bluuebunny/crossref_metadata_2025 huggingface dataset:
Escalated‐dose radiotherapy for unresected locally advanced pancreatic cancer: Patterns of care and survival in the United States
Authors: Shi Christopher , De Brian , Tran Cao Hop S. , Liu Suyu , Florez Marcus A. , Kouzy Ramez , Grippin Adam J. , Katz Matthew H. G. , Tzeng Ching‐Wei D. , Ikoma Naruhiko , Kim Michael P. , Lee Sunyoung , Willis Jason , Noticewala Sonal S. , Minsky Bruce D. , Smith Grace L. , Holliday Emma B. , Taniguchi Cullen M. , Koong Albert C. , Das Prajnan , Ludmir Ethan B. , Koay Eugene J.
Published: June 2024

Introduction

With locally advanced pancreatic cancer (LAPC), uncontrolled local tumor growth frequently leads to mortality. Advancements in radiotherapy (RT) techniques have enabled conformal delivery of escalated‐dose RT (EDR), which may have potential local control and overall survival (OS) benefits based on retrospective and early prospective studies. With evidence for EDR emerging, we characterized the adoption of EDR across the United States and its associated outcomes.

Methods

We searched the National Cancer Database for nonsurgically managed LAPC patients diagnosed between 2004 and 2019. Pancreas‐directed RT with biologically effective doses (BED10) ≥39 and ≤70 Gy was labeled conventional‐dose RT (CDR), and BED10 >70 and ≤132 Gy was labeled EDR. We identified associations of EDR and OS using logistic and Cox regressions, respectively.

Results

Among the definitive therapy subset (n = 54,115) of the entire study cohort (n = 91,493), the most common treatments were chemotherapy alone (69%), chemotherapy and radiation (29%), and RT alone (2%). For the radiation therapy subset (n = 16,978), use of pancreas‐directed RT remained between 13% and 17% over the study period (ptrend > 0.999). Using multivariable logistic regression, treatment at an academic/research facility (adjusted odds ratio [aOR] 1.46, p < 0.001) and treatment between 2016 and 2019 (aOR 2.54, p < 0.001) were associated with greater receipt of EDR, whereas use of chemotherapy (aOR 0.60, p < 0.001) was associated with less receipt. Median OS estimates for EDR and CDR were 14.5 months and 13.0 months (p < 0.0001), respectively. For radiation therapy subset patients with available survival data (n = 13,579), multivariable Cox regression correlated EDR (adjusted hazard ratio 0.85, 95% confidence interval 0.80–0.91; p < 0.001) with longer OS versus CDR.

Discussion and Conclusions

Utilization of EDR has increased since 2016, but overall utilization of RT for LAPC has remained at less than one in five patients for almost two decades. These real‐world results additionally provide an estimate of effect size of EDR for future prospective trials.
The relationship between lifelong learning and professional development activities in a Portuguese sample
Authors: Coelho Rodrigo Ponces , Pires António Pazo , Neimeyer Greg , Vaz Alexandre , Neto David , Sousa Daniel
Published: June 2024

Objective

A dispositional commitment to lifelong learning (LL) is an important aspect of continuing professional development for practicing psychotherapists. Equally important is the dedication of psychotherapists to engaging in professional development activities to foster growth and maintain competence. However, there are limited available data on the range of practices and factors influencing psychotherapists' commitment to these activities. The primary objective of this study was to examine whether a higher commitment to LL would significantly predict greater participation in a range of different professional development activities.

Method

A sample of 166 Portuguese psychotherapists completed an online survey on professional development. The survey included two key measures: the Jefferson Scale of Psychotherapist Lifelong Learning and a list of professional development activities based on the Retrospective Analysis of Psychotherapists' Involvement in Deliberate Practice. Exploratory factorial analysis was conducted to determine an alternative and suitable grouping for the list of activities, and structural equation modelling was utilised to assess the relationship between the commitment to LL and engagement in these activities.

Findings

Results provided valuable insight into the diverse levels of participation among therapists across five categories of activities: (1) supervision and case discussion; (2) recording‐related activities; (3) reading and focussed learning; (4) reflecting; and (5) workshops supporting ongoing professional competence. Findings indicated that a stronger commitment to LL significantly predicted increased dedication to reading, focussed learning activities and attendance to training workshops. The implications and limitations of the study are discussed concerning their practical application for professional psychologists, counsellors and psychotherapists.
Antioxidant Mechanism, Spectroscopic and Pharmacological Properties of Four Flavonoids: DFT, Docking and Molecular Dynamics
Authors: Lai Wu‐Ji , Lu Jia‐Hao , Chen Wen‐hao , Jiang Li‐he , Shen Li‐Qun
Published: June 2024

Myricetin (1), Quercetin (2), Kaempferol (3) and Kaempferide (4) were flavonoids with phenolic hydroxyl groups. The antioxidant and pharmacological mechanisms of them were investigated in detail. The lowest hydroxyl dissociation enthalpies of 1, 2, 3 and 4 were calculated by DFT, respectively. The hydroxyl dissociation enthalpies of the four flavonoids at the O2 site are the highest. By analyzing the intramolecular hydrogen bonds and HOMO‐LUMO orbitals of the four flavonoids, the reasons for their divergence of hydroxyl dissociation enthalpies and antioxidant mechanisms were further investigated. The UV–vis and IR spectra of four flavonoids were compared. The interactions about electrostatic attraction, p–π conjugation and hydrogen bond combined the flavonoid with the target protein closely. The root mean square deviation of peroxisome proliferator‐activated receptor γ combined with 1, 2 and 3 increased, while that of PPARγ combined with 4 decreased.
Protective Effect of a Isothiazolinone Derivative on Acute Lung Injury by Regulating PI3K‐AKT Signaling Pathway
Authors: Li Hua , Guo Jie , Zhang Gaiyue , Zhou Jing , Wang Qiang
Published: June 2024

