bluuebunny/crossref_metadata_2025 huggingface dataset:
SHARD: an improved method for staining and visualizing multiplex immunofluorescence in optically cleared postmortem human brain tissue
Authors: Rosen Grace A. , Kirsch Daniel , Nicks Raymond , Kelley Hunter , Mathias Rebecca , Cormier Kerry A. , Kubilus Caroline A. , Dec Bryan , Stein Thor D. , Alvarez Victor E. , Alosco Michael L. , McKee Ann C. , Huber Bertrand R.
Published: October 2024

Postmortem human brain tissue is a critical resource for studying neurodegenerative disease, providing critical insights into cellular morphology, pathology, and network connectivity. To improve standard microscopy and enable high-resolution, three-dimensional (3D) images of tissues at the subcellular level, tissue-clearing methods have been developed. These 3D images allow for the analysis of large regions of interest and can be used to study structural and spatial changes that occur during neurodegeneration. Additionally, 3D imaging facilitates the visualization of whole-cell morphology, especially in cells with long processes that would otherwise be truncated in single-plane images. Human brain tissue is especially challenging for tissue clearing due to the abundance of lipids in myelin and the need for optimal fixation and low postmortem intervals. Formaldehyde-based fixatives, commonly used in preserving tissue, hinder antibody binding by crosslinking important antibody epitopes, and fluorescent microscopy requires the incorporation of fluorescent labels through passive diffusion or electrophoresis. Recent studies have focused on optimally fixed human brain tissue with short postmortem intervals, limiting the general applicability of these methods. To address these challenges, we developed SHARD (SHIELD, antigen retrieval, and delipidation), a simple and widely applicable method for clearing and labeling human brain tissue, which can be applied to long-term banked human brain tissue preserved in formaldehyde. SHARD is a novel addition to the SHIELD tissue clarification method, combining antigen retrieval, tissue clearing, and staining of 200-μm sections from long-term banked human brain tissue. The SHARD method is effective for postmortem intervals (PMIs) ranging from 10 to 72 h in multiple neurodegenerative diseases and control samples. In this study, we demonstrate that the SHARD method significantly enhances the immunostaining of glial fibrillary acidic protein (GFAP), an astrocytic cytoskeletal marker. Overall, the combination of antigen retrieval and tissue delipidation holds great potential for achieving detailed 3D immunostaining in long-term formaldehyde-fixed postmortem human brain tissue, opening new avenues for research and discovery.
Effects of plant-based diets combined with exercise training on leptin and adiponectin levels in adults with or without chronic diseases: a systematic review and meta-analysis of clinical studies
Authors: Kazeminasab Fatemeh , Fatemi Rouholah , Bagheri Reza , Santos Heitor O. , Dutheil Fred
Published: October 2024

Background

The effects of exercise training combined with plant-based diets (PBD) on leptin and adiponectin levels have been studied. However, little is known regarding the impact of exercise training combined with PBD on leptin and adiponectin levels in adults with or without chronic diseases.

Methods

PubMed, Web of Science, and Scopus were searched to identify original articles, published until May 2024, to assess the effects of exercise training combined with PBD on leptin and adiponectin levels in adults with or without chronic diseases. Standardized mean differences (SMD) and 95% confidence intervals were calculated using random models.

Results

Nine studies comprising 960 participants with overweight and obesity were included in the current meta-analysis. Exercise training combined with PBD reduced leptin [SMD = -0.33 (95% CI: −0.62 to −0.04); p = 0.025] while increasing adiponectin [SMD = 0.93 (95% CI: 0.12 to 1.74); p = 0.024] levels.

Conclusion

Exercise training combined with PBD is suggested as a non-invasive intervention for reducing leptin while increasing adiponectin levels to control body mass and other disorders related to obesity in adults.
Breast and axillary marking in the neoadjuvant setting: survey results from experts of the Brazilian society of mastology
Authors: Couto Henrique Lima , Hassan Augusto Tufi , Steinmacher Dalton Ivan , Pessoa Eduardo Carvalho , Millen Eduardo Camargo , Zerwes Felipe , Cavalcante Francisco Pimentel , Tosello Giuliano , Novita Guilherme , Machado Badan Gustavo , Esteves Francisco José Luis , Soares Leonardo Ribeiro , Budel Lucas Roskamp , Fernandes Chala Luciano , Fernandes Raquel Civolani Marques , Freitas-Junior Ruffo , Oliveira Vilmar Marques de , Budel Vinicius Milani , Mattar André
Published: October 2024

Introduction/objectives

The precise location of the tumor site is essential for the success of surgical treatment. Neoadjuvant chemotherapy (NAC) is a challenge for preoperative tumor and node localization. Thus, the knowledge and attitudes of the affiliated members of the Brazilian Society of Mastology (SBM) regarding breast and axilla marking were evaluated and a consensus regarding management and treatment was reached.

