bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expansion microscopy resolves the thylakoid structure of spinach
Authors: Bos Peter R , Berentsen Jarne , Wientjes Emilie 
Published: October 2023

Abstract
The light-harvesting reactions of photosynthesis take place on the thylakoid membrane inside chloroplasts. The thylakoid membrane is folded into appressed membranes, the grana, and nonappressed membranes that interconnect the grana, the stroma lamellae. This folding is essential for the correct functioning of photosynthesis. Electron microscopy and atomic force microscopy are commonly used to study the thylakoid membrane, but these techniques have limitations in visualizing a complete chloroplast and its organization. To overcome this limitation, we applied expansion microscopy (ExM) on isolated chloroplasts. ExM is a technique that involves physically expanding a sample in a swellable hydrogel to enhance the spatial resolution of fluorescence microscopy. Using all-protein staining, we visualized the 3D structure of spinach (Spinacia oleracea) thylakoids in detail. We were able to resolve stroma lamellae that were 60 nm apart and observe their helical wrapping around the grana. Furthermore, we accurately measured the dimensions of grana from top views of chloroplasts, which allow for precise determination of the granum diameter. Our results demonstrate that ExM is a fast and reliable technique for studying thylakoid organization in great detail.
Fluorescein staining of chloroplast starch granules in living plants
Authors: Ichikawa Shintaro , Sakata Momoko , Oba Toru , Kodama Yutaka 
Published: October 2023

Abstract
Chloroplast starch granules (cpSGs) store energy harvested through photosynthesis in plants, and cpSG dynamics have important roles in plant energy metabolism and stress responses. To date, cpSGs have been visualized using several methods, such as iodine staining; however, no method can be used to specifically visualize cpSGs in living cells from various plant species. Here, we report a simple method to visualize cpSGs in living plant cells in various species by staining with fluorescein, a commonly used fluorescent dye. We show that fluorescein is taken up into chloroplasts and interacts with cpSGs similarly to iodine. Fluorescein also interacts with refined starch in vitro. Using a fluorescein derivative for ultrabright cpSG imaging, we produced high-quality 3D reconstructions of cpSGs and evaluated their accumulation in multiple plant species. As fluorescein is well known and readily purchasable, our fluorescein-based staining method should contribute to all research regarding starch.
Off to a slow start: which workplace policies can limit gender pay gaps across firm tenure?
Authors: Kronberg Anne-Kathrin , Gerlach Anna 
Published: October 2023

Abstract
Much of the gender pay gap is generated within workplaces, making it paramount to understand which workplace policies effectively address gaps. Our article asks when policies limit gender pay gaps across employee tenure to identify potential temporal weak points. We analyze a representative panel of 10,000 establishments with over 850,000 employees using the 2005–19 waves of German-linked employer–employee data (LIAB). Two key findings emerge. First, a temporal perspective on workplace policies reveals that no policy under study—formalization, identity-based career programs, and child care assistance—reduces gender pay gaps at hire. Instead, policies only address additional disparities that accumulate after hire. Second, only identity-based career programs narrow gender disparities for all women. In contrast, seemingly gender-neutral formalization is insufficient, while providing employer-sponsored child care has mixed effects depending on employees’ education. We conclude by discussing the implications of these findings for organizational policy and future research.
Leflunomide alleviates obesity via activation of the TAK1‐AMPK pathway and induction of lipophagy
Authors: Ji Xiaoyue , Chen Junhong , You Chaoying , Sun Jing , Xu Xiulong 
Published: October 2023