Acute lung injury (ALI) is a prevalent organ injury in sepsis, characterized by an inflammatory reactive disorder. Both the incidence and mortality rates of ALI have been steadily increasing. Isothiazolinone derivatives have displayed anti‐inflammatory activity and have shown effectiveness in treating pneumonia. The objective of the study is to assess the effects and mechanisms of the isothiazolinone derivative 4‐benzoyl‐2‐butyl‐5‐(ethylsulfinyl)isothiazol‐3(2H)‐one (C6) on sepsis‐induced ALI.The analysis of biological function and signal pathway enrichment demonstrated that C6 primarily exhibited anti‐inflammatory effects. Administration of different doses of C6 through intraperitoneal injection significantly improved the survival rate, body temperature, and body mass of mice with ALI induced by cecal ligation and puncture (CLP). Additionally, it mitigated lung tissue injury, pulmonary edema, lung permeability, inflammatory cell infiltration, apoptosis, and the expression of inflammatory cytokines. Network targeting analysis and experimental validation in mouse leukemia cells of monocyte macrophage (RAW264.7) cells and CLP‐induced ALI mice revealed that the anti‐inflammatory effect of C6 was mediated by the inhibition of the phosphatidylinositol 3 kinase ‐protein kinase B (PI3K‐AKT) signaling pathway.

The research suggest that C6 has protective effects against ALI by inhibiting the PI3K‐AKT signaling pathway. This information could be valuable in developing potential treatments for ALI.
GenusBerberis: A Comprehensive and Updated Review on Ethnobotanical Uses, Phytochemistry and Pharmacological Activities
Authors: Fatima Hashmi Samar , Saleem Hammad , Khurshid Umair , Khursheed Anjum , Tauquir Alam Md , Imran Mohd , Abida  , Nayeem Naira , Shoaib Ali Gill Muhammad
Published: June 2024

GenusBerberisis an excellent choice for research due to its history in traditional medicine, diverse pharmacological properties, and it has potential for drug discovery. This review presents information on the ethnobotany, pharmacological activities, and many phytochemicals identified fromBerberisspecies. It examines the existing literature on the genusBerberis, drawn from online databases, including PubMed, Web of Science, Science Direct, Elsevier, and Google Scholar, etc encompassing the data from 1960 to 2023. This review focuses on the structural details of reported phytochemicals ofBerberisspecies and pharmacological actions. Different extraction techniques were evaluated for extracts preparation. According to literature review, phytochemical analysis exhibited the presence of alkaloids, flavonoids, and phenolic compounds. A major bioactive alkaloid, berberine exhibits its main role in treatment of many gastric, infectious, and chronic disorders. This literature indicates thatBerberisgenus exhibits a variety of biological activities, i.e anti‐inflammatory, cytotoxic, hepatoprotective, antimicrobial, antidiabetic and antioxidant activities and utilization of these effects in the treatment and management of various diseases, like diabetes, microbial infections, inflammation, liver disorders, and cancer. However, conventional medicines, validation of traditional uses, and in‐depth phytochemical analysis are areas of research in genusBerberis.
Exploring New 5‐Nitroimidazole Derivatives As Potent Acetylcholinesterase and Butyrylcholinesterase Enzyme Inhibitors
Authors: Gursoy Sule , Satici Doruk , Kuzu Burak , Turkmenoglu Burcin , Dilek Esra , Algul Oztekin
Published: June 2024

Discovering new compounds capable of inhibiting physiologically and metabolically significant drug targets or enzymes is of paramount importance in biological chemistry. With this aim, new 5‐nitroimidazole derivatives (1–4) were designed and synthesized, and their inhibitory activities against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) were discovered using acetyl (butyryl) thiocholine and Ellman's reagents for spectrophotometric assay. The inhibitory profiles of the synthesized compounds were assessed by comparing their IC50 and Ki values. Results demonstrate significant inhibitory activity of all synthesized compounds against both AChE and BuChE compared to the reference compound, donepezil. Notably, compound 4 exhibited dual inhibition of these enzymes, showing the highest activity against Electrophorus electricus AChE (EeAChE) with a Ki value of 0.024±0.009 nM and against equine BuChE (eqBuChE) with a Ki value of 0.087±0.017 nM. Furthermore, molecular modeling was conducted to study the interaction modes of the most potent compound (4) and donepezil in the active site of their related enzymes’ crystal structures (PDB ID: 4EY7 and 4BDS, respectively). Additionally, drug‐likeness, ADME, and toxicity profiles of the compounds and metronidazole were predicted. The above results indicated that the dual inhibition of these enzymes is considered as a promising strategy for the treatment of neurological disorder especially Alzheimer's disease.
Discovery of Sesquiterpene Lactones with Cytotoxicity from the Herb of Youngia japonica
Authors: Ye Xian‐Sheng , Lin Kuan , Tao Xiao‐Qing , Shang Jin‐Ting , Gui Yu‐Ran , Zhu Shu‐Xiu , Zhou Le , Xia Yi‐Yuan , Liu Wei , Sun Bin‐Lian , Chen Hai‐Feng , Shu Xi‐Ji
Published: June 2024

