bluuebunny/crossref_metadata_2025 huggingface dataset:
Calcium Nutrition in Fig Orchards Enhance Fruit Quality at Harvest and Storage
Authors: Souza Jackson Mirellys Azevêdo , Leonel Sarita , Leonel Magali , Garcia Emerson Loli , Ribeiro Luiza Rocha , Ferreira Rafael Bibiano , Martins Rafaelly Calsavara , de Souza Silva Marcelo , Monteiro Laís Naiara Honorato , Duarte Anita Santos
Published: January 2023

Due to the high perishability of figs, calcium stands out as an important nutrient for orchard management. This study aims to study the pre-harvest applications of calcium chloride in fig orchards. The fig trees were sprayed with 0, 0.5, 1.0, 1.5 and 2.0% CaCl2, and fruits were thereafter stored for 0, 7, 14 and 21 d. The variables analysed were the concentration of calcium in leaves and fruits, weight loss, fruit decay, pectinolytic enzyme activity, and physical and chemical attributes of the fruits; CaCl2 sprays enhanced fruit Ca2+ concentration in leaves (14.03%) and fruits (29.3%) and were effective in reducing polygalacturonase and pectin methylesterase activity, weight loss and fruit decay. Pre-treatment with 1% CaCl2 provided fruits with larger diameters, greater firmness, and higher levels of total phenolic compounds in both fruit peel and pulp. The pre-harvest application at 1% CaCl2 showed to be a promising technique in producing high quality fruits and extended storage by approximately 7 to 10 days. These findings may be useful in planning new cultural practices for fig orchards that produce high-quality fruit with desirable characteristics for growers and consumers.
Pediatric Functional Neurological Symptoms Disorder: Walking Ability and Perceived Exertion Post-Pediatric Rehabilitation
Authors: Landa Jana , Gerner Maya , Eisenstein Etzyona , Barak Sharon
Published: January 2023

Background: Adolescents with functional neurological symptoms disorder (FNSD) commonly present walking abnormalities. Walking is influenced by ‘objective’ (e.g., fitness) and ‘subjective’ (e.g., fear) components. Rate of perceived exertion (RPE) reflects the interaction between these two components. This study compared the walking ability and RPE before and after rehabilitation of adolescents with FNSD to adolescents with moderate-to-severe traumatic brain injury (TBI). Factors predicting walking and RPE were also examined. Methods: Adolescents with FNSD (n = 31) and adolescents with moderate-to-severe TBI (n = 28) aged 6 to 18 years participated in the study. Participants received a multidisciplinary rehabilitation program. Six-minute walk test (6MWT) and RPE were assessed before and after rehabilitation. Results: At pre-test, the TBI group presented lower RPE than the FNSD group (3.38 ± 2.49 and 6.25 ± 2.71, respectively). In the FNSD group, pre-test 6MWT was a significant predictor of post-test 6MWT (adjusted R2 = 0.17; p = 0.01). In the TBI group, post-test 6MWT was significantly predicted by both the pre-test 6MWT and age (adjusted R2 = 0.16; p = 0.04). Conclusions: Prior to the intervention, adolescents with FNSD perceived walking as a more difficult activity than adolescents with TBI. Post-intervention, although the intervention was effective in terms of changes in 6MWT and RPE, the ‘subjective’ component still contributed to the elevated RPE of the FNSD group.
Heartbreak and Loneliness Due to Family Separations and Limited Visiting during COVID-19: A Qualitative Study
Authors: Tsui Wai-King , Yip Ka-Huen , Yip Yuk-Chiu
Published: January 2023

The COVID-19 pandemic has greatly impacted the healthcare system. In the pediatric unit, stress, uncertainty, and many unexpected challenges for many parents were frequently reported. Research has shown that parents had less contact with their children during the pandemic due to hospital restrictions. However, it is unknown how parents perceived their experiences in a pediatric unit. This study aimed to describe the lived experiences of parents who had a child in the pediatric unit during the pandemic. A qualitative descriptive approach was used to investigate parents’ experiences of having children admitted to the pediatric unit during the pandemic in Hong Kong. Eight Chinese parents participated in the interview. Three major themes emerged: (1) parents’ pediatric ward experiences during COVID-19 were emotionally isolating and overwhelming, (2) the family and family-centered care were disrupted, and (3) interactions with pediatric providers intensified or alleviated emotional distress. Integrating the above themes of experiences of emotional distress was the main characteristic of the parents’ experiences during the pandemic. Therefore, policymakers should understand the lived experiences of parents of children diagnosed with COVID-19 and should make prompt decisions to deal with both parental concerns and safety issues.
Risk Factors for Stunting among Children under Five Years in the Province of East Nusa Tenggara (NTT), Indonesia
Authors: Suratri Made Ayu Lely , Putro Gurendro , Rachmat Basuki , Nurhayati  , Ristrini  , Pracoyo Noer Endah , Yulianto Aris , Suryatma Anton , Samsudin Mohamad , Raharni
Published: January 2023