AbstractLipophagy is a subset of selective autophagy that specifically degrades lipid droplets and plays an important role in obesity. Leflunomide treatment in rheumatoid arthritis (RA) patients has been associated with weight loss and decreased blood glucose levels, which cannot be attributed to its known side effects. Our prior studies showed that A77 1726, the active metabolite of leflunomide, acts as an inhibitor of S6K1 to sensitize the insulin receptor and control hyperglycemia. Whether the anti‐obesity effect of leflunomide is mediated by targeting S6K1 and its underlying mechanisms remain unclear. Here, we report that A77 1726 induced LC3 lipidation and increased the formation of autophagosomes and lipoautolysosomes in 3T3‐L1 adipocytes by activating TGF‐β‐activated kinase 1 (TAK1), AMP‐activated kinase (AMPK), and Unc‐51 like autophagy‐activated kinase 1 (ULK1). A77 1726 reduced the content of lipid droplets in 3T3‐L1 adipocytes, which was blocked by bafilomycin or by beclin‐1 knockdown. Similar observations were made in murine adipocytes differentiated from S6K1−/− embryonic fibroblasts (MEFs). Leflunomide treatment restricted bodyweight gains in ob/ob mice and reduced the visceral fat deposit and the size of adipocytes. Leflunomide treatment induced autophagy in adipose and liver tissues and reduced hepatic lipid contents. Consistently, S6K1 knockout increased the levels of LC3 lipidation in the liver, muscle, and fat of S6K−/− mice. Leflunomide treatment and S6K1 deficiency both induced TAK1, AMPK, and ULK1 phosphorylation in these tissues. These observations collectively suggest that leflunomide controls obesity in part by activating AMPK and inducing lipophagy. Our study provides insights into the mechanisms of leflunomide‐mediated anti‐obesity activity.
The Psychological and Biological Benefits of Mind-Body Therapy Interventions for Informal Caregivers of Individuals With Cancer: A Systematic Review
Authors: Roderick Quinn , Kuil Kristina , Kunkler Elizabeth , Mondi Nina , Rhode Aminah , Moriello Gabriele 
Published: October 2023

Background:
Caring for an individual with cancer can be physically and emotionally draining and finding an effective intervention that supports caregivers is needed. The purpose of this systematic review was to investigate the psychological and biological effects of mind-body therapies for informal caregivers of individuals with cancer.

Methods:
A literature search was completed using 6 databases (PubMed, CINAHL, Cochrane, PsycINFO, PEDro, and OTSeekers) and 4 gray literature databases (MedNar, NY Academy of Medicine Grey Literature Report, WorldCat Dissertations and Theses, and Proquest Psychology). Studies were included if they were randomized controlled trials published after 2010; participants were informal caregivers of individuals with cancer; and the main intervention was yoga, meditation, progressive relaxation therapy and guided imagery, mindfulness therapy, tai chi, breathing exercises, qigong, or visual imagery. The researchers completed title, abstract, and full-text screens using the selection criteria. Data were synthesized descriptively according to the different types of interventions. The Physiotherapy Evidence Database (PEDro) scale was used to critically appraise the final articles.

Results:
There were a total of 11 articles included in this systematic review. There is evidence that certain mind-body therapies can have a positive effect on well-being, anxiety, and fatigue but not mindfulness or stress. One study found significant improvements in biological markers.

Conclusion:
Medical professionals should consider recommending mind-body therapies, especially to informal caregivers who present with anxiety, fatigue, and impairments in well-being. The major limitation in this review is based mostly on fair quality evidence.
Incorporating Data from Multiple Endpoints in the Analysis of Clinical Trials: Example from RSV Vaccines
Authors: Prunas Ottavia , Willemsen Joukje E. , Bont Louis , Pitzer Virginia E. , Warren Joshua L. , Weinberger Daniel M. 
Published: October 2023

Background:
To meet regulatory approval, interventions must demonstrate efficacy against a primary outcome in randomized clinical trials. However, when there are multiple clinically relevant outcomes, selecting a single primary outcome is challenging. Incorporating data from multiple outcomes may increase statistical power in clinical trials. We examined methods for analyzing data on multiple endpoints, inspired by real-world trials of interventions against respiratory syncytial virus (RSV).

Method:
We developed a novel permutation test representing a weighted average of individual outcome test statistics (wavP) to evaluate intervention efficacy in a multiple endpoint analysis. We compared the power and type I error rate of this approach to the Bonferroni correction (bonfT) and the minP permutation test. We evaluated the different approaches using simulated data from three hypothetical trials varying the intervention efficacy, correlation, and incidence of the outcomes, and data from a real-world RSV clinical trial.