Methods

This was an online survey conducted between June and December 2022. All 1,742 active mastologists affiliated to the SBM were invited anonymously. The online form contained 28 objective questions, of which 22 were formulated on a Likert scale. These questions addressed relevant aspects related to breast and axilla marking in the neoadjuvant setting. Responses that reached 70% agreement were considered consensual. Statistical analysis was performed using the SPSS program version 26.0. Post hoc analysis was performed when appropriate and the significance level was set at p < 0.05. Polychoric regression analyses were conducted using `VGAM` package

Results

In total, 468 mastologists answered the questionnaire (26.8%), with a predominance of professionals aged between 40–49 years (32.1%). Most professionals were board-certified (84,8%). The indication of tumor marking in the breast prior to NAC was consensual (96.4%) and the metal clip was the preferred method (69.7%). There was no consensus regarding the indication of pre-NAC histologically positive lymph node marking (49.8% disagree and 42.8% agree). However, there was consensus that the clinical and imaging evaluation was insufficient for staging the axilla as N1 (71.6%). The contraindication of breast and node marking in T4b tumors (71.2%) was consensual. There was consensus on the indication of sentinel lymph node biopsy (SLNB) for initially cN1 (92.3%) or cN2 (72.7%) tumors that became cN0 after NAC, with 67.5% opting for dual staining with technetium and patent blue. When <3 lymph nodes were retrieved 41.0% of mastologists performed axillary lymphadenectomy. Among the 28 questions, consensus was reached on only 11 (39.3%).

Conclusion

The indication of pre-NAC breast marking is consensual among Brazilian mastologists, although axillary nodal marking is not. There is a great divergence of attitudes among Brazilian surgeons in relation to the many issues related to pre-NAC breast and axilla marking.
Predictive value of methylene blue combined with indocyanine green in sentinel lymph node metastasis in breast cancer: a prospective pilot cohort study
Authors: He Zecheng , Guo Fan , Liu Yuhan , Lin Yan , Wang Changjun , Zhou Yidong , Sun Qiang
Published: October 2024

Background

The status of sentinel lymph nodes is crucial for prognosis and treatment decisions in breast cancer patients. This study aimed to evaluate the predictive value of combined methylene blue and indocyanine green for sentinel lymph node metastasis in breast cancer.

Methods

This prospective cohort study enrolled 90 clinically node-negative breast cancer patients. Methylene blue and indocyanine green were injected locally before surgery. Sentinel lymph nodes were grouped based on fluorescence intensity and methylene blue staining. A binary logistic regression model was established using 285 lymph node groups to predict metastatic risk.

Results

A total of 475 lymph nodes were identified, with 33 being metastatic. The metastatic risk reached 70% for partially blue-stained and weakly fluorescent lymph nodes between 1-2 cm. The model revealed associations between lymph node size, dye staining patterns, and metastatic risks (P<0.05). The AUC of the ROC curve was 0.855.

Conclusions

The staining pattern of combined methylene blue and indocyanine green could predict risks of sentinel lymph node metastasis and facilitate rapid intraoperative identification of high-risk lymph nodes.
Prediction of benign and malignant ground glass pulmonary nodules based on multi-feature fusion of attention mechanism
Authors: Deng Heng , Huang Wenjun , Zhou Xiuxiu , Zhou Taohu , Fan Li , Liu Shiyuan
Published: October 2024

Objectives

The purpose of this study was to develop and validate a new feature fusion algorithm to improve the classification performance of benign and malignant ground-glass nodules (GGNs) based on deep learning.

Methods

We retrospectively collected 385 cases of GGNs confirmed by surgical pathology from three hospitals. We utilized 239 GGNs from Hospital 1 as the training and internal validation set, and 115 and 31 GGNs from Hospital 2 and Hospital 3, respectively, as external test sets 1 and 2. Among these GGNs, 172 were benign and 203 were malignant. First, we evaluated clinical and morphological features of GGNs at baseline chest CT and simultaneously extracted whole-lung radiomics features. Then, deep convolutional neural networks (CNNs) and backpropagation neural networks (BPNNs) were applied to extract deep features from whole-lung CT images, clinical, morphological features, and whole-lung radiomics features separately. Finally, we integrated these four types of deep features using an attention mechanism. Multiple metrics were employed to evaluate the predictive performance of the model.

Results

The deep learning model integrating clinical, morphological, radiomics and whole lung CT image features with attention mechanism (CMRI-AM) achieved the best performance, with area under the curve (AUC) values of 0.941 (95% CI: 0.898-0.972), 0.861 (95% CI: 0.823-0.882), and 0.906 (95% CI: 0.878-0.932) on the internal validation set, external test set 1, and external test set 2, respectively. The AUC differences between the CMRI-AM model and other feature combination models were statistically significant in all three groups (all p<0.05).

Conclusion

Our experimental results demonstrated that (1) applying attention mechanism to fuse whole-lung CT images, radiomics features, clinical, and morphological features is feasible, (2) clinical, morphological, and radiomics features provide supplementary information for the classification of benign and malignant GGNs based on CT images, and (3) utilizing baseline whole-lung CT features to predict the benign and malignant of GGNs is an effective method. Therefore, optimizing the fusion of baseline whole-lung CT features can effectively improve the classification performance of GGNs.
The characteristics of high-risk HPV-negative cervical cancer: a systematic review and meta-analysis
Authors: Lou Jicheng , Guo Feng
Published: October 2024

Introduction

In order to comprehensive understanding of the characteristics of high-risk HPV-negative cervical cancer and aiming to provide certain evidence in the diagnosis and treatment of this kind of disease, the clinical and pathological characteristics of high-risk HPV-negative (hr-HPV-negative) cervical cancer was evaluated through systematic review and meta-analysis.

Methods

We employed a systematic search strategy incorporating both MeSH terms and free words across multiple databases including PubMed, Embase, Cochrane Library, Web of Science, CNKI and Wanfang. The strategy was specifically designed to identify articles investigating risk factors or clinical characteristics of hr-HPV-negative cervical cancer. Our database search spanned from their inception until October 2023. The literature selection and data collection processes were guided by predetermined inclusion and exclusion criteria. Each included research was independently evaluated by two reviewers using the Newcastle-Ottawa Scale (NOS) and a meta-analysis was conducted utilizing RevMan5.4.

