bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Encapsulated Purple Garlic Oil on Microvascular Function and the Components of Metabolic Syndrome: A Randomized Placebo-Controlled Study—The ENDOTALLIUM Study
Authors: Bara-Ledesma Nuria , Jimenez-Esteban Judith , Fabregate Martin , Fabregate-Fuente Rosa , Cymberknop Leandro Javier , Castillo-Martinez Purificacion , Navarro-Fayos Maria Teresa , Gomez del Olmo Vicente , Saban-Ruiz Jose
Published: June 2024

Endothelial dysfunction (ED) is associated with progressive changes contributing to clinical complications related to macro- and microvascular diseases. Garlic (Allium sativum L.) and its organosulfur components have been related to beneficial cardiovascular effects and could improve endothelial function. The ENDOTALLIUM Study aimed to evaluate the effect of the regular consumption of encapsulated purple garlic oil on microvascular function, endothelial-related biomarkers, and the components of metabolic syndrome (MetS) in untreated subjects with cardiometabolic alterations. Fifty-two individuals with at least one MetS component were randomized (1:1) in a single-center, single-blind, placebo-controlled, parallel-group study. The participants received encapsulated purple garlic oil (n = 27) or placebo (n = 25) for five weeks. Skin microvascular peak flow during post-occlusive reactive hyperemia significantly increased in the purple garlic oil group compared to the placebo group (between-group difference [95%CI]: 15.4 [1.5 to 29.4] PU; p = 0.031). Likewise, hs-CRP levels decreased in the purple garlic group compared to the control group (−1.3 [−2.5 to −0.0] mg/L; p = 0.049). Furthermore, we observed a significant reduction in the mean number of MetS components in the purple garlic group after five weeks (1.7 ± 0.9 vs. 1.3 ± 1.1, p = 0.021). In summary, regular consumption of encapsulated purple garlic oil significantly improved microvascular function, subclinical inflammatory status, and the overall MetS profile in a population with cardiometabolic alterations.
Pain Assessment in Patients during Hemodialysis Treatment: Quality Improvement Project
Authors: Rodrigues Rita , Costeira Cristina
Published: June 2024

Pain is a prevalent symptom in patients with chronic kidney disease, related to disease progression, comorbidities, and required immobility during dialysis treatment. Nurses must perform detailed pain assessments to manage pain effectively during hemodialysis treatment. This quality improvement project, reported using SQUIRE 2.0, aims to describe pain characteristics in chronic kidney disease patients in a hemodialysis unit clinic in Portugal, implement strategies to improve the pain assessment process in patients with chronic kidney disease during hemodialysis treatment, and assess nurses’ satisfaction with the implemented strategies. The study was conducted in a Portuguese hemodialysis clinic, with patients and nurses, in three phases: diagnostic study, protocol implementation, and a descriptive study to assess nurse satisfaction. Seventy-five patients (mean age 71 ± 12.6 years) participated, with 64% reporting moderate chronic pain daily and 48% during hemodialysis treatment. Thirteen nurses considered the pain assessment protocol important, rating their satisfaction at 7.92 ± 1.32 (0–10). Standardizing practices through protocol implementation is likely to improve care and increase nurse satisfaction.
Rapid Immune Modulation after Consuming Euglena gracilis Whole Algae Involving Altered Responses to Ex Vivo Immune Challenges: A Placebo-Controlled Cross-Over Trial
Authors: Iloba Ifeanyi , Cruickshank Dina , Sanchez Krista , Brawer Solli , Grundman Omer , Jensen Gitte S.
Published: June 2024

Euglena gracilis (EG) microalgae has immune-modulating properties, partly due to its unique intracellular β-glucan-granules (paramylon). We evaluated the effects of EG consumption on immune status in vivo, ex vivo, and in vitro. A placebo-controlled cross-over study evaluated acute immune surveillance, followed by a 1-week open-label phase. Immune training was documented using ex vivo immune challenges and cytokine profiles. In vitro testing of monocytes compared the effects of EG to pure β-glucan. Compared to placebo, EG consumption triggered increased T cell numbers in the blood circulation (1 h: p < 0.01) and decreased monocyte numbers (2 h: p < 0.05). Natural killer cells showed increased CD25 expression (1 and 2 h: p < 0.01) and reduced CD69 expression (2 h: p < 0.01). T cells showed reduced CD25 and CD69 expression (p < 0.01). There were no significant changes to serum cytokines. After EG consumption, ex vivo cultures of peripheral blood mononuclear cells showed significant changes to spontaneous and inflammation-induced cytokine levels after 2 h (increased G-CSF: p < 0.01, reduced IL-1β and TNF-α (p < 0.05)) and one week (reduced TNF-α (p < 0.01) and increased IL-10 (p < 0.05)). In vitro, EG-trained monocytes responded differently to a second stimulus than β-glucan-trained monocytes (increased IL-1b: p < 0.1, TNF-α: p < 0.01). EG-mediated training of innate immunity, combined with long-term modulation of inflammation, suggests a nutraceutical strategy for preventive immune support.
Investigation of SARS-CoV-2 Infection among Companion Animals in Households with Confirmed Human COVID-19 Cases
Authors: Venkat Heather , Yaglom Hayley D. , Hecht Gavriella , Goedderz Andrew , Ely Jennifer L. , Sprenkle Michael , Martins Taylor , Jasso-Selles Daniel , Lemmer Darrin , Gesimondo Jordan , Ruberto Irene , Komatsu Kenneth , Engelthaler David M.
Published: June 2024