Results:
When the vaccine efficacy against different outcomes was similar, wavP yielded higher power than bonfT and minP; in some scenarios the improvement in power was substantial. In settings where vaccine efficacy was notably larger against one endpoint compared with the others, all three methods had similar power. We developed an R package, PERmutation basEd ANalysis of mulTiple Endpoints (PERMEATE), to guide the selection of the most appropriate method for analyzing multiple endpoints in clinical trials.

Conclusions:
Analyzing multiple endpoints using a weighted permutation method can increase power, whereas controlling the type I error rate compared with established methods under conditions mirroring real-world RSV clinical trials.
Do Unpaid Children’s Hospital Account Balances Correlate with Family Income or Insurance Type?
Authors: Yau Alice , Lentskevich Marina A. , Yau Irene , Reddy Narainsai K. , Ahmed Kaleem S. , Gosain Arun K. 
Published: October 2023

Background:Current understanding of medical debt among various income ranges and insurance carriers is limited. We analyzed median household incomes, insurance carriers, and medical debt of plastic surgery patients at a major metropolitan children’s hospital.Methods:A retrospective chart review for zip codes, insurance carriers, and account balances was conducted for 2018–2021. All patients were seen by members of the Division of Pediatric Plastic Surgery at Ann and Robert H. Lurie Children’s Hospital of Chicago. Blue Cross was reported separately among other commercial insurance carriers by the hospital’s business analytics department. Median household income by zip code was obtained. IBM SPSS Statistics was used to perform chi-squared tests to study the distribution of unpaid account balances by income ranges and insurance carriers.Results:Of the 6877 patients, 630 had unpaid account balances. Significant differences in unpaid account balances existed among twelve insurance classes (P< 0.001). There were significant differences among unpaid account balances when further examined by median household income ranges for Blue Cross (P< 0.001) and other commercial insurance carriers (P< 0.001).Conclusions:Although patients with insurance policies requiring higher out-of-pocket costs (ie, Blue Cross and other commercial insurance carriers) are generally characterized by higher household incomes, these patients were found to have higher unpaid account balances than patients with public insurance policies. This suggests that income alone is not predictive of unpaid medical debt and provides greater appreciation of lower income families who may make a more consistent effort in repaying their medical debt.
A Novel Doppler Flowmetry Shaft for Postoperative Monitoring after Head and Neck Reconstructive Surgery
Authors: Onoda Satoshi , Satake Toshihiko , Katsuragi Ryohei , Kobayashi Kohta , Tsukura Kahori , Tachibana Gaku 
Published: October 2023

Summary:
Doppler flowmetry is one of the most popular methods of monitoring Doppler signals during reconstructive surgery of the body surface. However, because of the thick and straight structure of the shaft, it is difficult to perform in areas with limited space, such as the oral cavity. We used a new type of Doppler flowmetry shaft to postoperatively monitor the flap in the oral cavity. Compared with conventional Doppler flowmetry, the new type uses a thinner metal probe shaft that can easily be inserted in narrow and limited spaces, such as the oral cavity. Additionally, the tip of the metal probe is gently bent, thereby allowing the Doppler tip to be placed perpendicular to the surface of the skin flap. We used this new type of Doppler flowmetry shaft for 30 patients after head and neck reconstruction using free flap transfer because Doppler signals were difficult to hear using conventional Doppler flowmetry. For all 30 patients, the new Doppler flowmetry shaft was able to monitor free flaps. Vascular thrombosis or vascular spasm occurred in three patients; two patients had inadequate arterial flow caused by vasospasm and arterial thrombus, and one patient had a venous thrombus. These three patients required re-exploration, and all flaps survived. This new type of Doppler flowmetry is simple and noninvasive. Furthermore, it can easily be performed by nonphysician medical personnel, and is useful for monitoring patients after head and neck reconstructive surgery.
Vascularized Toe Proximal Interphalangeal Joint Transfer in Posttraumatic Fingers: Analysis of Prognostic Factors for Suboptimal Outcomes
Authors: Huang Chi-Wei , Lin Ming-Huei , Lee Che-Hsiung , Chen Shih-Heng , Hsu Chung-Cheng , Lin Cheng-Hung , Huang Ren-Wen , Lin Yu-Te 
Published: October 2023