Results

In total, nine studies published between 2013 and 2020 were included. The risk of lymph node metastasis was notably higher in hr-HPV-negative cervical cancer patients compared to hr-HPV-positive counterparts [RR=1.99, 95%CI (1.46–2.72), P<0.0001]. It was found that clinical staging of Ib2 or higher grade was significantly increased in hr-HPV-negative cervical cancer patients, as reported by seven studies [RR=1.42, 95%CI (1.15–1.77), P=0.0001]. Non-squamous carcinoma incidence, reported in seven studies, was significantly higher in hr-HPV-negative patients [RR=2.56, 95%CI (1.63–4.01), P=0.001]. Adenocarcinoma incidence was also significantly higher in hr-HPV-negative cervical cancer patients [RR=2.36, 95%CI (1.44–3.89), P=0.005].

Conclusions

Cervical cancer patients not infected by hr-HPV exhibited increased rates of lymph node metastasis, adenocarcinoma, and non-squamous carcinoma. And they were often diagnosed at more advanced stages. These variables suggest that hr-HPV-negative cervical cancer patients presented with a more severe health status at the time of their initial cancer diagnosis.
From defense to offense: antimicrobial peptides as promising therapeutics for cancer
Authors: Zare-Zardini Hadi , Saberian Elham , Jenča Andrej , Ghanipour-Meybodi Razieh , Jenča Andrej , Petrášová Adriána , Jenčová Janka
Published: October 2024

Antimicrobial peptides (AMPs), naturally occurring components of innate immunity, are emerging as a promising new class of anticancer agents. This review explores the potential of AMPs as a novel class of anticancer agents. AMPs, naturally occurring peptides with broad-spectrum antimicrobial activity, exhibit several characteristics that make them attractive candidates for cancer therapy, including selectivity for cancer cells, broad-spectrum activity, and immunomodulatory effects. Analysis of a dataset of AMPs with anticancer activity reveals that their effectiveness is influenced by various structural properties, including net charge, length, Boman index, and hydrophobicity. These properties contribute to their ability to target and disrupt cancer cell membranes, interfere with intracellular processes, and modulate the immune response. The review highlights the promising potential of AMPs as a new frontier in cancer treatment, offering hope for more effective and less toxic therapies. AMPs demonstrate promising potential in cancer therapy through multiple mechanisms, including direct cytotoxicity, immune response modulation, and targeting of the tumor microenvironment, as evidenced by extensive preclinical studies in animal models showing tumor regression, metastasis inhibition, and improved survival rates. AMPs show significant potential as cancer therapeutics through their direct cytotoxicity, immune response modulation, and tumor microenvironment targeting, with promising results from preclinical studies and early-phase clinical trials. Future research should focus on optimizing AMP properties, developing novel delivery strategies, and exploring synergistic combination therapies to fully realize their potential as effective cancer treatments, while addressing challenges related to stability, delivery, and potential toxicity.
Case Report: Reduction in post-amputation phantom limb pain intensity accompanying the onset of phantom limb telescoping
Authors: Aternali Andrea , Lumsden-Ruegg Heather , Appel Lora , Hitzig Sander L. , Mayo Amanda L. , Katz Joel
Published: October 2024

Introduction

Individuals with limb loss frequently report post-amputation phenomena, including nonpainful phantom sensations, phantom limb pain (PLP), and residual limb pain (RLP). Although post-amputation pain is common, not all patients benefit from widely accepted treatments. A greater understanding of phantom limb “telescoping”, the experience of one's phantom hand or foot gradually approaching the residual limb, may assist in developing more effective interventions for reducing post-amputation pain. This case report explores the relationships between PLP, RLP, telescoping, and psychosocial experience in one person with a lower limb amputation. The aim of this case is to illustrate one possible relationship between telescoping and PLP as the mechanisms linking the two remain equivocal.

Methods

The participant is a 35-year-old male who underwent a transfemoral amputation due to a traumatic injury to his right leg approximately 4 years prior. He responded to questionnaires evaluating demographic and health-related information (e.g., age, sex, marital status, reason for amputation), pain and psychological variables via the Brief Pain Inventory (BPI-SF), ID Pain Questionnaire (IDPQ), Pain Catastrophizing Scale (PSC-4), Patient Health Questionnaire-4 (PHQ-4), Life Orientation Test-Revised (LOT-R), Connor-Davidson Resilience Scale (CD-RISC2), and Chronic Pain Acceptance Questionnaire (CPAQ-8) and telescoping, measured by a newly developed app. The participant completed a semi-structured interview that was designed to ascertain patterns in the overlapping experience of phantom limb telescoping and post-amputation pain.

Results

The participant rated his average PLP as 10 on a Numeric Rating Scale (NRS) from 0 (“no pain”) to 10 (“worst pain imaginable”) shortly after amputation. Approximately 12 months later, the participant noticed a shortening of his phantom limb, with a concurrent decrease in PLP. At present, his average NRS pain intensity is a 5/10. The participant described how the daily, debilitating PLP intensity diminished to weekly, manageable pain over time. Most notably, his responses on questionnaires were consistent with neuropathic PLP, mild to moderate levels of pain interference, a high level of catastrophic thinking about pain, low optimism, and mild symptoms of anxiety and depression.