In the process of searching for anti‐breast cancer agents, five sesquiterpene lactones (1‐5), including two previously undescribed ones, yjaponica B‐C (1‐2), were isolated from the herb of Youngia japonica. Their structures were elucidated by spectroscopic data analyses and Marfey's method. Cytotoxic activities of all compounds against A549, U87, and 4T1 cell lines were tested using the CCK8 assay. The result showed that compound 3 possessed the highest cytotoxic activity against 4T1 cells with an IC50 value of 10.60 μM. Furthermore, compound 3 distinctly induced apoptosis, inhibited immigration, and blocked the cell cycle of 4T1 cells. In addition, compound 3 induced the production of reactive oxygen species. Further anticancer mechanism studies showed that compound 3 significantly upregulated expression of the cleaved caspase 3 and PARP, whereas it downregulated the expression of Bcl‐2, cyclin D1, cyclin A2, CDK4, and CDK2. Taken together, our results demonstrate that compound 3 has a high potential of being used as a leading compound for the discovery of new anti‐breast cancer agent.
Clinical case of Cor triatriatum sinister, a dilemma of anticoagulation: A case report and literature review
Authors: Arzhangzade Alireza , Zamirian Mahmood , Nozhat Salma , Shafei Sasan , Narimani Javid Roozbeh , Salahi Sarvenaz , Khorshidi Soorena
Published: June 2024

Key Clinical Message

Cor triatriatum is a rare congenital heart abnormality in which a membrane separates the left atrium (LA; sinister) or the right atrium (dexter) into two compartments. It is also a long‐forgotten cause of atrial fibrillation (AF) and substantially higher rates of blood stagnation, particularly proximal to the additional septum in the LA. In this case report, we faced a CHA2DS2‐VASc score of 1 in patients with non‐valvular AF due to Cor triatriatum sinister (CTS). The decision to start anticoagulants in this particular case was controversial, so we reviewed the literature to assess and address it. We present our case and discuss the indication of anticoagulants in this unique clinical scenario, accompanied by a literature review. Facing this dilemma of starting anticoagulants in special cases of CTS and AF should be individualized and need more investigation. However, till this moment, based on similar reports, it seems to be rational to consider CTS Per se as an additional risk stratification marker beyond the CHA2DS2‐VASc score start anticoagulant until the surgical resection. Considering CTS as the sole indication of anticoagulant in patients with normal sinus rhythm is a complex matter that needs further investigation.
Nasogastric tube syndrome: A case of vocal cord paralysis due to a nasogastric tube
Authors: Shioya Nobuki , Inoue Nozomu , Kawamura Hitomi , Kawashima Naonori , Sato Tomohiro , Sadamoto Yoshihiro , Tsukamoto Yuki , Naito Yuki , Hazama Kohji , Shichinohe Yasuo
Published: June 2024

Key Clinical Message

Insertion of a nasogastric tube is one of the most common methods of administering nutrition, but can cause vocal cord paralysis.
Mechanochemical Synthesis of Silylcyclopentenes via Ball Milling
Authors: Gao Pan , Li Yicheng , Zhang Duo , Zhang Guodong , Gao Liuzhou , Chen Feng
Published: June 2024

The development of innovative methods for synthesizing silylcyclopentene compounds is particularly important for enriching and improving the synthetical toolbox of organosilicon compounds. Herein, a facile approach has been developed for the synthesis of silylcyclopentenes promoted by mechanochemically generated organolithium species as silicon nucleophiles under ball milling conditions, avoiding the requirement of large amounts of bulk solvent. This operationally simple method demonstrates good functional group compatibility, which provides a great opportunity for further exploration of the synthetic applications of silylcyclopentenes. Density functional theory calculations indicated that the transient lithiosilole intermediates undergo a stepwise nucleophilic addition process, which governs this mechanic‐force‐promoted [4+1] cycloaddition reaction.
In Situ Generation and High Bioresistance of Trityl‐based Semiquinone Methide Radicals Under Anaerobic Conditions in Cellular Systems
Authors: Li Shuai , Deng Peng , Chang Qi , Feng Meirong , Shang Yixuan , Song Yuguang , Liu Yangping
Published: June 2024

Bioreduction of spin labels and polarizing agents (generally stable radicals) has been an obstacle limiting the in‐cell applications of pulsed electron paramagnetic resonance (EPR) spectroscopy and dynamic nuclear polarization (DNP). In this work, we have demonstrated that two semiquinone methide radicals (OXQM⋅ and CTQM⋅) can be easily produced from the trityl‐based quinone methides (OXQM and CTQM) via reduction by various reducing agents including biothiols and ascorbate under anaerobic conditions. Both radicals have relatively low pKa's and exhibit EPR single line signals at physiological pH. Moreover, the bioreduction of OXQM in three cell lysates enables quantitative generation of OXQM⋅ which was most likely mediated by flavoenzymes. Importantly, the resulting OXQM⋅ exhibited extremely high stability in the E.coli lysate under anaerobic conditions with 76‐ and 14.3‐fold slower decay kinetics as compared to the trityl OX063 and a gem‐diethyl pyrrolidine nitroxide, respectively. Intracellular delivery of OXQM into HeLa cells was also achieved by covalent conjugation with a cell‐permeable peptide as evidenced by the stable intracellular EPR signal from the OXQM⋅ moiety. Owing to extremely high resistance of OXQM⋅ towards bioreduction, OXQM and its derivatives show great application potential in in‐cell EPR and in‐cell DNP studies for various cells which can endure short‐term anoxic treatments.
Preparation of Dearomatized p‐Coumaric Acid Derivatives as DNA Damage Response Inhibitors with Potent In Vitro Antitumor Effect
Authors: Fási Laura , Gonda Tímea , Tóth Noémi , Vass Máté , Gyovai András , Nagy Viktória , Ocsovszki Imre , Zupkó István , Kúsz Norbert , Nové Márta , Spengler Gabriella , Berkecz Róbert , Wang Hui‐Chun , Chang Fang‐Rong , Hunyadi Attila
Published: June 2024