We aimed to characterize SARS-CoV-2 infection in companion animals living in households with COVID-19-positive people and understand the dynamics surrounding how these animals become infected. Public health investigators contacted households with at least one confirmed, symptomatic person with COVID-19 for study recruitment. Blood, nasal, and rectal swab specimens were collected from pet dogs and cats and a questionnaire was completed. Specimens were tested for SARS-CoV-2 by RT-PCR, and for neutralizing antibodies; genomic sequencing was performed on viral-positive samples. A total of 36.4% of 110 pets enrolled had evidence of infection with SARS-CoV-2. Pets were more likely to test positive if the pet was immunocompromised, and if more than one person in the home was positive for COVID-19. Among 12 multi-pet households where at least one pet was positive, 10 had at least one other pet test positive. Whole-genome sequencing revealed the genomes of viral lineages circulating in the community during the time of sample collection. Our findings suggest a high likelihood of viral transmission in households with multiple pets and when pets had very close interactions with symptomatic humans. Further surveillance studies are needed to characterize how new variants impact animals and to understand opportunities for infection and spillover in susceptible species.
Sex Differences during Influenza A Virus Infection and Vaccination and Comparison of Cytokine and Antibody Responses between Plasma and Serum Samples
Authors: Dhakal Santosh , Wolfe Brian W. , Pantha Saurav , Vijayakumar Saranya
Published: June 2024

In this study, we evaluated sex differences during infection with mouse-adapted H1N1 and H3N2 influenza A viruses (IAVs) in the C57BL/6J mouse model and compared the cytokine and antibody responses between plasma and serum samples during IAV infection and vaccination. Lethal doses for both H1N1 and H3N2 IAVs were lower for adult females and they suffered with greater morbidity than adult males when infected with sublethal doses. In influenza virus-infected mice, cytokine responses differed between plasma and serum samples. After inactivated influenza virus vaccination and drift variant challenge, adult female mice had greater antibody responses and were better protected. In influenza-vaccinated and challenged mice, binding antibodies were unaffected between paired plasma or serum samples. However, functional antibody assays, including hemagglutination inhibition, microneutralization, and antibody-dependent cellular cytotoxicity assays, were affected by the use of plasma and serum sample types. Our results indicate that careful consideration is required while selecting plasma versus serum samples to measure cytokine and antibody responses during IAV infection and vaccination.
Oropouche Virus Exposure in Febrile Patients during Chikungunya Virus Introduction in the State of Amapá, Amazon Region, Brazil
Authors: de Lima Raquel Curtinhas , Dias Helver Gonçalves , de Souza Thiara Manuele Alves , Familiar-Macedo Débora , Ribeiro Edcelha D’Athaide , Corrêa Valmir Corrêa e , Pauvolid-Corrêa Alex , de Azeredo Elzinandes Leal , dos Santos Flávia Barreto
Published: June 2024