In East Nusa Tenggara Province, Indonesia, 42.6% of children under five had stunted growth in 2018, which affects both individual and communal levels. The first step in creating effective interventions is identifying the risk factors for stunting. This study aims to pinpoint the stunting risk factors in East Nusa Tenggara Province, Indonesia, by incorporating secondary data from the 2018 Indonesia Basic Health Research (RISKESDAS). This study implemented a cross-sectional design by utilizing the data of individuals who were successfully visited during the survey. Initial data screening in East Nusa Tenggara Province based on the criteria for children aged 0–59 months and stunting showed as many as 1643. Multivariate logistic regression analysis was performed to evaluate the relationship between children’s characteristics and stunting. There was a significant relationship between age group variables for younger children (aged 12–23, 24–35, and 36–47 months), mothers with low education, and children living in rural areas with the incidence of stunting in children (p-value < 0.05). The dominant factors that caused stunting in this study were the children’s age of 24–35 months (OR = 2.08, 95% CI: 1.12–3.86), mothers with low education (OR = 1.57, 95% CI: 1.18–2.08), and children living in rural areas (OR = 1.39, 95% CI: 1.01–1.91). The highest prevalence of stunting was in the group of children aged 12–23 months (45.2%). To prevent child stunting, the government must intervene for mothers with low education and those living in rural areas. Intervention includes intensive socialization about improving nutritional status during pregnancy and practicing complementary feeding and breastfeeding habits until the child is 24 months old.
Impact of Long Working Hours on Mental Health: Evidence from China
Authors: Ma Xinxin
Published: January 2023

Although previous studies have examined the impact of long working hours on mental health in China, they have not addressed the initial value and reverse causality issues. To bridge this gap in the literature, I conducted a dynamic longitudinal analysis to investigate the association between long working hours and the risk of mental illness nationwide. Using three-wave longitudinal data from the China Family Panel Studies conducted in 2014, 2016, and 2018, I adopted dynamic regression models with lagged long working hours variables to examine their association with the risk of mental illness. The results indicate that long working hours have positive and significant (p < 0.01 or p < 0.05) associations with the risk of mental illness (OR: 1.12~1.22). The effect is more significant for women, white-collar workers, and employees in micro-firms, compared with their counterparts (i.e., men, pink- and blue-collar workers, employees of large firms, and self-employed individuals). The results provide empirical evidence of the effects of long working hours on mental health in China, confirming the need to enforce the regulations regarding standard working hours and monitor regulatory compliance by companies, as these factors are expected to improve mental health.
Antimicrobial Properties of Essential Oils Obtained from Autochthonous Aromatic Plants
Authors: Boy Francisco Ramiro , Benito María José , Córdoba María de Guía , Rodríguez Alicia , Casquete Rocío
Published: January 2023

The aim of this work was to determine the antimicrobial activity of the essential oils of six plants widely distributed in the Dehesa of Extremadura, such as Calendula officinalis, Cistus ladanifer, Cistus salviifolius, Cistus multiflorus, Lavandula stoechas, and Rosmarinus officinalis. The content of total phenolic compounds (TPC) and the antimicrobial activity of the essential oils against pathogenic and spoilage bacteria and yeasts as well as aflatoxin-producing molds were determined. A great variability was observed in the composition of the essential oils obtained from the six aromatic plants. The Cistus ladanifer essential oil had the highest content of total phenols (287.32 ppm), followed by the Cistus salviifolius essential oil; and the Rosmarinus officinalis essential oil showed the lowest amount of these compounds. The essential oils showed inhibitory effects on the tested bacteria and also yeasts, showing a maximum inhibition diameter of 11.50 mm for Salmonella choleraesuis and Kregervanrija fluxuum in the case of Cistus ladanifer and a maximum diameter of 9 mm for Bacillus cereus and 9.50 mm for Priceomyces carsonii in the case of Cistus salviifolius. The results stated that antibacterial and antiyeast activity is influenced by the concentration and the plant material used for essential oil preparation. In molds, aflatoxin production was inhibited by all the essential oils, especially the essential oils of Cistus ladanifer and Cistus salviifolius. Therefore, it can be concluded that the essential oils of native plants have significant antimicrobial properties against pathogenic and spoilage microorganisms, so they could be studied for their use in the industry as they are cheap, available, and non-toxic plants that favor the sustainability of the environment of the Dehesa of Extremeña.
A Single Session of a Digital Health Tool-Delivered Exercise Intervention May Provide Immediate Relief from Pelvic Pain in Women with Endometriosis: A Pilot Randomized Controlled Study
Authors: Lutfi Muhammad , Dalleck Lance C. , Drummond Claire , Drummond Murray , Paparella Liana , Keith Caitlin E. , Kirton Michael , Falconer Laura , Gebremichael Lemlem , Phelan Caroline , Barry Christine , Roscio Kiara , Lange Belinda , Ramos Joyce S.
Published: January 2023

Background: Endometriosis is a debilitating chronic condition that is commonly associated with chronic pelvic pain, affecting approximately 10% of women of reproductive age worldwide. The general principle of pain management in this population involves both pharmacological and surgical interventions. There is also increasing interest in the use of exercise as an alternative non-pharmacological analgesic, but adherence and accessibility to face-to-face exercise-delivery modalities are poor. This study aims to determine the immediate impact of a single session of ‘supervised’ telehealth-delivered exercise compared to ‘self-managed’ virtual reality (VR)-delivered exercise on pelvic pain associated with endometriosis. Methods: Twenty-two women experiencing pelvic pain due to endometriosis were included and randomized into three groups: (i) VR-delivered exercise group (n = 8); (ii) telehealth-delivered exercise group (n = 8); and (iii) control group (n = 6). The visual analogue scale (VAS) was used to assess the severity of pelvic pain. Results: There was no statistically significant between-group difference (p = 0.45) in the participants’ pain score following a single session of the study interventions (VR or telehealth) or the control. However, a ‘medium-to-large’ group x time interaction effect (η2 = 0.10) was detected, indicating a more favorable pain score change following a single session of telehealth- (pre-post ∆: +10 ± 12 mm) and VR-delivered exercise (pre-post ∆: +9 ± 24 mm) compared to the control group (pre-post ∆: +16 ± 12 mm). Conclusions: Our study suggests that a single bout of a ‘self-managed’ VR-delivered exercise may be as efficacious as a single session of ‘supervised’ telehealth-delivered exercise in providing immediate relief from pelvic pain associated with endometriosis.
Research on the Characteristics of Safety Culture and Obstacle Factors among Residents under the Influence of COVID-19 in China
Authors: Wang Qifei , Zhao Yihan , Wang Jian , Liu Haolin , Zhang Hui
Published: January 2023