Our research group previously identified graviquinone (1) as a promising antitumor metabolite that is formed in situ when the antioxidant methyl caffeate scavenges free radicals. Furthermore, it exerted a DNA damaging effect on cancer cells and a DNA protective effect on normal keratinocytes. To expand and explore chemical space around qraviquinone, in the current work we synthesized 9 new alkyl‐substituted derivatives and tested their in vitro antitumor potential. All new compounds bypassed ABCB1‐mediated multidrug resistance and showed highly different cell line specificity compared with 1. All compounds were more potent in MDA‐MB‐231 than on MCF‐7 cells. The n‐butyl‐substituted derivatives 2 and 8 modulated the cell cycle and inhibited the ATR‐mediated phosphorylation of checkpoint kinase‐1 in MCF‐7 cells. As a significant expansion of our previous findings, our results highlight the potential antitumor value of alkyl‐substituted graviquinone derivatives.
Structure‐Metabolism Relationships of Benzimidazole Derivatives with anti‐Trypanosoma cruzi Activity for Chagas Disease
Authors: Espinoza‐Chávez Rocío Marisol , de Oliveira Rezende Júnior Celso , Laureano de Souza Mariana , Consolin Chelucci Rafael , Michelan‐Duarte Simone , Krogh Renata , Gomes Ferreira Leonardo Luiz , Valli Marilia , Sena de Oliveira Aldo , Andricopulo Adriano D. , Carlos Dias Luiz
Published: June 2024

This study introduces further insights from the hit‐to‐lead optimization process involving a series of benzimidazole derivatives acting as inhibitors of the cruzain enzyme, which targets Trypanosoma cruzi, the causative parasite of Chagas disease. Here, we present the design, synthesis and biological evaluation of 30 new compounds as a third generation of benzimidazole analogues with trypanocidal activity, aiming to enhance our understanding of their pharmacokinetic profiles and establish a structure‐metabolism relationships within the series. The design of these new analogues was guided by the analysis of previous pharmacokinetic results, considering identified metabolic sites and biotransformation studies. This optimization resulted in the discovery of two compounds (42 e and 49 b) exhibiting enhanced metabolic stability, anti‐Trypanosoma cruzi activity compared to benznidazole (the reference drug for Chagas disease), as well as being non‐cruzain inhibitors, and demonstrating a satisfactory in vitro pharmacokinetic profile. These findings unveil a new subclass of aminobenzimidazole and rigid compounds, which offer potential for further exploration in the quest for discovering novel classes of antichagasic compounds.
Development of New Peptidomimetic NADPH Oxidase Inhibitors with Antithrombotic Properties
Authors: Scalia Elisabetta , Chirco Antony , Calugi Lorenzo , Lenci Elena , Pagano Patrick J. , Pula Giordano , Trabocchi Andrea
Published: June 2024

The ability of synthetic peptides inhibitors of NOX1 to effectively block the production of ROS by the enzyme was studied with different methodologies. Specifically, taking advantage of our understanding of the active epitope of the regulatory NOX1 subunit NOXA1 as a potent inhibitor of NOX1‐derived O2⋅− formation, a panel of peptidomimetic derivatives of this peptide were designed and synthesized with the aim of improving their activity and increasing their stability in plasma. The results highlighted that improved efficacy and potency was found for both the peptide‐peptoid hybrid GS2, whereas stapled peptide AC5 and its precursor showed higher stability despite lower biological potency. This study showed that minimal structural modifications of NOXA1 peptides are required to improve both their potency and stability to finally achieve best candidates as new potential anti‐thrombotic drugs.
Comparative Bioequivalence Study of 2 Clopidogrel 75‐mg Tablet Formulations in Moroccan Volunteers
Authors: Orche Aimen El , Khabbaz Choukri El , Cheikh Amine , Bouchafra Houda , Jawhari Samira , Abbes Faouzi My El , Cherrah Yahya
Published: June 2024

This study investigates the pharmacokinetic properties and bioequivalence of 2 formulations of clopidogrel tablets administered to a cohort of healthy Moroccan male volunteers. The primary objective was to assess the rate and extent of drug absorption from the test formulation in comparison to a reference formulation, focusing on critical parameters including maximum plasma concentration (Cmax), area under the concentration‐time curve from 0 to the last measurable time (AUC0‐t), and area under the concentration‐time curve extrapolated to infinity (AUC0‐∞). The results revealed that the geometric mean ratios of Cmax, AUC0‐t, and AUC0–∞ for the test formulation relative to the reference formulation were 105.7%, 105.6%, and 105.6%, respectively. The 90% confidence intervals for these parameters fell within the predefined bioequivalence range of 80%‐125%, indicating a statistically and clinically equivalent performance between the 2 formulations. This investigation sheds light on the pharmacokinetic behavior of clopidogrel in the context of the Moroccan male population, offering valuable insights into the comparability of formulations.
Structural Order in the Hydration Shell of Nonpolar Groups versus that in Bulk Water
Authors: Graziano Giuseppe
Published: June 2024

