, 30 teachers [28 clinical faculty, 2 residents]) were interviewed in 2020. The Social Milieu of Medical Education (relationships, fit, opportunities, and judgments in the clinical-learning setting) was the organizing theme, influenced by 5 additional themes: Societal Influence (experiences in society), Students’ Characteristics and Background (personal characteristics and experiences outside medical school), Assessment Processes (collection of student performance data and how data inform grades), Learning Environment (resources available and messaging within the clinical setting), and Students’ Interactions and Reactions (interactions with and reactions to peers and teachers). The grounded theory highlights complex, multilayered aspects of how the social milieu of medical education is shaped by and shapes students’ experiences, relationships, and clerkship assessments and promotes clerkship-grading disparities.

Conclusions

Mitigating clerkship-grading disparities will require intervening on interrelated, contextual factors to provide equitable opportunities for students from diverse backgrounds and with varying styles of engagement in clinical-learning settings, along with attending to modifying assessment processes.
Medical Education Systems in China: Development, Status, and Evaluation
Authors: Liu Xihan , Feng Jie , Liu Chenmian , Chu Ran , Lv Ming , Zhong Ning , Tang Yuchun , Li Li , Song Kun
Published: August 2022

Since 1949, China has made many changes to develop its medical education system and now has a complex array of medical degrees. The current system comprises a 3-year junior college medical program, 5-year medical bachelor’s degree program, “5 + 3” medical master’s degree program, and 8-year medical doctoral degree program; these programs each provide a different path to earning a medical degree. The advantages and drawbacks of such complexity are open to discussion. Since the government set a strategic goal of “Healthy China” in 2019, it has sought to increase the training capacity of its medical education system to establish a high-quality health service system. This article reviews medical education reform in China, discusses the current medical education system, and presents evaluations of medical education programs based on assessments by 1,025 participants (medical students and doctors) recruited from 31 provinces of China. These assessments were compiled via a multicenter self-reported questionnaire administered July 1 to 5, 2021. Participants were training for a medical degree or practicing doctors trained in the 5-year program, “5 + 3” program, 8-year program, or “4 + 4” program. The authors assessed the medical education system to which each of the participants belong and their career stage and career satisfaction, and they requested that participants name the 3 most promising programs. The 8-year program ranked first in work satisfaction (7.92/10), education program satisfaction (7.78/10), and potential (1.91/2). Scores of the 5-year program and “5 + 3” programs were 7.25 and 7.17 for system satisfaction, respectively, and the “4 + 4” program (7.00/10) ranked the next highest. The innovations that have occurred in the Chinese medical education system have offered opportunities to meet the needs of more patients, but the lack of consistency has also posed challenges. Currently, Chinese medical education is becoming more uniform and standardized.
Abortion as Essential Health Care and the Critical Role Your Practice Can Play in Protecting Abortion Access
Authors: Fay Kathryn E. , Diouf Khady , Butler Sharlay K. , Onwuzurike Chiamaka , Wilkinson Barbara E. , Johnson Natasha R. , Schantz-Dunn Julianna , Bartz Deborah
Published: August 2022

Few obstetrician–gynecologists (ob-gyns) provide abortion care, resulting in abortion’s being separated from other reproductive health care. This segregation of services disrupts the ob-gyn patient–clinician relationship, generates needless costs, delays access to abortion care, and contributes to stigma. General ob-gyns have both the skills and the knowledge to incorporate abortion into their clinical practices. In this way, they can actively contribute to the protection of abortion access now, with the loss of federal protection for abortion under Roe v Wade. For those who live where abortion remains legal, now is the time to start providing abortions and enhancing your abortion-referral process. For all, regardless of state legislation, ob-gyns must be leaders in advocacy by facilitating abortion care—across state lines, using telehealth, or with self-managed abortion—and avoiding any contribution to the criminalization of those who seek or obtain essential abortion care. Our patients deserve a specialty-wide concerted effort to deliver comprehensive reproductive health care to the fullest extent.
Medial Head of the Gastrocnemius Tenotomy Through a Posteromedial Approach Significantly Improves Surgical Accessibility of the Tibial Plateau
Authors: Cardon Jeffrey J. , DeKeyser Graham J. , Peterson Andrew C. , Higgins Thomas F. , Lenz Amy L. , Haller Justin M.
Published: August 2022

Objectives:

Compare accessible area of the posterior tibial plateau through a modified posteromedial (PM) approach before and after tenotomy of the medial head gastrocnemius. We report the outcomes of 8 patients who underwent gastrocnemius tenotomy during PM approach.