This study established a comprehensive evaluation indicator model for the safety culture among residents during COVID-19 and an obstacle degree model for the identification of the major factors affecting the residents’ safety culture. The results show that the overall level of the safety culture among residents was 0.6059. Safety education, channels for learning knowledge regarding safety, and implementation of safety management systems are currently the major obstacles affecting safety culture among residents, but there is still space for improvement in the future. Furthermore, the level of safety culture was strongly related to the distance from the infected, because this changes the risk of viral infection. There are also differences in obstacle factors in different regions. Therefore, it is necessary to implement measures targeting the improvement of safety culture in accordance with the risk of viral infection. Strategies for strengthening the safety culture are also given in this study for consideration in strategic decision making with the aim of promoting the improvement of safety culture among residents, which may help to reduce the risk of infection with COVID-19 for residents.
Acupuncture Is Effective at Reducing the Risk of Stroke in Patients with Migraines: A Real-World, Large-Scale Cohort Study with 19-Years of Follow-Up
Authors: Liao Chung-Chih , Chien Chi-Hsien , Shih Ying-Hsiu , Tsai Fuu-Jen , Li Jung-Miao
Published: January 2023

Migraines are common headache disorders and risk factors for subsequent strokes. Acupuncture has been widely used in the treatment of migraines; however, few studies have examined whether its use reduces the risk of strokes in migraineurs. This study explored the long-term effects of acupuncture treatment on stroke risk in migraineurs using national real-world data. We collected new migraine patients from the Taiwan National Health Insurance Research Database (NHIRD) from 1 January 2000 to 31 December 2017. Using 1:1 propensity-score matching, we assigned patients to either an acupuncture or non-acupuncture cohort and followed up until the end of 2018. The incidence of stroke in the two cohorts was compared using the Cox proportional hazards regression analysis. Each cohort was composed of 1354 newly diagnosed migraineurs with similar baseline characteristics. Compared with the non-acupuncture cohort, the acupuncture cohort had a significantly reduced risk of stroke (adjusted hazard ratio, 0.4; 95% confidence interval, 0.35–0.46). The Kaplan–Meier model showed a significantly lower cumulative incidence of stroke in migraine patients who received acupuncture during the 19-year follow-up (log-rank test, p < 0.001). Acupuncture confers protective benefits on migraineurs by reducing the risk of stroke. Our results provide new insights for clinicians and public health experts.
Free Choice Navigation in the Real World: Giving Back Freedom to Wayfinders
Authors: Mazurkiewicz Bartosz , Kattenbeck Markus , Giannopoulos Ioannis
Published: January 2023

In recent years, there has been collected evidence suggesting that increased usage of navigation assistance systems has a harmful effect on spatial cognition, including spatial knowledge acquisition. Previously, we proposed a potential remedy called Free Choice Navigation (simulation study). This novel navigation approach aims to provide the user with more freedom while navigating, and simultaneously give fewer navigation instructions. This approach also aims at increasing engagement with the environment and fostering spatial knowledge acquisition. We conducted a human-subject study with 48 participants comparing Free Choice Navigation with the widespread Turn-by-Turn approach on the outskirts of Vienna, Austria. The study showed the viability of our navigation system in real urban environments, providing fewer navigation instructions compared to the Turn-by-Turn approach (relative to the number of traversed junctions). Fewer instructions and forced engagement with the environment, however, did not result in differences concerning spatial knowledge acquisition, but interestingly, Free Choice Navigation users (without a map) could extract spatial configuration information similarly well as Turn-by-Turn users having a map. Moreover, we provide evidence that people are interested in learning more about their environments and are willing to walk longer routes to achieve it.
Molecular Dynamics Simulation as a Tool to Identify Mutual Synergistic Folding Proteins
Authors: Magyar Csaba , Németh Bálint Zoltán , Cserző Miklós , Simon István
Published: January 2023

Mutual synergistic folding (MSF) proteins belong to a recently emerged subclass of disordered proteins, which are disordered in their monomeric forms but become ordered in their oligomeric forms. They can be identified by experimental methods following their unfolding, which happens in a single-step cooperative process, without the presence of stable monomeric intermediates. Only a limited number of experimentally validated MSF proteins are accessible. The amino acid composition of MSF proteins shows high similarity to globular ordered proteins, rather than to disordered ones. However, they have some special structural features, which makes it possible to distinguish them from globular proteins. Even in the possession of their oligomeric three-dimensional structure, classification can only be performed based on unfolding experiments, which are frequently absent. In this work, we demonstrate a simple protocol using molecular dynamics simulations, which is able to indicate that a protein structure belongs to the MSF subclass. The presumption of the known atomic resolution quaternary structure is an obvious limitation of the method, and because of its high computational time requirements, it is not suitable for screening large databases; still, it is a valuable in silico tool for identification of MSF proteins.
Immunologic Signatures across Molecular Subtypes and Potential Biomarkers for Sub-Stratification in Endometrial Cancer
Authors: Jiang Fang , Jiang Shiyang , Cao Dongyan , Mao Mingyi , Xiang Yang
Published: January 2023