Background:
Posttraumatic finger osteoarthritis of the proximal interphalangeal joint (PIPJ) is a difficult problem. Over the past decade, we have reported several methods for improving the outcomes of vascularized toe joint transfer (VJT). In this study, we focused on determining poor prognostic factors which lead to a suboptimal outcome.

Methods:
A consecutive series of patients with posttraumatic osteoarthritis of the PIPJ who received VJT between January 2008 and January 2021 were enrolled in this study. The senior surgeon (Y.-T.L.) performed the surgery in all cases. In this retrospective study, we reexamine the initial trauma-related soft tissue and bony structure injuries of the recipient finger, to assess the baseline tissue quality before VJT. The injuries were classified into five major categories according to their anatomic region. The functional outcome parameters (including range of motion, percentage of use, and extensor lag of the transferred PIPJ) were collected. Univariate and multivariate linear regression analyses were performed using the generalized estimated equation model to identify the correlation between the injury category involved and functional outcome.

Results:
A total of 59 digits were enrolled. Our results revealed that the fingers with previous vascular injury that received revascularization procedures had relatively suboptimal functional outcomes. These fingers had a significantly lower percentage of use both before (β = −0.222, P = 0.006) and after (β = −0.177, P = 0.006) receiving secondary procedures to improve functional outcome.

Conclusions:
Patients with prior revascularization surgery were associated with a poor functional outcome after VJT.
Surgical or Nonsurgical Facial Rejuvenation: The Patients’ Choice
Authors: Corduff Niamh 
Published: October 2023

Background:
Patients seeking a pan-facial rejuvenation may not receive appropriate medical advice on surgical versus nonsurgical treatments, resulting in those who are best-suited for surgery receiving nonsurgical options, and vice versa. Patients who requested total-face, nonsurgical revitalization were surveyed to understand the factors influencing this decision-making.

Methods:
Patients consulting for total facial rejuvenation were surveyed on their reasons for seeking nonsurgical versus surgical interventions by an investigator with 30 years of plastic surgery experience, practicing in a nonsurgical clinic alongside a dermatologist and aesthetic physician.

Results:
Of the 92 patients surveyed, 78% completed the survey, 47% of whom had considered facelift surgery, and 14% of whom proceeded to inquiries or consultations with a plastic surgeon about facelift surgery. All respondents were women, and age was not an exclusion factor. Forty-four percent would still consider facelifts in later life. Among the most common reasons for choosing nonsurgical approaches were a desire for natural and subtle results, cost, having flexibility in treatment choice, concerns about surgical and anesthesia risks, downtime, the ability to distribute treatments over time, and scarring.

Conclusions:
Many patients book nonsurgical treatments without prior professional advice, and consider these as a temporary solution, but may desire surgery later in life. Before conducting nonsurgical treatments, doctors should identify this group and avoid interventions that may interfere with optimal surgical outcomes later. Also, some patients desire surgical outcomes when surgery is not a feasible option, and thus seek nonsurgical alternatives. For this group, realistic goals must be set before treatment to avoid disappointment.
Long-term outcome of minimally invasive staging surgery for clinical stage I endometrial cancer: A single institute experience in Taiwan
Authors: Lu Ting-Fang , Lu Chien-Hsing , Sun Lou , Liu Chi-Ku , Shih Yu-Hsiang , Hwang Sheau-Feng , Hsu Shih-Tien 
Published: October 2023

Background:
Endometrial cancer is the most common gynecological cancer in developed countries. With recent advances in equipment and knowledge, minimally invasive surgery (MIS) is widely accepted for the treatment of endometrial cancer. This study had the largest number of cases to date in Taiwan, comparing outcomes between MIS and laparotomy staging surgery using real-world data with long-term follow-up.