Methods:

A modified PM approach was performed on 10 cadaveric legs, and the surgically accessible area was outlined. Next, a medial head gastrocnemius mid substance tenotomy was completed, and the accessible area was again outlined. Tibia specimens were imaged in a micro-CT scanner to measure accessible surface area and linear distance along the joint line. In addition, 8 patients who underwent tenotomy for tibial plateau fracture had outcomes recorded.

Results:

The modified PM approach with tenotomy provided significantly more access to the posterior plateau than without tenotomy. The modified PM approach before tenotomy allowed access to 1774 mm2 (SD = 274) of the posterior plateau surface and 2350 mm2 (SD = 421, P < 0.0001) with tenotomy. A linear distance of 38 mm (SD = 7) and 57 mm (SD = 7, P < 0.00001) was achieved before and after tenotomy, respectively. In the clinical series, the average knee arc of motion was 116 degrees (95–135).

Conclusions:

The modified PM approach with medial head gastrocnemius tenotomy significantly improves surgical access to the posterior plateau. Patients who received tenotomy have acceptable functional outcomes. This cadaveric study provides an alternative approach for treatment of posterolateral tibial plateau fractures which may mitigate damage to neurovascular structures.
LINC00657 regulate colorectal carcinoma invasion and migration by enhancing heparanase expression through recruiting SMAD family member 2
Authors: Gong Shuangxi , Yang Fengshuai , Song Yuliang , Wang Xinyu , Yang Yunquan
Published: August 2022

Long noncoding RNAs are master regulators of several cancer phenotypes, such as cell growth, apoptosis, and motility. This study is designed to resolve the relevance of LINC00657 with tumor invasion and migration and its action mechanism in colorectal carcinoma (CRC). LINC00657 and HPSE levels were first examined in cancerous tissues from CRC patients and CRC cells. Then functional experiments were conducted to evaluate the abilities of HCT116 and SW620 cells to proliferate, migrate, and invade when LINC00657 or HPSE was knocked down, or LINC00657 knockdown and SMAD2 overexpression were simultaneously introduced. Snail and E-cadherin levels in the CRC cells were evaluated. Next, the binding between LINC00657 and SMAD2 or between SMAD2 and HPSE was determined. LINC00657-silencing HCT116 cells were inoculated into nude mice, and the tumorigenesis and the levels of Snail and E-cadherin were evaluated. LINC00657 and HPSE were increasingly expressed in CRC. Knockdown of LINC00657 or HPSE inhibited the malignant properties of CRC cells, decreased Snail expression, and strengthened E-cadherin level. LINC00657 and HPSE could both bind to SMAD2. SMAD2 overexpression counteracted the inhibiting effect of LINC00657 silencing on HPSE expression and the growth and invasion of CRC cells. In vivo experiments further verified the suppression of LINC00657 knockdown on tumor growth and metastasis. LINC00657 recruits SMAD2 to HPSE promoter region to elevate HPSE transcription, thus accelerating CRC invasion and migration.
LncRNA ST8SIA6-AS1 facilitates hepatocellular carcinoma progression by governing miR-651-5p/TM4SF4 axis
Authors: Mou Yanjie , Ding Xiaoming
Published: August 2022