Current molecular classification approaches for endometrial cancer (EC) often employ multiple testing platforms. Some subtypes still lack univocal prognostic significance, highlighting the need for risk sub-stratification. The tumor immune microenvironment (TIME) is associated with tumor progression and prognosis. We sought to investigate the feasibility of classifying EC via DNA sequencing and interrogate immunologic signatures and prognostic markers across and within subtypes, respectively. Formalin-fixed paraffin-embedding (FFPE) samples from 50 EC patients underwent targeted DNA and RNA sequencing, and multiplex immunofluorescence assay for TIME. DNA sequencing classified 10%, 20%, 52%, and 18% of patients into the subtype of POLE-mutant, microsatellite instability-high (MSI-H), TP53-wt, and TP53-mutant. POLE-mutant tumors expressed the highest T-effector and IFN-γ signature and the lowest innate anti-PD-1 resistance signature among subtypes. TP53-wt revealed a converse enrichment trend for these immunologic signatures. Survival analyses using the Cancer Genome Atlas Uterine Corpus Endometrial Carcinoma (TCGA-UCEC) dataset identified associations of CCR5 (hazard ratio (HR) = 0.71, p = 0.035), TNFRSF14 (HR = 0.58, p = 0.028), and IL-10 (HR = 2.5, p = 0.012) with overall survival within MSI-H, TP53-mutant, and TP53-wt subtype, respectively. A TIME comparison between the sub-stratified subgroups of our cohort revealed upregulated tumor infiltration of immune cells in the low-risk subgroups. Our study demonstrates that targeted DNA sequencing is an effective one-stop strategy to classify EC. Immunomodulatory genes may serve as prognostic markers within subtypes.
Molecular Mechanisms in Genetic Aortopathy–Signaling Pathways and Potential Interventions
Authors: Dong Charlotte Xue , Malecki Cassandra , Robertson Elizabeth , Hambly Brett , Jeremy Richmond
Published: January 2023

Thoracic aortic disease affects people of all ages and the majority of those aged <60 years have an underlying genetic cause. There is presently no effective medical therapy for thoracic aneurysm and surgery remains the principal intervention. Unlike abdominal aortic aneurysm, for which the inflammatory/atherosclerotic pathogenesis is well established, the mechanism of thoracic aneurysm is less understood. This paper examines the key cell signaling systems responsible for the growth and development of the aorta, homeostasis of endothelial and vascular smooth muscle cells and interactions between pathways. The evidence supporting a role for individual signaling pathways in pathogenesis of thoracic aortic aneurysm is examined and potential novel therapeutic approaches are reviewed. Several key signaling pathways, notably TGF-β, WNT, NOTCH, PI3K/AKT and ANGII contribute to growth, proliferation, cell phenotype and survival for both vascular smooth muscle and endothelial cells. There is crosstalk between pathways, and between vascular smooth muscle and endothelial cells, with both synergistic and antagonistic interactions. A common feature of the activation of each is response to injury or abnormal cell stress. Considerable experimental evidence supports a contribution of each of these pathways to aneurysm formation. Although human information is less, there is sufficient data to implicate each pathway in the pathogenesis of human thoracic aneurysm. As some pathways i.e., WNT and NOTCH, play key roles in tissue growth and organogenesis in early life, it is possible that dysregulation of these pathways results in an abnormal aortic architecture even in infancy, thereby setting the stage for aneurysm development in later life. Given the fine tuning of these signaling systems, functional polymorphisms in key signaling elements may set up a future risk of thoracic aneurysm. Multiple novel therapeutic agents have been developed, targeting cell signaling pathways, predominantly in cancer medicine. Future investigations addressing cell specific targeting, reduced toxicity and also less intense treatment effects may hold promise for effective new medical treatments of thoracic aortic aneurysm.
Inhibition of Voltage-Gated Na+ Currents Exerted by KB-R7943 (2-[2-[4-(4-nitrobenzyloxy)phenyl]ethyl]isothiourea), an Inhibitor of Na+-Ca2+ Exchanging Process
Authors: Wu Sheng-Nan , Yu Meng-Cheng
Published: January 2023

KB-R7943, an isothiourea derivative, has been recognized as an inhibitor in the reverse mode of the Na+-Ca2+ exchanging process. This compound was demonstrated to prevent intracellular Na+-dependent Ca2+ uptake in intact cells; however, it is much less effective at preventing extracellular Na+-dependent Ca2+ efflux. Therefore, whether or how this compound may produce any perturbations on other types of ionic currents, particularly on voltage-gated Na+ current (INa), needs to be further studied. In this study, the whole-cell current recordings demonstrated that upon abrupt depolarization in pituitary GH3 cells, the exposure to KB-R7943 concentration-dependently depressed the transient (INa(T)) or late component (INa(L)) of INa with an IC50 value of 11 or 0.9 μM, respectively. Likewise, the dissociation constant for the KB-R7943-mediated block of INa on the basis of a minimum reaction scheme was estimated to be 0.97 μM. The presence of benzamil or amiloride could suppress the INa(L) magnitude. The instantaneous window Na+ current (INa(W)) activated by abrupt ascending ramp voltage (Vramp) was suppressed by adding KB-R7943; however, subsequent addition of deltamethrin or tefluthrin (Tef) effectively reversed KB-R7943-inhibted INa(W). With prolonged duration of depolarizing pulses, the INa(L) amplitude became exponentially decreased; moreover, KB-R7943 diminished INa(L) magnitude. The resurgent Na+ current (INa(R)) evoked by a repolarizing Vramp was also suppressed by adding this compound; moreover, subsequent addition of ranolazine or Tef further diminished or reversed, respectively, its reduction in INa(R) magnitude. The persistent Na+ current (INa(P)) activated by sinusoidal voltage waveform became enhanced by Tef; however, subsequent application of KB-R7943 counteracted Tef-stimulated INa(P). The docking prediction reflected that there seem to be molecular interactions of this molecule with the hNaV1.2 or hNaV1.7 channels. Collectively, this study highlights evidence showing that KB-R7943 has the propensity to perturb the magnitude and gating kinetics of INa (e.g., INa(T), INa(L), INa(W), INa(R), and INa(P)) and that the NaV channels appear to be important targets for the in vivo actions of KB-R7943 or other relevant compounds.
Neurotrophins in the Neuropathophysiology, Course, and Complications of Obstructive Sleep Apnea—A Narrative Review
Authors: Gabryelska Agata , Turkiewicz Szymon , Ditmer Marta , Sochal Marcin
Published: January 2023