Discussion

In this report, telescoping appeared to be preceded by an initial reduction in PLP intensity but these findings are based on a single case report and must be replicated with a large sample size before we have a clearer idea of the relationship between telescoping and PLP. This study provides insight into factors that may maintain PLP, generating targets for further investigation.
Isoliquiritigenin as a modulator of the Nrf2 signaling pathway: potential therapeutic implications
Authors: Qiu Mangmang , Ma Kang , Zhang Junfeng , Zhao Zhaohua , Wang Shan , Wang Qing , Xu Hao
Published: October 2024

Nuclear factor erythroid-2-related factor 2 (Nrf2), a transcription factor responsible for cytoprotection, plays a crucial role in regulating the expression of numerous antioxidant genes, thereby reducing reactive oxygen species (ROS) levels and safeguarding cells against oxidative stress. Extensive research has demonstrated the involvement of Nrf2 in various diseases, prompting the exploration of Nrf2 activation as a potential therapeutic approach for a variety of diseases. Consequently, there has been a surge of interest in investigating the Nrf2 signaling pathway and developing compounds that can modulate its activity. Isoliquiritigenin (ISL) (PubChem CID:638278) exhibits a diverse range of pharmacological activities, including antioxidant, anticancer, and anti-tumor properties. Notably, its robust antioxidant activity has garnered significant attention. Furthermore, ISL has been found to possess therapeutic effects on various diseases, such as diabetes, cardiovascular diseases, kidney diseases, and cancer, through the activation of the Nrf2 pathway. This review aims to evaluate the potential of ISL in modulating the Nrf2 signaling pathway and summarize the role of ISL in diverse diseases prevention and treatment through modulating the Nrf2 signaling pathway.
Identification of high-affinity Monoamine oxidase B inhibitors for depression and Parkinson’s disease treatment: bioinformatic approach of drug repurposing
Authors: Shahwan Moyad , Prasad Pratibha , Yadav Dharmendra Kumar , Altwaijry Nojood , Khan Mohd Shahnawaz , Shamsi Anas
Published: October 2024

Depression and Parkinson’s disease (PD) are devastating psychiatric and neurological disorders that require the development of novel therapeutic interventions. Drug repurposing targeting predefined pharmacological targets is a widely use approach in modern drug discovery. Monoamine oxidase B (MAO-B) is a critical protein implicated in Depression and PD. In this study, we undertook a systematic exploration of repurposed drugs as potential inhibitors of MAO-B. Exploring a library of 3,648 commercially available drug molecules, we conducted virtual screening using a molecular docking approach to target the MAO-B binding pocket. Two promising drug molecules, Brexpiprazole and Trifluperidol, were identified based on their exceptional binding potential and drug profiling. Subsequently, all-atom molecular dynamics (MD) simulations were performed on the MAO-B-ligand complexes for a trajectory of 300 nanoseconds (ns). Simulation results demonstrated that the binding of Brexpiprazole and Trifluperidol induced only minor structural alterations in MAO-B and showed significant stabilization throughout the simulation trajectory. Overall, the finding suggests that Brexpiprazole and Trifluperidol exhibit strong potential as repurposed inhibitors of MAO-B that might be explored further in experimental investigations for the development of targeted therapies for depression and PD.
Pharmacogenetics testing for poor response to antidepressants: a transnosographic case series
Authors: Lorvellec Marie-Agnès , Sipahimalani Gilles , Lahutte Bertrand , Delacour Hervé , Baldacci Antoine , Saguin Emeric
Published: October 2024

Introduction

Pharmacogenetics (PGx) holds promise for optimizing psychotropic medication use, with CYP2D6 and CYP2C19 identified as key genes in antidepressant treatment. However, few studies have explored the genetic variants of these genes in real-world settings for patients experiencing ineffectiveness or adverse drug reactions (ADRs) to antidepressants.

Methods

This case series includes 40 patients who underwent PGx testing due to antidepressant ineffectiveness or ADRs between June 2020 and April 2022. We describe the patients' demographic, clinical, and genetic characteristics and assess the value of PGx testing based on feedback from their psychiatrists.

Results

The most common diagnoses were major depressive disorder (60.0%) and post-traumatic stress disorder (30.0%). Ineffectiveness was reported in 65.0% of patients, ADRs in 2.5%, and both in 32.5%. The antidepressants involved included SSRIs (45.0%), SNRIs (27.5%), atypical antidepressants (20.0%), and tricyclics (17.5%). Only 17.5% of patients had normal CYP2D6 and CYP2C19 metabolic activity. Actionable genetic variants were identified in 22.0% of CYP2D6/CYP2C19-antidepressant-response pairs. PGx recommendations were followed in 92.7% of cases, with significant improvement in ADRs reported in 71.4% of patients and efficacy improvement in 79.5%.