The poor solubility of nonpolar compounds in water around room temperature is governed by a large and negative entropy change, whose molecular cause is still debated. Since the Frank and Evans original proposal in 1945, the large and negative entropy change is usually attributed to the formation of ordered structures in the hydration shell of nonpolar groups. However, the existence of such ordered structures has never been proven. The present study is aimed at providing available structural results and thermodynamic arguments disproving the existence of ordered structures in the hydration shell of nonpolar groups.
Construction of BiVO4@ NiCo2O3 Heterojunction to Promote Photocatalytic CO2 Reduction
Authors: Fu Wenjie , Lu Guanhong , Wang Xiao , Xie Xiaofeng , Sun Jing
Published: June 2024

Constructing a catalyst capable of reducing CO2 through photoreduction in aqueous environments presents a significant challenge. In this study, we present the synthesis of BiVO4@NiCo2O3 heterojunction using a straightforward hydrothermal method for CO2 photoreduction. The sample with the optimal loading ratio demonstrates a CO generation rate of 7.202 μmol ⋅ g−1 ⋅ h−1, which is twice that of pure BiVO4 (3.626 μmol ⋅ g−1 ⋅ h−1) and 1.5 times that of pure NiCo2O3 (4.726 μmol ⋅ g−1 ⋅ h−1). Analysis using XPS and EPR techniques suggests that electron transfer at the interface of the heterojunction facilitates the separation of photogenerated charge carriers, thereby enhancing the efficiency of the photocatalytic process. This investigation offers a viable approach for developing photocatalysts for CO2 reduction in aqueous environments.
Antimicrobial Efficacy of Photocaged β‐Lapachone in Bacillus subtilis Biofilms
Authors: Hudson Elyse , Faylinn Christabel , Lopez‐Miranda Ivonne R. , Milstein Joshua N. , Beharry Andrew A.
Published: June 2024

With the rise of antibiotic resistance within clinical settings, combating the growth of microbial biofilms presents a unique challenge. Biofilm‐inhabiting bacteria are embedded within a self‐produced, protective matrix, which can reduce the efficacy of treatment. The naturally derived product β‐lapachone is an appealing therapeutic agent that has been reported to inhibit biofilm growth. However, its off‐target toxicity and poor metabolic stability pose a significant hurdle for its application in vivo. Using a photo‐pharmacological approach via a coumarin‐based photocage, the reactivity of β‐lapachone can be tuned so it only becomes active once the photocage is removed. Here we report both the photo‐uncaging efficiency and the effective inhibition concentration of photocaged β‐lapachone within model Bacillus subtilis biofilms. Additionally, the mechanism of action is analyzed with results supporting catalase inhibition. This novel light‐activatable anti‐microbial has potential applications in medical settings to inhibit biofilm growth and provide synergistic treatment with traditional antibiotics.
Performing Suction Blister Skin Biopsies
Authors: MacDonald Elizabeth A. , Katz Erica L. , Pearson Todd F. , Harris John E.
Published: June 2024

Traditional skin sampling methods include punch or shave biopsies to produce a solid tissue sample for analysis. These biopsy procedures are painful, require anesthesia, and leave permanent scars. This unit describes a suction blister skin biopsy method that can be used in place of traditional biopsy methodologies as a minimally invasive, non‐scarring skin sampling technique. The induction of suction blisters uses an instrument with a chamber that applies negative pressure and gentle heat to the skin. Blister formation occurs within 1 hr, producing up to five blisters, each 10 mm in diameter per biopsy site. Blister fluid can be extracted and centrifuged to retrieve cells from the epidermis and upper dermis for flow cytometry, single‐cell RNA sequencing, cell culture, and more without the need for digestion protocols. In addition, the blister fluid can be used to measure soluble proteins and metabolites. This unit describes the preparation of supplies and subjects, the suction blister biopsy procedure and blister formation, fluid extraction, and post‐blistering care. © 2024 Wiley Periodicals LLC.

Basic Protocol 1: Preparation of supplies and subject

Basic Protocol 2: Suction blister biopsy procedure and formation

Basic Protocol 3: Blister fluid extraction

Basic Protocol 4: Post‐blister care and clean up
P‐NiFe2O4/N‐Doped Carbon Nanotubes/NiFe Multi‐Phase Heterojunctions for Overall Water Splitting and Urea Electrolysis
Authors: Jia Lina , Du Gaohui , Han Di , Wang Yunting , Wang Youqing , Li Huayu , Zhao Wenqi , Chen Shixian , Zhang Miao , Su Qingmei , Xu Bingshe
Published: June 2024

The design and construction of highly efficient electrocatalysts for overall water splitting and urea electrolysis are significantly important for promoting energy conversion and realizing green hydrogen production. In this work, we constructed a multi‐phase heterojunction through a simple hydrothermal and phosphorization process. The P‐doped NiFe2O4 (P‐NiFe2O4) nanoparticles were uniformly anchored on the bamboo‐like N‐doped carbon nanotubes (NCNTs) grown via a NiFe‐alloy autocatalysis. The electronic structure and coordination environment of active species were optimized by the synergistic action of P doping, well‐dispersed ultrafine NiFe2O4, and NCNTs matrix with good conductivity, enhancing their quantity and activity for electrocatalysis. Consequently, the P‐NiFe2O4/NCNTs/NiFe exhibits excellent HER and OER activities with an overpotential of 111 and 266 mV at 10 mA cm−2 in 1 M KOH, respectively. The symmetrical overall water‐splitting cell using P‐NiFe2O4/NCNTs/NiFe as both anode and cathode delivers 10 mA cm−2 at a voltage of 1.604 V in 1 M KOH. Notably, the two‐electrode cell requires a low voltage of 1.467 V to achieve a current density of 10 mA cm−2 in 1 M KOH solution with 0.6 M urea. This designed catalysts display outstanding reaction kinetics and catalytic stability. This work provides useful guidance for applying transition metal‐based catalysts for hydrogen production.
Tear exosome‐based PROteolysis TArgeting Chimeras nanomedicine for human immunodeficiency virus‐mediated cancer treatment
Authors: Mukerjee Nobendu
Published: June 2024