Obstructive sleep apnea (OSA) is a disorder characterized by chronic intermittent hypoxia and sleep fragmentation due to recurring airway collapse during sleep. It is highly prevalent in modern societies, and due to its pleiotropic influence on the organism and numerous sequelae, it burdens patients and physicians. Neurotrophins (NTs), proteins that modulate the functioning and development of the central nervous system, such as brain-derived neurotrophic factor (BDNF), have been associated with OSA, primarily due to their probable involvement in offsetting the decline in cognitive functions which accompanies OSA. However, NTs influence multiple aspects of biological functioning, such as immunity. Thus, extensive evaluation of their role in OSA might enlighten the mechanism behind some of its elusive features, such as the increased risk of developing an immune-mediated disease or the association of OSA with cardiovascular diseases. In this review, we examine the interactions between NTs and OSA and discuss their contribution to OSA pathophysiology, complications, as well as comorbidities.
Overview of the Role of Vanillin in Neurodegenerative Diseases and Neuropathophysiological Conditions
Authors: Iannuzzi Clara , Liccardo Maria , Sirangelo Ivana
Published: January 2023

Nowadays, bioactive natural products play key roles in drug development due to their safety profile and strong antioxidant power. Vanillin is a natural phenolic compound found in several vanilla beans and widely used for food, cosmetic, and pharmaceutical products. Besides its industrial applications, vanillin possesses several beneficial effects for human health, such as antioxidant activity in addition to anti-inflammatory, anti-mutagenic, anti-metastatic, and anti-depressant properties. Moreover, vanillin exhibits neuroprotective effects on multiple neurological disorders and neuropathophysiological conditions. This study reviews the mechanisms of action by which vanillin prevents neuroinflammation and neurodegeneration in vitro and in vivo systems, in order to provide the latest views on the beneficial properties of this molecule in chronic neurodegenerative diseases and neuropathophysiological conditions.
Association of the Telomerase Reverse Transcriptase rs10069690 Polymorphism with the Risk, Age at Onset and Prognosis of Triple Negative Breast Cancer
Authors: Zins Karin , Peka Elisabeth , Miedl Heidi , Ecker Stefanie , Abraham Dietmar , Schreiber Martin
Published: January 2023

Telomerase reverse transcriptase (TERT) plays a key role in the maintenance of telomere DNA length. The rs10069690 single nucleotide variant, located in intron 4 of TERT, was found to be associated with telomere length and the risk of estrogen receptor-negative but not–positive breast cancer. This study aimed at analysis of the association of rs10069690 genotype and TERT expression with the risk, age at onset, prognosis, and clinically and molecularly relevant subtypes of breast cancer. Accordingly, rs10069690 was genotyped in a hospital-based case-control study of 403 female breast cancer patients and 246 female controls of a Central European (Austrian) study population, and the mRNA levels of TERT were quantified in 106 primary breast tumors using qRT-PCR. We found that in triple-negative breast cancer patients, the minor rs10069690 TT genotype tended to be associated with an increased breast cancer risk (OR, 1.87; 95% CI, 0.75–4.71; p = 0.155) and was significantly associated with 11.7 years younger age at breast cancer onset (p = 0.0002), whereas the CC genotype was associated with a poor brain metastasis-free survival (p = 0.009). Overall, our data show that the rs10069690 CC genotype and a high TERT expression tended to be associated with each other and with a poor prognosis. Our findings indicate a key role of rs10069690 in triple-negative breast cancer.
Identification of Pre-Renal and Intrinsic Acute Kidney Injury by Anamnestic and Biochemical Criteria: Distinct Association with Urinary Injury Biomarkers
Authors: Sancho-Martínez Sandra M. , Casanova Alfredo G. , Düwel Annette G. , Rivero-García Karen , García-Garrido Tamara , Morales Ana I. , Martínez-Salgado Carlos , López-Hernández Francisco J. , Fraile Pilar
Published: January 2023