Discussion

Our findings suggest that PGx testing can guide prescribing decisions for patients with antidepressant ineffectiveness or ADRs. The relatively high prevalence of genetic variants affecting pharmacokinetics supports the broader adoption of PGx testing in psychiatric practice.
Sorbaria sorbifolia flavonoid derivative induces mitochondrial apoptosis in human hepatoma cells through Bclaf1
Authors: Chen Jiaxin , Cheng Haoyi , Bai Chunhua , Wang Dandan , Fu Jinghao , Hao Jinge , Wang Yixuan , Xuewu Zhang
Published: October 2024

4′,5,7-Trihydroxy-8-methoxyflavone is an anticancer monomer component isolated from the traditional Chinese medicine Sorbaria sorbifolia. 4′,5-Dihydroxy-7-piperazinemethoxy-8-methoxy flavonoids (DMF) with good solubility and anti-tumor effects was obtained by chemical modification in the early stage. This study explored the mechanism by which DMF regulates the mitochondrial apoptosis of human hepatoma cells through Bcl-2-associated transcription factor 1 (Bclaf1). DMF inhibited the proliferation of human hepatoma cells in a concentration- and time-dependent manner and induced cell mitochondrial apoptosis. The molecular docking and cell assay results demonstrated that DMF inhibits Bclaf1 expression by binding to its active site. Lentivirus transfection was used to construct cells with stable knockout and overexpression of Bclaf1, and a Hep3B xenograft model was constructed in nude mice. The mechanism by which DMF induced the mitochondrial apoptosis of human hepatoma cells through Bclaf1 was further verified in vitro and in vivo. These findings indicated that DMF induced human hepatoma cell mitochondrial apoptosis through Bclaf1.
Induction of cytochrome P450 via upregulation of CAR and PXR: a potential mechanism for altered florfenicol metabolism by macranthoidin B in vivo
Authors: Li Si-cong , Wang Bin , Zhang Min , Yin Qin , Yang Zi-yi , Li Xu-ting , Liang Ge
Published: October 2024

Introduction

Macranthoidin B (MB) is a primary active component of Flos Lonicerae. In Chinese veterinary clinics, Flos Lonicerae is frequently used in combination with florfenicol to prevent and treat infections in livestock and poultry. However, potential interactions between Flos Lonicerae and florfenicol remain unclear. To systematically study these interactions, it is crucial to investigate the individual phytochemicals within Flos Lonicerae. Therefore, MB was selected for this study to assess its effect on the pharmacokinetics of florfenicol in vivo and to explore the underlying mechanisms involved.

Methods

Male Sprague-Dawley rats were administered MB (60 mg/kg BW) or sterile water orally for 7 consecutive days. On the 8th day, a single oral dose of florfenicol (25 mg/kg BW) was given. Florfenicol pharmacokinetics were analyzed using ultra-high performance liquid chromatography. The hepatic expression levels of cytochrome P450 (CYP1A2, CYP2C11, CYP3A1), UDP-glucuronosyltransferase (UGT1A1), P-glycoprotein (P-gp), and nuclear receptors, including constitutive androstane receptor (CAR), pregnane X receptor (PXR), and retinoid X receptor alpha (RXRα), were quantified via reverse transcription-quantitative polymerase chain reaction and Western blotting (WB). Hepatic CYP1A2 and CYP2C11 activities were measured using a cocktail method. Additionally, the subcellular expression and localization of CAR, PXR, and RXRαin hepatocytes was assessed using WB and immunofluorescence staining.

Results

MB significantly reduces the AUC(0-∞) and MRT(0-∞) of florfenicol. MB also markedly upregulates the mRNA and protein expression of hepatic CYP1A2 and CYP2C11, along with their catalytic activities. Substantial upregulation of CAR and PXR proteins occurs in the hepatocyte nucleus, along with significant nuclear colocalization of the transcriptionally active CAR/RXRα and PXR/RXRαheterodimers, indicating MB-induced nuclear translocation of both CAR and PXR.

Discussion

These findings suggest that MB-induced alterations in florfenicol pharmacokinetics, particularly its accelerated elimination, may be due to increased expression and activities of CYP1A2 and CYP2C11, with CAR and PXR potentially involved in these regulatory effects. Further investigation is yet needed to fully elucidate the clinical implications of these interactions concerning the efficacy of florfenicol in veterinary medicine.
Formulation development, characterization, and evaluation of sorafenib-loaded PLGA–chitosan nanoparticles
Authors: Mateen Abdul , Khan Abad , Khan Ismail , Ahmad Lateef , Khan Amjad , Salam Abdul
Published: October 2024

The basic purpose of this work was to develop environmentally friendly, biodegradable, and biocompatible polymeric nanoparticles of sorafenib that can effectively release the desired drug in a customized and controlled manner for targeting hepatocellular carcinoma. The solvent evaporation technique was employed for the synthesis of sorafenib-loaded PLGA–chitosan nanoparticles, followed by various experimental specifications and compatibility studies using poloxamer 407 as the stabilizer. The best nanoparticles thus synthesized were selected to be used for cytotoxicity investigations through in vitro and in vivo assessments. For the in vitro drug release tests, the dialysis bag diffusion technique was used. For both chitosan nanoparticles and PLGA loaded with sorafenib, a biphasic release pattern was found, exhibiting a protracted release lasting 10 days after a 24-h burst release. As experimental animals, rabbits were utilized to evaluate different in vivo pharmacokinetic properties of the selected formulations. Plasma samples were extracted with acetonitrile and analyzed through the developed HPLC method. Pharmacokinetic parameters such as AUC0-t, Cmax MRT, Vd, and half-life (t1/2) were enhanced significantly (p ≤ 0.001), while clearance was considerably decreased (p ≤ 0.001) for the chosen synthesized nanoparticles in contrast to the commercially accessible sorafenib formulation (Nexavar®). The cytotoxicity of the reference drug and sorafenib-loaded PLGA and chitosan nanoparticles was calculated by performing an MTT assay against HepG2 cell lines. The developed polymeric sorafenib nanoformulations possess the appropriate physicochemical properties, better targeting, surface morphology, and prolonged release kinetics. The pharmacokinetic parameters were improved significantly when the results were compared with commercially available sorafenib formulations.
The diversity of AMPA receptor inhibition mechanisms among amidine-containing compounds
Authors: Zhigulin Arseniy S. , Dron Mikhail Y. , Barygin Oleg I. , Tikhonov Denis B.
Published: October 2024