Human Immunodeficiency Virus (HIV) significantly increases the risk of various cancers due to chronic immune suppression and viral oncogenes. Traditional therapies, including antiretroviral therapy (ART), chemotherapy, and radiation, often face limitations such as drug resistance and systemic toxicity. PROteolysis TArgeting Chimeras (PROTACs) have emerged as a promising approach for targeted protein degradation, offering significant advantages over conventional treatments. However, effective delivery remains a challenge. This paper explores the innovative use of tear exosome‐based delivery systems for PROTACs in treating HIV‐mediated cancers. Tear exosomes, due to their natural origin, biocompatibility, and inherent targeting capabilities, present a novel and effective platform for delivering PROTACs, enhancing therapeutic specificity and reducing adverse effects. Integrating the unique properties of tear exosomes with the therapeutic potential of PROTACs could revolutionize the treatment of HIV‐mediated cancers by overcoming current therapeutic challenges and improving patient outcomes.
Chloride Modulates Central pH Sensitivity and Plasticity of Brainstem Breathing‐Related Biorhythms in Zebra Finch Embryos
Authors: Whitaker‐Fornek Jessica R. , Nelson Jennie K. , Pilarski Jason Q.
Published: June 2024

All terrestrial vertebrate life must transition from aquatic gas exchange in the embryonic environment to aerial or pulmonary respiration at birth. In addition to being able to breathe air, neonates must possess functional sensory feedback systems for maintaining acid–base balance. Respiratory neurons in the brainstem act as pH sensors that can adjust breathing to regulate systemic pH. The central pH sensitivity of breathing‐related motor output develops over the embryonic period in the zebra finch (Taeniopygia guttata). Due to the key role of chloride ions in electrochemical stability and developmental plasticity, we tested chloride's role in the development of central pH sensitivity. We blocked gamma‐aminobutyric acid‐A receptors and cation‐chloride cotransport that subtly modulated the low‐pH effects on early breathing biorhythms. Further, chloride‐free artificial cerebrospinal fluid altered the pattern and timing of breathing biorhythms and blocked the stimulating effect of acidosis in E12–14 brainstems. Early and middle stage embryos exhibited rebound plasticity in brainstem motor outputs during low‐pH treatment, which was eliminated by chloride‐free solution. Results show that chloride modulates low‐pH sensitivity and rebound plasticity in the zebra finch embryonic brainstem, but work is needed to determine the cellular and circuit mechanisms that control functional chloride balance during acid–base disturbances.
Disturbance‐mediated changes to boreal mammal spatial networks in industrializing landscapes
Authors: Curveira‐Santos Gonçalo , Marion Solène , Sutherland Chris , Beirne Christopher , Herdman Emily J. , Tattersall Erin R. , Burgar Joanna M. , Fisher Jason T. , Burton A. Cole
Published: June 2024

Compound effects of anthropogenic disturbances on wildlife emerge through a complex network of direct responses and species interactions. Land‐use changes driven by energy and forestry industries are known to disrupt predator–prey dynamics in boreal ecosystems, yet how these disturbance effects propagate across mammal communities remains uncertain. Using structural equation modeling, we tested disturbance‐mediated pathways governing the spatial structure of multipredator multiprey boreal mammal networks across a landscape‐scale disturbance gradient within Canada's Athabasca oil sands region. Linear disturbances had pervasive direct effects, increasing site use for all focal species, except black bears and threatened caribou, in at least one landscape. Conversely, block (polygonal) disturbance effects were negative but less common. Indirect disturbance effects were widespread and mediated by caribou avoidance of wolves, tracking of primary prey by subordinate predators, and intraguild dependencies among predators and large prey. Context‐dependent responses to linear disturbances were most common among prey and within the landscape with intermediate disturbance. Our research suggests that industrial disturbances directly affect a suite of boreal mammals by altering forage availability and movement, leading to indirect effects across a range of interacting predators and prey, including the keystone snowshoe hare. The complexity of network‐level direct and indirect disturbance effects reinforces calls for increased investment in addressing habitat degradation as the root cause of threatened species declines and broader ecosystem change.
A framework for identifying factors controlling cyanobacterium Microcystis flos‐aquae blooms by coupled CCM–ECCM Bayesian networks
Authors: Tal O. , Ostrovsky I. , Gal G.
Published: June 2024