Acute kidney injury (AKI) is a syndrome of sudden renal excretory dysfunction with severe health consequences. AKI etiology influences prognosis, with pre-renal showing a more favorable evolution than intrinsic AKI. Because the international diagnostic criteria (i.e., based on plasma creatinine) provide no etiological distinction, anamnestic and additional biochemical criteria complement AKI diagnosis. Traditional, etiology-defining biochemical parameters, including the fractional excretion of sodium, the urinary-to-plasma creatinine ratio and the renal failure index are individually limited by confounding factors such as diuretics. To minimize distortion, we generated a composite biochemical criterion based on the congruency of at least two of the three biochemical ratios. Patients showing at least two ratios indicative of intrinsic AKI were classified within this category, and those with at least two pre-renal ratios were considered as pre-renal AKI patients. In this study, we demonstrate that the identification of intrinsic AKI by a collection of urinary injury biomarkers reflective of tubular damage, including NGAL and KIM-1, more closely and robustly coincide with the biochemical than with the anamnestic classification. Because there is no gold standard method for the etiological classification of AKI, the mutual reinforcement provided by the biochemical criterion and urinary biomarkers supports an etiological diagnosis based on objective diagnostic parameters.
Lipopolysaccharide Tolerance Enhances Murine Norovirus Reactivation: An Impact of Macrophages Mainly Evaluated by Proteomic Analysis
Authors: Makjaroen Jiradej , Phuengmaung Pornpimol , Saisorn Wilasinee , Udomkarnjananun Suwasin , Pisitkun Trairak , Leelahavanichkul Asada
Published: January 2023

Because of endotoxemia during sepsis (a severe life-threatening infection), lipopolysaccharide (LPS) tolerance (the reduced responses to the repeated LPS stimulation) might be one of the causes of sepsis-induced immune exhaustion (the increased susceptibility to secondary infection and/or viral reactivation). In LPS tolerance macrophage (twice-stimulated LPS, LPS/LPS) compared with a single LPS stimulation (N/LPS), there was (i) reduced energy of the cell in both glycolysis and mitochondrial activities (extracellular flux analysis), (ii) decreased abundance of the following proteins (proteomic analysis): (a) complex I and II of the mitochondrial electron transport chain, (b) most of the glycolysis enzymes, (c) anti-viral responses with Myxovirus resistance protein 1 (Mx1) and Ubiquitin-like protein ISG15 (Isg15), (d) antigen presentation pathways, and (iii) the down-regulated anti-viral genes, such as Mx1 and Isg15 (polymerase chain reaction). To test the correlation between LPS tolerance and viral reactivation, asymptomatic mice with and without murine norovirus (MNV) infection as determined in feces were tested. In MNV-positive mice, MNV abundance in the cecum, but not in feces, of LPS/LPS mice was higher than that in N/LPS and control groups, while MNV abundance of N/LPS and control were similar. Additionally, the down-regulated Mx1 and Isg15 were also demonstrated in the cecum, liver, and spleen in LPS/LPS-activated mice, regardless of MNV infection, while N/LPS more prominently upregulated these genes in the cecum of MNV-positive mice compared with the MNV-negative group. In conclusion, defects in anti-viral responses after LPS tolerance, perhaps through the reduced energy status of macrophages, might partly be responsible for the viral reactivation. More studies on patients are of interest.
Detectable Lipidomes and Metabolomes by Different Plasma Exosome Isolation Methods in Healthy Controls and Patients with Advanced Prostate and Lung Cancer
Authors: Soupir Alex C. , Tian Yijun , Stewart Paul A. , Nunez-Lopez Yury O. , Manley Brandon J. , Pellini Bruna , Bloomer Amanda M. , Zhang Jingsong , Mo Qianxing , Marchion Douglas C. , Liu Min , Koomen John M. , Siegel Erin M. , Wang Liang
Published: January 2023

Circulating exosomes in the blood are promising tools for biomarker discovery in cancer. Due to their heterogeneity, different isolation methods may enrich distinct exosome cargos generating different omic profiles. In this study, we evaluated the effects of plasma exosome isolation methods on detectable multi-omic profiles in patients with non-small cell lung cancer (NSCLC), castration-resistant prostate cancer (CRPC), and healthy controls, and developed an algorithm to quantify exosome enrichment. Plasma exosomes were isolated from CRPC (n = 10), NSCLC (n = 14), and healthy controls (n = 10) using three different methods: size exclusion chromatography (SEC), lectin binding, and T-cell immunoglobulin domain and mucin domain-containing protein 4 (TIM4) binding. Molecular profiles were determined by mass spectrometry of extracted exosome fractions. Enrichment analysis of uniquely detected molecules was performed for each method with MetaboAnalyst. The exosome enrichment index (EEI) scores methods based on top differential molecules between patient groups. The lipidomic analysis detected 949 lipids using exosomes from SEC, followed by 246 from lectin binding and 226 from TIM4 binding. The detectable metabolites showed SEC identifying 191 while lectin binding and TIM4 binding identified 100 and 107, respectively. When comparing uniquely detected molecules, different methods showed preferential enrichment of different sets of molecules with SEC enriching the greatest diversity. Compared to controls, SEC identified 28 lipids showing significant difference in NSCLC, while only 1 metabolite in NSCLC and 5 metabolites in CRPC were considered statistically significant (FDR < 0.1). Neither lectin-binding- nor TIM4-binding-derived exosome lipids or metabolites demonstrated significant differences between patient groups. We observed the highest EEI from SEC in lipids (NSCLC: 871.33) which was also noted in metabolites. These results support that the size exclusion method of exosome extraction implemented by SBI captures more heterogeneous exosome populations. In contrast, lectin-binding and TIM4-binding methods bind surface glycans or phosphatidylserine moieties of the exosomes. Overall, these findings suggest that specific isolation methods select subpopulations which may significantly impact cancer biomarker discovery.
Mitochondrial Metabolism in X-Irradiated Cells Undergoing Irreversible Cell-Cycle Arrest
Authors: Hirose Eri , Noguchi Miho , Ihara Tomokazu , Yokoya Akinari
Published: January 2023