Amidine-containing compounds are primarily known as antiprotozoal agents (pentamidine, diminazene, furamidine) or as serine protease inhibitors (nafamostat, sepimostat, camostat, gabexate). DAPI is widely recognized as a fluorescent DNA stain. Recently, it has been shown that these compounds also act as NMDA receptor inhibitors. In this study, we examined the activity of these compounds and analyzed the mechanisms of action in relation to another important class of ionotropic glutamate receptors–calcium-permeable AMPA receptors (CP-AMPARs) and calcium-impermeable AMPA receptors (CI-AMPARs) – using the whole-cell patch-clamp method on isolated male Wistar rat brain neurons. Gabexate and camostat were found to be inactive. Other compounds preferentially inhibited calcium-permeable AMPA receptors with IC50 values of 30–60 µM. DAPI and furamidine were also active against CI-AMPARs with IC50s of 50–60 μM, while others showed poor activity. All active compounds acted as channel blockers, which are able for permeating into the cytoplasm on both CP- and CI-AMPARs. Specifically, sepimostat showed trapping in the closed CP-AMPAR channel. Furamidine and DAPI demonstrated a voltage-independent action on CI-AMPARs, indicating binding to an additional superficial site. While the majority of compounds inhibited glutamate-activated steady-state currents as well as kainate-activated currents on CI-AMPARs, pentamidine significantly potentiated glutamate-induced steady-state responses. The potentiating effect of pentamidine resembles the action of the positive allosteric modulator cyclothiazide although the exact binding site remains unclear. Thus, this study, together with our previous research on NMDA receptors, provides a comprehensive overview of this novel group of ionotropic glutamate receptors inhibitors with a complex pharmacological profile, remarkable diversity of effects and mechanisms of action.
Fentanyl enhances immune cell response through TLR4/MD-2 complex
Authors: Chemello Chiara , Facci Laura , Marcolin Emma , Ramaschi Giovanni Eugenio , Barbierato Massimo , Giusti Pietro , Bolego Chiara , Zusso Morena
Published: October 2024

Introduction

Opioids have been shown to induce neuroinflammation and immune cell activation, that might contribute to some of the opioid side effects, such as opioid-induced tolerance and paradoxical hyperalgesia. In this context, TLR4/MD-2 complex has been proposed as an off-target site for opioid action. This study was aimed at investigating the effect of fentanyl on lipopolysaccharide (LPS)-induced TLR4/MD-2 activation in rat primary microglia and human monocyte-derived macrophages (MDM).

Materials and Methods

The effect of fentanyl was first explored by measuring the expression and release of different proinflammatory mediators in primary rat microglia and human MDM by real-time PCR and ELISA. Then, the involvement of TLR4/MD-2 signaling was investigated studying NF-κB activation in HEK293 cells stably transfected with human TLR4, MD-2, and CD14 genes (HEK-Blue hTLR4 cells) and in human MDM.

Results

Fentanyl increased mRNA levels, as well as the LPS-induced secretion of proinflammatory mediators in primary microglia and MDM. Two inhibitors of TLR4/MD-2 signaling, namely the oxazoline derivative of N-palmitoylethanolamine (PEA-OXA) and CLI-095, blocked the production and release of proinflammatory cytokines by microglia stimulated with LPS and fentanyl, suggesting that TLR4/MD-2 could be the target of the proinflammatory activity of fentanyl. Finally, we showed that fentanyl in combination with LPS activated NF-κB signaling in human MDM and in HEK-Blue hTLR4 cells and this effect was blocked by inhibitors of TLR4/MD-2 complex.

Discussion

These results provide new insight into the mechanism of the proinflammatory activity of fentanyl, which involves the activation of TLR4/MD-2 signaling. Our findings might facilitate the development of novel inhibitors of TLR4/MD-2 signaling to combine with opioid-based analgesics for effective and safe pain management.
Exaggerated postnatal surge of orexin neurons and the effects of elimination of excess orexin on blood pressure and exaggerated chemoreflex in spontaneously hypertensive rats
Authors: Lusk Savannah , Moushey Alexander M. , Iwakoshi Nicholas , Wilson Christopher G. , Li Aihua , Ray Russell
Published: October 2024