Cyanobacterial blooms in freshwater sources are a global concern, and gaining insight into their causes is crucial for effective resource management and control. In this study, we present a novel computational framework for the causal analysis of cyanobacterial harmful algal blooms (cyanoHABs) in Lake Kinneret. Our framework integrates Convergent Cross Mapping (CCM) and Extended CCM (ECCM) causal networks with Bayesian Network (BN) models. The constructed CCM–ECCM causal networks and BN models unveil significant interactions among factors influencing cyanoHAB formation. These interactions have been validated by domain experts and supported by evidence from peer‐reviewed publications. Our findings suggest that Microcystis flos‐aquae levels are influenced not only by community structure but also by ammonium, phosphate, oxygen, and temperature levels in the weeks preceding bloom occurrences. We demonstrated a non‐parametric computational framework for causal analysis of a multivariate ecosystem. Our framework offers a more comprehensive understanding of the underlying mechanisms driving M. flos‐aquae blooms in Lake Kinneret. It captures complex interactions and provides an explainable prediction model. By considering causal relationships, temporal dynamics, and joint probabilities of environmental factors, the proposed framework enhances our understanding of cyanoHABs in Lake Kinneret.
Warming effects on the life cycles of two parasitic copepods with different invasion histories
Authors: Jolma Elli Rosa Emilia , Born‐Torrijos Ana , Heesterbeek Hans , van Leeuwen Anieke , van Leeuwen Sonja Maria , Twijnstra Robert H. , Wegner K. Mathias , Thieltges David W.
Published: June 2024

Climate change may exacerbate the impact of invasive parasites from warmer climates through pre‐existing temperature adaptations. We investigated temperature impacts on two closely related marine parasitic copepod species that share the blue mussel (Mytilus edulis) as host: Mytilicola orientalis has invaded the system from a warmer climate <20 years ago, whereas its established congener Mytilicola intestinalis has had >90 years to adapt. In laboratory experiments with temperatures 10–26°C, covering current and future temperatures as well as heat waves, the development of both life cycle stages of both species accelerated with increasing temperature. In the parasitic stages, the growth of the established invader increased evenly from 10°C to 22°C, whereas the recent invader barely grew at all at 10°C and grew faster already at 18°C. In contrast, temperature had little effect on the transition success between life cycle stages. However, the highest temperature (26°C) limited the egg development success of the established invader and the host entry success of both species, whereas the infection success of the established invader increased at 18°C and 22°C. In general, our experiments indicate that the main effect of temperature on both species is through development speed and not life cycle stage transition success. Based on regional long‐term temperature data and predictions, the numbers of completed life cycles per year will increase for both parasites. The established invader seems better adapted for low current temperatures (around 10°C), whereas the more recent invader barely develops at these temperatures but can cope in high temperatures (around 26°C). Hence, pre‐existing temperature adaptations of the recent invader may allow the species to better cope with heat waves.
Dispersal limitation and environmental filtering effects: The taxonomic and functional beta diversity of ground beetles along the altitudinal gradient in Chinese warm‐temperature forests
Authors: Shen Yagang , Zou Yi , Song Kun , Wan Xia
Published: June 2024

Beta diversity patterns along environmental gradients and underlying mechanisms constitute key research inquiries in biogeography. However, ecological processes often also influence the functional traits of biological communities, making the assessment of functional β‐diversity crucial. Ground beetles (Coleoptera: Carabidae) are one of the most species‐rich groups in the insect community, displaying strong habitat specificity and morphological differences. In this study, we explored the patterns of taxonomic and functional beta diversity in ground beetle communities along the altitudinal gradient of warm‐temperature forests. By partitioning beta diversity into turnover and nestedness components, we evaluated their relationship with spatial distance. Our findings indicate a decline in species and functional trait similarity with increasing elevation and geographic distance. Further analysis attributed both types of beta diversity in carabids to a combination of dispersal limitation and environmental filtering, with elevation and geographic distance emerging as significant factors. Interestingly, forest‐type variations were found to have no impact on the beta diversity of these communities. Our study reveals the impact of environmental filtering and dispersal limitation on both taxonomic and functional beta‐diversity, shedding light on carabid community assembly in localized warm‐temperature forest areas in eastern China.
Designing eco‐evolutionary experiments for restoration projects: Opportunities and constraints revealed during stickleback introductions
Authors: Hendry Andrew P. , Barrett Rowan D. H. , Bell Alison M. , Bell Michael A. , Bolnick Daniel I. , Gotanda Kiyoko M. , Haines Grant E. , Lind Åsa J. , Packer Michelle , Peichel Catherine L. , Peterson Christopher R. , Poore Hilary A. , Massengill Robert L. , Milligan‐McClellan Kathryn , Steinel Natalie C. , Sanderson Sarah , Walsh Matthew R. , Weber Jesse N. , Derry Alison M.
Published: June 2024

Eco‐evolutionary experiments are typically conducted in semi‐unnatural controlled settings, such as mesocosms; yet inferences about how evolution and ecology interact in the real world would surely benefit from experiments in natural uncontrolled settings. Opportunities for such experiments are rare but do arise in the context of restoration ecology—where different “types” of a given species can be introduced into different “replicate” locations. Designing such experiments requires wrestling with consequential questions. (Q1) Which specific “types” of a focal species should be introduced to the restoration location? (Q2) How many sources of each type should be used—and should they be mixed together? (Q3) Which specific source populations should be used? (Q4) Which type(s) or population(s) should be introduced into which restoration sites? We recently grappled with these questions when designing an eco‐evolutionary experiment with threespine stickleback (Gasterosteus aculeatus) introduced into nine small lakes and ponds on the Kenai Peninsula in Alaska that required restoration. After considering the options at length, we decided to use benthic versus limnetic ecotypes (Q1) to create a mixed group of colonists from four source populations of each ecotype (Q2), where ecotypes were identified based on trophic morphology (Q3), and were then introduced into nine restoration lakes scaled by lake size (Q4). We hope that outlining the alternatives and resulting choices will make the rationales clear for future studies leveraging our experiment, while also proving useful for investigators considering similar experiments in the future.
Effects of dietary nutrients of the gut microbiota in the long‐tailed dwarf hamster (Cricetulus longicaudatus)
Authors: Cao Kanglin , Tao Mengfan , Pu Xinsheng , Hou Yu , Ren Yue , Liu Wei , Yang Xin'gen
Published: June 2024