Irreversible cell-cycle-arrested cells not undergoing cell divisions have been thought to be metabolically less active because of the unnecessary consumption of energy for cell division. On the other hand, they might be actively involved in the tissue microenvironment through an inflammatory response. In this study, we examined the mitochondria-dependent metabolism in human cells irreversibly arrested in response to ionizing radiation to confirm this possibility. Human primary WI-38 fibroblast cells and the BJ-5ta fibroblast-like cell line were exposed to 20 Gy X-rays and cultured for up to 9 days after irradiation. The mitochondrial morphology and membrane potential were evaluated in the cells using the mitochondrial-specific fluorescent reagents MitoTracker Green (MTG) and 5,5′,6,6′-tetraethyl-benzimidazolylcarbocyanine iodide (JC-1), respectively. The ratio of the mean MTG-stained total mitochondrial area per unit cell area decreased for up to 9 days after X-irradiation. The fraction of the high mitochondrial membrane potential area visualized by JC-1 staining reached its minimum 2 days after irradiation and then increased (particularly, WI-38 cells increased 1.8-fold the value of the control). Their chronological changes indicate that the mitochondrial volume in the irreversible cell-cycle-arrested cells showed significant increase concurrently with cellular volume expansion, indicating that the mitochondria-dependent energy metabolism was still active. These results indicate that the energy metabolism in X-ray-induced senescent-like cells is active compared to nonirradiated normal cells, even though they do not undergo cell divisions.
Favorable Prognosis in Patients with Recovered Pulmonary Hypertension after TAVI: An Analysis of the LAPLACE-TAVI Registry
Authors: Koike Takuma , Iwata Hiroshi , Chikata Yuichi , Doi Shinichiro , Naito Ryo , Yasuda Hidetoshi , Funamizu Takehiro , Endo Hirohisa , Miyazaki Sakiko , Okazaki Shinya , Higuchi Ryosuke , Takamisawa Itaru , Sato Kei , Tamura Harutoshi , Yokoyama Hiroaki , Tobaru Tetsuya , Takanashi Shuichiro , Tabata Minoru , Minamino Tohru
Published: January 2023

Pulmonary hypertension (PH) is a common complication of aortic stenosis (AS). Despite the established association between PH and poor outcomes in patients with AS, the prognostic implication of a change in PH after transcatheter aortic valve implantation (TAVI) has been rarely evaluated. This study analyzed a prospective multi-center TAVI registry database involving six Japanese centers and used the transtricuspid pressure gradient (TRPG) obtained by echocardiography to estimate pulmonary artery systolic pressure. The participants (n = 2056) were first divided into two groups by TRPG before TAVI, a PH (−) group (TRPG < 30 mmHg) (n = 1407, 61.9%) and a PH (+) group (TRPG ≥ 30 mmHg) (n = 649, 28.6%). Next, by TRPG after (4.1 ± 5.3 days) TAVI, the PH (+) group was further subdivided into two groups, Recovered PH (TRPG < 30 mmHg, n = 253) and Persistent PH (TRPG after TAVI ≥ 30 mmHg, n = 396). The median follow-up duration was 1.8 years. The primary and secondary endpoints were the composite and each of cardiovascular (CV) death and heart failure hospitalization, respectively. Unadjusted Kaplan-Meier estimates with log-rank comparisons showed significantly higher cumulative incidences of primary and secondary endpoints in the Persistent PH group compared to other groups. Moreover, adjusted multivariate Cox-proportional hazard analyses showed that a decreased (−10 mmHg) TRPG after TAVI was linearly associated with a reduced risk of the primary endpoint (hazard ratio (HR): 0.76, 95% confidence interval (CI): 0.64–0.90, p = 0.0020). The findings in the present study indicate that the recovery of PH may partly contributes to the prognostic benefit of TAVI procedure in patients with AS and elevated pulmonary artery systolic pressure.
Surgical Techniques of Gastrojejunostomy in Robotic Pancreatoduodenectomy: Robot-Sewn versus Stapled Gastrojejunostomy Anastomosis
Authors: Takagi Kosei , Umeda Yuzo , Yoshida Ryuichi , Fuji Tomokazu , Yasui Kazuya , Kimura Jiro , Hata Nanako , Yagi Takahito , Fujiwara Toshiyoshi
Published: January 2023

Background: Delayed gastric emptying (DGE) is a major complication of pancreatoduodenectomy (PD). Several efforts have been made to decrease the incidence of DGE. However, the optimal anastomotic method for gastro/duodenojejunostomy (GJ) remains debatable. Moreover, few studies have reported the impact of GJ surgical techniques on outcomes following robotic pancreatoduodenectomy (RPD). This study aimed to investigate the surgical outcomes of robot-sewn and stapled GJ anastomoses in RPD. Methods: Forty patients who underwent RPD at the Okayama University Hospital between September 2020 and October 2022 were included. The outcomes between robot-sewn and stapled anastomoses were compared. Results: The mean [standard deviation (SD)] operative and GJ time were 428 (63.5) and 34.0 (15.0) minutes, respectively. Postoperative outcomes included an overall incidence of DGE of 15.0%, and the mean postoperative hospital stays were 11.6 (5.3) days in length. The stapled group (n = 21) had significantly shorter GJ time than the robot-sewn group (n = 19) (22.7 min versus 46.5 min, p < 0.001). Moreover, stapled GJ cases were significantly associated with a lower incidence of DGE (0% versus 21%, p = 0.01). Although not significant, the stapled group tended to have shorter postoperative hospital stays (9.9 days versus 13.5 days, p = 0.08). Conclusions: Our findings suggest that stapled GJ anastomosis might decrease anastomotic GJ time and incidence of DGE after RPD. Surgeons should select a suitable method for GJ anastomosis based on their experiences with RPD.
Association between Early Immune-Related Adverse Events and Survival in Patients Treated with PD-1/PD-L1 Inhibitors
Authors: Zhang You-Cheng , Zhu Tian-Chen , Nie Run-Cong , Lu Liang-He , Xiang Zhi-Cheng , Xie Dan , Luo Rong-Zhen , Cai Mu-Yan
Published: January 2023