An overactive orexin (OX) system is associated with neurogenic hypertension and an exaggerated chemoreflex in spontaneously hypertensive rats (SHRs). However, the chronology and mechanism of this association is unclear. We hypothesized that increased postnatal neurogenesis of OX neurons in SHRs precedes and contributes to the aberrant increase in mean arterial blood pressure (MAP) and the exaggerated response to hypercapnia during postnatal development. Using immunohistochemical methods and bromodeoxyuridine, we mapped the timeline of orexin neuron neurogenesis and maturation during early postnatal development. We then used whole-body plethysmography with EEG and EMG to map the development of mean arterial pressure (MAP) and state regulation. Finally, we used OX-targeted saporin toxin to determine the effects of eliminating excess OX neurons on the elevated MAP and exaggerated chemoreflex in adult SHRs. We found that both SHRs and Wistar–Kyoto (WKY) rats experienced postnatal increases in OX neurons. However, SHRs experienced a greater increase than WKY rats before P15, which led to significantly more OX neurons in SHRs than age-matched WKY controls by P15–16 (3,720 ± 780 vs. 2,406 ± 363, p = 0.005). We found that neurogenesis, as evidenced by BrdU staining in OX-positive neurons, was the primary contributor to the excess OX neurons in SHRs during early postnatal development. While SHRs develop more OX neurons by P15–16, SHRs and normotensive WKY control rats have similar MAP during postnatal development until P25 in wakefulness (81.6 ± 6.6 vs. 67.5 ± 6.8 mmHg, p = 0.006) and sleep (79.3 ± 6.1 vs. 66.6 ± 6.5, p = 0.009), about 10 days after the surge of OX neurons. By selectively eliminating excess (∼30%) OX neurons in SHRs, we saw a significantly lowered MAP and hypercapnic ventilatory chemoreflex compared to non–lesioned SHRs at P40. Additionally, we found unique signatures in state indicative of the attention defecit phenotype commonly associated with this model. We suggest that the postnatal increase of OX neurons, primarily attributed to exaggerated postnatal OX neurogenesis, may be necessary for the development of higher MAP and exaggerated chemoreflex in SHRs, and modulation of the overactive OX system may have a potential therapeutic effect during the pre-hypertensive period.
Comparative metabolomic analysis of spaghetti meat and wooden breast in broiler chickens: unveiling similarities and dissimilarities
Authors: Choi Janghan , Shakeri Majid , Kim Woo Kyun , Kong Byungwhi , Bowker Brian , Zhuang Hong
Published: October 2024

Introduction

Spaghetti meat (SM) and wooden breast (WB) are emerging myopathies in the breast meat of fast-growing broiler chickens. The purpose of the study was to investigate the metabolomic differences between normal (N), SM, and WB fillets 24 h postmortem.

Materials and methods

Eight chicken breasts for each experimental group were collected from a commercial processing plant. Supernatant from tissue homogenates were subjected to ultra-performance liquid chromatographytandem mass spectrometry (UPLC-MS) analysis.

Results and methods

A total of 3,090 metabolites were identified in the chicken breast meat. The comparison of WB and N showed 850 differential metabolites (P < 0.05), and the comparison of SM and N displayed 617 differential metabolites. The comparison of WB and SM showed 568 differential metabolites. The principal component analysis (PCA) plots showed a distinct separation between SM and N and between WB and N except for one sample, but SM and WB were not distinctly separated. Compared to N, 15-Hydroxyeicosatetraenoic acid (15-HETE) increased, and D-inositol-4-phosphate decreased in both SM and WB, indicating that cellular homeostasis and lipid metabolism can be affected in SM and WB. The abundance of nicotinamide adenine dinucleotide (NAD) + hydrogen (H) (NADH) was exclusively decreased between SM and N (P < 0.05). Purine metabolism was upregulated in SM and WB compared to N with a greater degree of upregulation in WB than SM. Folic acid levels decreased in SM and WB compared to N (P < 0.05). Steroid hormone biosynthesis was downregulated in SM compared to N (P < 0.05). Carbon metabolism was downregulated in SM and WB compared to N with greater degree of downregulation in WB than SM (P < 0.05). These data suggest both shared and unique metabolic alterations in SM and WB, indicating commonalities and differences in their underlying etiologies and meat quality traits. Dietary supplementation of deficient nutrients, such as NADH, folic acids, etc. and modulation of altered pathways in SM and WB would be strategies to reduce the incidence and severity of SM and WB.
Identification of genetic loci for powdery mildew resistance in common wheat
Authors: Liu Xia , Zhang Xiaoqing , Meng Xianghai , Liu Peng , Lei Menglin , Jin Hui , Wang Yanzhen , Jin Yirong , Cui Guoqing , Mu Zhixin , Liu Jindong , Jia Xiaoyun
Published: October 2024

Powdery mildew (PM) poses an extreme threat to wheat yields and quality. In this study, 262 recombinant inbred lines (RILs) of Doumai and Shi 4185 cross were used to map PM resistance genes across four environments. High-density genetic linkage map of the Doumai/Shi 4185 RIL population was constructed using the wheat Illumina iSelect 90K single-nucleotide polymorphism (SNP) array. In total, four stable quantitative trait loci (QTLs) for PM resistance, QPm.caas-2AS, QPm.caas-4AS, QPm.caas-4BL, and QPm.caas-6BS, were detected and explained 5.6%–15.6% of the phenotypic variances. Doumai contributed all the resistance alleles of QPm.caas-2AS, QPm.caas-4AS, QPm.caas-4BL, and QPm.caas-6BS. Among these, QPm.caas-4AS and QPm.caas-6BS overlapped with the previously reported loci, whereas QPm.caas-2AS and QPm.caas-4BL are potentially novel. In addition, six high-confidence genes encoding the NBS-LRR-like resistance protein, disease resistance protein family, and calcium/calmodulin-dependent serine/threonine-kinase were selected as the candidate genes for PM resistance. Three kompetitive allele-specific PCR (KASP) markers, Kasp_PMR_2AS for QPm.caas-2AS, Kasp_PMR_4BL for QPm.caas-4BL, and Kasp_PMR_6BS for QPm.caas-6BS, were developed, and their genetic effects were validated in a natural population including 100 cultivars. These findings will offer valuable QTLs and available KASP markers to enhance wheat marker-assisted breeding for PM resistance.
Physiological and metabolomic analyses reveal the mechanism by which exogenous spermine improves drought resistance in alfalfa leaves (Medicago sativa L.)
Authors: Wang Wenjuan , Kang Wenjuan , Shi Shangli , Liu Linbo
Published: October 2024

Introduction

Alfalfa (Medicago sativa L.) is a globally important legume crop with high nutritional and ecological value. Drought poses a serious threat to alfalfa acreage and yields. Spermine (Spm) has been shown to protect plants from drought damage. The aim of this study was to clarify the mechanism of exogenous Spm to improve drought resistance of alfalfa.