Gut microbiota is a key factor in maintaining the dietary and metabolic homeostasis of small mammals. To explore the effect of diet on the gut microbiota of the long‐tailed dwarf hamster (Cricetulus longicaudatus), 16S rDNA high‐throughput sequencing combined with bioinformatics analysis was used to investigate the succession process of the gut microbiota and effects of different nutrients on the composition and function of the gut microbiota. The results showed that diet structure can significantly influence the composition and function of the gut microbiota, as well as the health of animals. The highest relative abundance of Firmicutes, and the simplest co‐occurrence network occurred in the wild. Whereas the relative abundance of Bacteroidetes is higher and the most complex network structure was observed after 35 days of same feeding. Compared to the other four groups, the relative abundance of Firmicutes in the wheat + peanuts (WP) group was the highest after 35 days of different feeding, and the highest relative abundance of Bacteroidetes occurred in the wheat‐only (WH) group. Bacteroidetes exhibit carbohydrate degradation activity, and Firmicutes are strongly associated with fat uptake. We also found a significant positive correlation between Lactobacillus and body weight, indicating that Lactobacillus plays a crucial role in modulating fat intake and weight management. This study provides empirical evidence to facilitate the understanding of the co‐evolutionary dynamics between C. longicaudatus and their gut microbiota and establishes a theoretical foundation for utilizing gut microbiota in rodent control.
Comparative mitogenome research revealed the phylogenetics and evolution of the superfamily Tenebrionoidea (Coleoptera: Polyphage)
Authors: Hu Yun‐Jian , Jia Feng‐Fan , Hu Li , Wu Chuan , Tian Tian , Li Ting‐Jing , Chen Bin
Published: June 2024

Despite the worldwide distribution and rich diversity of the superfamily Tenebrionoidea, the knowledge of the mitochondrial genomes (mtgenome) characteristics of the superfamily is still very limited, and its phylogenetics and evolution remain unresolved. In the present study, we newly sequenced mtgenomes from 19 species belonging to Tenebrionoidea, and a total of 90 mitochondrial genomes from 16 families of Tenebrionoidea were used for phylogenetic analysis. There exist 37 genes for all 82 species of complete mtgenomes of 16 families investigated, and their characteristics are identical as reported mtgenomes of other Tenebrionoids. The Ka/Ks analysis suggests that all 13 PCGs have undergone a strong purifying selection. The phylogenetic analysis suggests the monophyly of Mordellidae, Meloidae, Oedemeridae, Pyrochroidae, Salpingidae, Scraptiidae, Lagriidae, and Tenebrionidae, and the Mordellidae is close to the Ripiphoridae. The “Tenebrionidae clade” and “Meloidae clade” are monophyletic, and both of them are sister groups. In the “Meloidae clade,” Meloidae is close to Anthicidae. In the “Tenebrionidae clade,” the family Lagriidae and Tenebrionidae are sister groups. The divergence time analysis suggests that Tenebrionoidea originated in the late Jurassic, Meloidae Mordellidae, Lagriidae, and Tenebrionidae in the Cretaceous, Oedemeridae in Paleogene. The work lays a base for the study of mtgenome, phylogenetics, and evolution of the superfamily Tenebrionoidea.
Invasive‐plant traits, native‐plant traits, and their divergences as invasion factors
Authors: Li Yang , Yue Ming , Wang Yuchao , Mao Zhuxin , Lyv Jinlin , Li Qian
Published: June 2024

Invasive plants exert significant ecological impacts on native plants, communities, and ecosystems. However, consistent conclusions regarding how traits of invasive plants, native plants, and their divergences affect invasion dynamics are still lacking. Here, we conducted a pairwise common garden experiment to investigate how invasion was influenced not only by invasive plants but also by native plants, aiming to elucidate the role of invasive‐plant traits, native‐plant traits, and their divergences in invasion processes. Our findings revealed variations in invasive stage depending on the combinations of invasive and native plants. Specifically, native plants such as A. argyi, A. lavandulifolia, and C. album exhibited competitive superiority when co‐occurring with the three invasive plants. S. viridis, A. vestita, and A. annua had competitive superiority when they co‐occurred with E. canadensis, G. quadriradiata, and E. annuus respectively. Furthermore, our results demonstrated that the competitive abilities of invasive plants were primarily influenced by factors such as height, diameter, and biomass allocation, while native plants' competitive abilities were mainly affected by diameter, biomass allocation, and function group differences. Moreover, our analysis revealed that invasive‐plant traits, native‐plant traits, their divergences, and their interactions together explained 36.88% of the variation in invasion dynamics, with invasive‐plant traits and the native‐plant traits explaining 10.19% and 6.88%, respectively. In conclusion, the traits of invasive and native plants, along with their divergences, significantly influence interspecific relationships, and influencing the invasive stages. Divergences in competitive strategies between the native plants and invasive plants facilitated invasion processes. Our study not only contributes to understanding the mechanisms underlying invasion, but also provides a scientific foundation for predicting and managing the negative effects of invasive plants.