Methods:
We retrospectively reviewed patients with clinical stage 1 endometrial cancer from 2009 to 2020 in our institute. All patients underwent comprehensive surgical staging procedures by MIS or laparotomy. The safety, morbidity, progression-free survival (PFS), and overall survival (OS) rates of the two groups were compared. Clinical and pathologic factors were compared with Chi-square and Fisher Exact test. PFS and OS were estimated by the Kaplan-Meier method. Differences between survival curves were analyzed using the log-rank test. A p value of <0.05 was considered statistically significant. Using Cox proportional hazards models, all factors found to be significantly associated with risk of recurrence on univariate analyses were then assessed together through multivariable models, resulting in a final oncologic outcome between MIS and laparotomy.

Results:
A total of 665 cases (412 cases in MIS group and 253 cases in laparotomy group) were enrolled for data analysis. Median operation time was shorter in MIS group (244 and 265 minutes, p < 0.001). Median blood loss was also less (75 and 430 mL, p < 0.001). Median postoperative hospitalization duration was longer in the laparotomy group (2 and 7 days, p = 0.001). After adjusting presurgery risk factors, the PFS and OS were no significant difference in MIS and laparotomy groups.

Conclusion:
Using real-world data with long-term follow-up, we could confirm excellent PFS and OS in selective patients with clinical stage 1 endometrial carcinoma who received MIS, and the surgical time, hospital day, and blood loss were also less.
Physician Assistants/Associates With Doctoral Degrees: Where Are They Now?
Authors: Klein Alicia , Kayingo Gerald , Schrode Katrina M. , Soria Krista 
Published: October 2023

PurposeThe impact of holding a doctoral credential by clinically practicing physician assistants/associates (PAs) remains unknown. The purpose of this study was to investigate practice patterns and describe demographic characteristics of PAs who have terminal doctoral degrees.MethodsThis was a secondary analysis of data obtained from the 2021 Salary Survey of the American Academy of PAs. A total of 13,865 PAs responded to the survey for an estimated response rate of 13.3%. We examined the relationships between holding a doctoral degree, demographic characteristics, and work-related variables. Descriptive bivariate statistics and chi-square tests were used for data analysis.ResultsMost of the PAs (93.6%) with doctoral degrees were older than 30 years and self-identified as female (55%). There were higher proportions of non-White individuals among those with doctorates compared with general PA population. Of the PAs holding doctoral degrees, 90.4% were primarily clinicians and 9.6% were primarily educators. Physician assistants/associates with doctoral degrees were employed with their current employer longer than PAs without doctoral degrees. Most of the PAs with doctoral degrees held informal or formal leadership and had an annual salary above the median (62.3%) compared with PAs without doctoral degrees (40.0%).ConclusionThere are statistically significant differences in practice patterns and demographic factors between PAs with and without doctoral degrees. Physician assistants/associates with doctoral degrees were older, were male, and held leadership positions. Taken together, holding a doctoral credential may improve upward mobility at workplaces. Qualitative studies are warranted to further understand the motivation and impact of holding a doctoral credential among clinically practicing PAs.
Update on the genetic basis of obstructive sleep apnoea – hype or hope?
Authors: Riha Renata L. 
Published: October 2023

Purpose of review
The obstructive sleep apnoea syndrome (OSAS) is a chronic, common condition in western societies which can lead to adverse cardiometabolic effects if left untreated and is one of the commonest causes of excessive daytime somnolence.

Recent findings
The presentation of OSAS is diverse and is thought to comprise of different intermediate phenotypes and endotypes in varying proportions in each individual. Unfortunately, due to its heterogeneity and the changing definitions of the disorder by workers in the field, attempts at revealing the genetic basis of OSAS has been fraught with difficulty.

Summary
This brief review presents a short update on the achievements of the past three decades in this understudied and underfunded area of endeavour in respiratory sleep medicine. The genetic underpinnings of OSAS remain elusive.