Methods

In this study, we root applied 0.1, 0.5, and 1 mM Spm to Gannong No. 3 (G3) alfalfa under drought stress, and then determined their physiological and metabolic changes.

Results

The results showed that exogenous Spm increased chlorophyll content, chlorophyll fluorescence parameters and gas exchange parameters, enhanced antioxidant enzymes activity, improved ascorbic acid-glutathione (AsA-GSH) cycle, increased osmoregulatory substances content, reduced hydrogen peroxide and superoxide anion levels, and inhibited malondialdehyde accumulation in alfalfa under drought stress, thereby increasing plant height and leaf relative water content and enhancing drought tolerance of alfalfa. The redundancy analysis of the above physiological indicators showed that the addition of the optimal Spm to improve drought tolerance of alfalfa under drought stress was mainly achieved by increasing catalase activity and improving the ASA-GSH cycle. In addition, metabolomics analysis revealed that exogenous Spm increased the content of oxobutanedioic acid, citric acid, fumaric acid and malic acid to enhance the tricarboxylic acid cycle. Meanwhile, exogenous Spm increased endogenous Spm and proline (Pro) content to resist drought stress by enhancing Spm and Pro metabolism. Moreover, exogenous Spm increased the accumulation of the signaling substance abscisic acid.

Discussion

In conclusion, exogenous Spm enhanced drought resistance of alfalfa leaves under drought stress.
Radical climate movements—is the hype about “eco-terrorism” analogy, warning or propaganda?
Authors: Lederer Markus , Lasso Mena Verena , Marquardt Jens , Richter Timo Alexander , Schoppek Dorothea Elena
Published: October 2024

Is the hype about “ecoterrorism” analogy, warning or propaganda? In order to answer this question, we start by defining radicalization, terrorism, and civil disobedience to develop systematic categories which allow us to pursue two specific research goals: First, we analyse how the breadth of the German climate movement is represented in the media, how the issue of “terrorism” is taken up and with what consequences for the debate. Here we make a discursive argument. Secondly, we use the information provided by the media reports, triangulate it with primary data from the movements analysed and secondary data from academic publications in order to assess the validity of the accusation of terrorism. Here we make a factual argument about the current properties of the climate movement. Finally, we bring both arguments together and argue that even the more radical currents of climate activism should not be classified as terrorists. What we can see is that there has been an attempt to criminalize demands of the radical climate movement during which large parts of the German print media have become willing handmaidens in the delegitimization of more or less radical climate groups. More recently, very first signs of a backlash against the criminalization can be detected.
Testing, revision and application of the social anxiety scale for Chinese social media users
Authors: Wang Yaning , Zhang Huan , Bao Sarenna
Published: October 2024

This study aims to modify the Social Anxiety Scale for Social Media Users (SAS-SMU) to make it more suitable for assessing social anxiety among Chinese social media users, taking into account the unique cultural nuances and social media usage patterns in China. Consequently, a systematic approach was adopted, involving three distinct studies. Study 1 involved translating the English scale into Chinese, conducting interviews with Chinese social media users, and subsequently revising the scale items based on the interview data to ensure cultural appropriateness. Study 2 employed an online survey to collect data and examine the reliability and construct validity of the revised scale, including a two-phase approach: an exploratory factor analysis (EFA) was conducted with 500 participants to identify the underlying factor structure, and a subsequent confirmatory factor analysis (CFA) with 408 participants was used to cross-validate the results. Thus far, this study has developed a social anxiety scale for Chinese mobile social media users (SAS-CMSMU) consisted of 11 items on three factors: Interaction Anxiety, Privacy Concern Anxiety, and Shared Content Anxiety. Study 3 (N = 1,006) applied the SAS-CMSMU to assess social anxiety among Wechat users. The results indicated that participants showed a high level of overall social anxiety (M = 3.646 > 3). Specifically, Privacy Anxiety was the most pronounced, followed by Shared Content Anxiety and Interaction Anxiety. Moreover, significant statistical differences in social anxiety levels were found among WeChat users across gender, age, education, income, and relationship status.
Mixed methods research on reflective writing in teacher education
Authors: Gläser-Zikuda Michaela , Zhang Chengming , Hofmann Florian , Plößl Lea , Pösse Lisa , Artmann Michaela
Published: October 2024

In this paper, mixed methods research is presented and discussed in the context of research on reflective writing for the professional development of pre-service teachers. First, we present prominent theoretical frameworks to analyze reflective writings. Second, we review relevant methodological approaches of research on reflective writings in teacher education, such as qualitative, quantitative and mixed methods. Third, we present a study from our research lab combining qualitative content analysis and linguistic analyses as an example for a concurrent mixed method approach. The results of the qualitative content analysis indicated that 198 reflective writings of pre-service teachers were primarily descriptive and on a low level. Computational linguistic analyses revealed that affective and cognitive terminology utilization in reflective writing differed significantly across the different levels of reflection, with a higher frequency of such terms correlating with deeper levels of reflection. Thus, essential challenges and opportunities of implementing such a mixed method study to analyze reflective writings are illustrated and discussed. Finally, we conclude the paper by discussing on how mixed methods approaches might be further advanced in the field of reflective writing research in teacher education.