Background: Immune-related adverse events (irAEs) are side effects that reflect the activation of patients’ immune systems after treatment with immune checkpoint inhibitors (ICIs). However, there is no meta-analysis on the effect of early irAEs on patient survival. Thus, we assessed the association between early irAEs and the survival of patients treated with ICIs. Methods: PubMed, Embase, and Web of Science were searched from May 2010 to May 2020 for all the retrospective and prospective comparative studies to evaluate the hazard ratios (HRs) for death. A random-effects model was used to calculate the pooled HR for death, and heterogeneity was assessed using I² statistics. The main outcomes were overall survival (OS) and progression-free survival (PFS). Results: A total of 11 reports with 2077 patients were included. A significant association was observed between early irAEs and a favorable clinical outcome. Patients with early irAEs had prolonged OS (HR: 0.62, 95% confidence interval (CI): 0.53–0.74, p < 0.001) and PFS (HR: 0.53, 95% CI: 0.41–0.66, p < 0.001) compared to those without; these results were confirmed using a sensitivity analysis. The irAE types, malignancy types, and sample size were correlated with patients’ clinical outcomes. Conclusions: Early irAEs, especially cutaneous irAEs, correlated with a better clinical outcome in patients treated with ICIs.
Long-Term Results after Autologous Breast Reconstruction with DIEP versus PAP Flaps Based on Quality of Life and Aesthetic Outcome Analysis
Authors: Augustin Angela , Morandi Evi M. , Winkelmann Selina , Schoberleitner Ines , Egle Daniel , Ritter Magdalena , Bauer Thomas , Wachter Tanja , Wolfram Dolores
Published: January 2023

(1) Background: This work aimed to conduct a comparative study, providing long-term data about patient-reported outcome measures and donor site scar assessments, as well as an aesthetic evaluation of the reconstructed breasts in patients with DIEP versus PAP flap breast reconstruction. (2) Methods: This prospective, single-center, matched cohort study included a total of 36 patients after DIEP and PAP flap breast reconstruction. The evaluation was carried out using the Breast-Q and POSAS questionnaire, as well as the Breast Aesthetic Scale for cosmetic analysis, by four plastic surgeons. (3) Results: The postoperative Breast-Q evaluation revealed no significant differences between both patient groups for the categories of the physical well-being of the donor site, the physical well-being of the breast, and satisfaction with the breast. A scar evaluation of the donor site region showed equivalent results for the thigh and abdomen regions, concerning the overall opinion of the patients and the observers. There was no significant difference between both methods of reconstruction for all aspects of breast aesthetics. (4) Conclusions: Similar results for donor site morbidity, scar quality, and the aesthetic outcome of the breasts in both the DIEP and PAP patient groups have been demonstrated. Hence, in those cases suitable for both types of reconstruction, the decision can be based on factors such as patients’ lifestyles, leisure activities, and preferences.
Clinical-Radiological Mismatch in Multiple Sclerosis Patients during Acute Relapse: Discrepancy between Clinical Symptoms and Active, Topographically Fitting MRI Lesions
Authors: Dünschede Jutta , Ruschil Christoph , Bender Benjamin , Mengel Annerose , Lindig Tobias , Ziemann Ulf , Kowarik Markus C.
Published: January 2023

Background: Relapses in multiple sclerosis (MS) patients are usually defined as subacute clinical symptoms that last for at least 24 h. To validate a clinical relapse on magnetic resonance imaging (MRI), an anatomically fitting lesion with gadolinium enhancement in the central nervous system (CNS) would be mandatory. The aim of this study was to validate clinical relapses in regard to the concomitant detection of active, anatomically fitting MRI lesions. Methods: We performed a retrospective analysis of 199 MS patients with acute relapse who had received an MRI scan before the initiation of methylprednisolone (MPS) therapy. Clinical data and MRIs were systematically reanalyzed by correlating clinical symptoms with their anatomical representation in the CNS. Patients were then categorized into subgroups with a clinical-radiological match (group 1) or clinical-radiological mismatch (group 2) between symptoms and active, topographically fitting lesions and further analyzed in regard to clinical characteristics. Results: In 43% of our patients, we observed a clinical-radiological mismatch (group 2). Further analysis of patient characteristics showed that these patients were significantly older at the time of relapse. MS patients in group 2 also showed a significantly longer disease duration and significantly more previous relapses when compared to group 1. Comparing symptom clusters, the appearance of motor dysfunction during the current relapse was significantly more frequent in group 2 than in group 1. The overall dose of MPS treatment was significantly lower in group 2 than in group 1 with a similar treatment response in both groups. Conclusions: The substantial clinical-radiological mismatch during acute relapse in our study could be explained by several factors, including a psychosomatic component or disturbance of network connectivity. Alternatively, secondary progression or a diffuse neuro-inflammatory process might cause clinical symptoms, especially in older patients with a longer disease duration. As a consequence, treatment of clinical relapses and the definition of breakthrough disease should be reconsidered in regard to combined clinical and MRI criteria and/or additional biomarkers. Further studies are necessary to address the contribution of diffuse neuro-inflammation to the clinical presentation of symptoms.