The oncogenic role of ST8SIA6-AS1 in different cancers was reported, including hepatocellular carcinoma (HCC). However, the underlying mechanism has not been completely explored. Real time quantitative PCR analysis was conducted to assess the ST8SIA6-AS1, miR-651-5p and TM4SF4 expression in HCC tissues and cells. Cell counting kit-8 and wound-healing migration assays were adopted to evaluate the HCC cell proliferation and migration, respectively. The expression of apoptosis-related proteins (Bax and Bcl-2) in human colorectal cancer cells (HCC) was determined by western blotting. In addition to bioinformatics analysis, RNA immunoprecipitation studies and luciferase reporter assays were undertaken to investigate the direct target relationship among ST8SIA6-AS1 and miR-651-5p or TM4SF4. Highly expressed ST8SIA6-AS1 and TM4SF4 as well as poorly expressed miR-651-5p were detected in HCC tissues and cells. Clinically, miR-651-5p expression in HCC tissues is negatively correlated with ST8SIA6-AS1 or TM4SF4. Cell functional assays demonstrated that ST8SIA6-AS1 silencing resulted in weakened proliferative and migratory capacities in HCC cells in addition to increase Bax expression and reduced Bcl-2 expression. ST8SIA6-AS1 exhibited its oncogenic function by sponging tumor suppressor miR-651-5p, and the anti-oncogenic of miR-651-5p was offset by its TM4SF4. The manipulation of ST8SIA6-AS1/miR-651-5p/TM4SF4 axis-mediated oncogenicity in HCC might shed new light on HCC diagnosis and therapy.
GATA3 positively regulates PAR1 to facilitate in vitro disease progression and decrease cisplatin sensitivity in neuroblastoma via inhibiting the hippo pathway
Authors: Wang Jing , Dai Wang , Zhang Ming
Published: August 2022

GATA binding protein 3 (GATA3) is reported to critically involved in the pathogenesis of neuroblastoma (NB). This study investigated the specific role and mechanism of GATA3 in NB progression. JASPAR was utilized to predict GATA3’s downstream targets, whose binding relation with GATA3 was inspected by a dual-luciferase reporter assay. NB cell lines underwent transfection of GATA3/protease-activated receptor 1 (PAR1) overexpression plasmids or shGATA3, followed by cisplatin treatment. NB cell sensitivity to cisplatin, viability, migration, invasion, cell cycle progression and apoptosis were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, wound healing assay, transwell assay and flow cytometry, respectively. Expressions of GATA3, PAR1, epithelial-mesenchymal transition-related molecules (N-Cadherin and Vimentin), hippo pathway-related molecules (mammalian Ste20-like kinase (Mst)1, Mst2, Mps one binding (Mob) 1, phosphorylated (p)-Mob1, Yes-associated protein (YAP) and p-YAP) in NB tissues and cell lines were assessed by western blot or qRT-PCR. GATA3 expression was increased in NB tissues and cells. GATA3 overexpression increased NB cell viability, promoted migration, invasion, and cell cycle progression, increased the expressions of N-Cadherin, Vimentin and YAP, decreased the expressions of Mst1, Mst2, Mob1, p-Mob1, p-YAP and the ratio of p-YAP to YAP, and attenuated cisplatin-induce cell apoptosis, which GATA3 knockdown induced the opposite effect. GATA3 directly targeted PAR1, whose overexpression increased NB cell viability, inhibited the hippo pathway, and attenuated cisplatin-induce cell apoptosis, and reversed GATA3 knockdown-induced effect on these aspects. GATA3 positively regulates PAR1 to facilitate in-vitro disease progression and decrease cisplatin sensitivity in NB via inhibiting the hippo pathway.
A novel intergenic region ALK fusion is targetable by alectinib in a non-small cell lung cancer patient with brain metastasis
Authors: Cheng Wanwan , Qian Chunfa , Zhang Haitao , Meng Qi , Yin Jiani C. , Fang Shencun
Published: August 2022

Anaplastic lymphoma kinase (ALK) rearrangement defines a unique nonsmall cell lung cancer (NSCLC) molecular subtype, of which the patients could potentially benefit from anti-ALK therapies. So far, the outcomes of the canonical echinoderm microtubule-associated protein-like (EML-ALK) patients subjected to ALK inhibitors are well established. However, given the increasing complexity of ALK fusion partners, as detected by high-throughput sequencing, the responses of those with rare ALK fusion events remain to be explored. Here, we report a lung adenocarcinoma patient with brain metastasis harboring an ARHGAP5 downstream intergenic region ALK fusion, as detected by using DNA-based next-generation sequencing, who experienced a partial response to alectinib treatment. While whole- transcriptome RNA sequencing (RNA-seq) failed to identify potential ALK fusion transcripts, subsequent targeted deep RNA-seq revealed the expression of EML4-ALK transcripts in the tumor tissue. Given the increasing application of the ALK-tyrosine kinase inhibitors (TKIs), it is extremely crucial to define the patients who could be suitable for this treatment in clinic. The present case has provided supporting evidence that noncanonical ALK rearrangements on the genomic level are often functionally relevant and targetable by ALK-TKI, particularly in cases with sub-optimal quantity and quality for RNA validation.