Oropouche orthobunyavirus (OROV) is an arbovirus transmitted by midges that has been involved in outbreaks throughout Central and South America. In Brazil, human cases have been historically concentrated in the northern region of the country. Oropouche fever in humans range from mild clinical signs to rare neurological events, and is considered a neglected tropical disease in Brazil. Due to the clinical similarities to other arboviruses, such as chikungunya and dengue viruses, OROV infections are likely to be underreported. Chikungunya virus (CHIKV) cases in Brazil were first recognized in 2014 in the states of Amapá and Bahia in the north and northeast regions, respectively. Both OROV and CHIKV cause nonspecific symptoms, making clinical diagnosis difficult in a scenario of arbovirus cocirculation. Aiming to investigate OROV transmission during the CHIKV introduction in the state of Amapá located in the Brazilian Amazon, we conducted a retrospective molecular (RT-qPCR) and serological investigation in febrile cases (N = 166) collected between August 2014 and May 2015. All acute serum samples were negative for OROV RNA using RT-qPCR. However, neutralizing antibodies for OROV were detected using a plaque reduction neutralization test (PRNT90) in 10.24% (17/166) of the patients, with neutralizing antibody titers ranging from 20 to ≥640, suggesting the previous exposure of patients to OROV. Regarding CHIKV, recent exposure was confirmed by the detection of CHIKV RNA in 20.25% (33/163) of the patients and by the detection of anti-CHIKV IgM in 28.57% (44/154) of the patients. The additional detection of anti-CHIKV IgG in 12.58% (19/151) of the febrile patients suggests that some individuals had been previously exposed to CHIKV. Whether the OROV exposure reported here occurred prior or during the CHIKV circulation in Amapá, is unknown, but because those arboviral infections share similar clinical signs and symptoms, a silent circulation of enzootic arboviruses during the introduction of exotic arboviruses may occur, and highlights the importance of syndromic cases’ surveillance to arboviruses in Brazil.
Chromosomal Integration of HHV-6 in a Preterm Neonate: A Rare Case of Hyperleukocytosis and Clinical Implications
Authors: Balasundaram Palanikumar , Sakr Mohamed
Published: June 2024

Leukocytosis in neonates can occur because of infectious, inflammatory, malignant, or physiological processes. Hyperleukocytosis is defined as a total leukocyte count (TLC) exceeding 100,000 per mm3, warranting immediate evaluation. Neonates with hyperleukocytosis are at risk of leukostasis and the associated severe complications, including respiratory distress, myocardial ischemia, hyperuricemia, acute renal failure, infarction, and hemorrhage. Differentiating leukemia and leukemoid reactions in neonates presenting with elevated TLC is challenging but critical. We present a unique case of a preterm male neonate with hyperleukocytosis, initially suspected to have an underlying malignancy. The neonate’s clinical course was complicated by respiratory distress syndrome and anemia of prematurity, necessitating neonatal intensive care unit management. Further investigation revealed high human herpesvirus 6 (HHV-6) DNA levels in the whole blood, leading to a chromosomally integrated HHV-6 (ciHHV-6) diagnosis. CiHHV-6 is characterized by HHV-6 DNA integration into the host genome. Accurate diagnosis relies on whole-blood quantitative PCR, distinguishing ciHHV-6 from an active infection. The neonate remained asymptomatic, and antiviral treatment was deemed unnecessary. This case underscores the importance of recognizing ciHHV-6 as a potential cause of hyperleukocytosis in neonates and highlights the value of whole-blood PCR for differentiation. Understanding the spectrum of HHV-6 infection in neonates is vital for appropriate management and prognostication.
Meta-Analysis of the Safety and Efficacy of Direct Oral Anticoagulants for the Treatment of Left Ventricular Thrombus
Authors: Vorla Mounica , Kalra Dinesh K.
Published: June 2024

Background: Literature on the preferred anticoagulant for treating left ventricular thrombus (LVT) is lacking. Thus, our objective was to compare the efficacy of DOACs versus warfarin in treating LVT. Methods: Databases were searched for RCTs and adjusted observational studies that compared DOAC versus warfarin through March 2024. The primary efficacy outcomes of interest were LVT resolution, systemic embolism, composite of stroke, and TIA. The primary safety outcomes encompassed all-cause mortality and bleeding events. Results: Our meta-analysis including 31 studies demonstrated that DOAC use was associated with higher odds of thrombus resolution (OR: 1.08, 95% CI: 0.86–1.31, p: 0.46). A statistically significant reduction in the risk of stroke/TIA was observed in the DOAC group versus the warfarin group (OR: 0.65, 95% CI: 0.48–0.89, p: 0.007). Furthermore, statistically significant reduced risks of all-cause mortality (OR: 0.68, 95% CI: 0.47–0.98, p: 0.04) and bleeding events (OR: 0.70, 95% CI: 0.55–0.89, p: 0.004) were observed with DOAC use as compared to warfarin use. Conclusion: Compared to VKAs, DOACs are noninferior as the anticoagulant of choice for LVT treatment. However, further studies are warranted to confirm these findings.
Medication and Outcome in Older Heart Failure Patients: Results from a Prospective Cohort Study
Authors: Garay David Peter , Saner Hugo , Herzberg Jan , Hellige Gerrit , Arenja Nisha
Published: June 2024

Purpose: Acute heart failure (AHF) is associated with high morbidity and mortality, and the prognosis is particularly poor in older patients. Although the application of guideline-directed medical therapy (GDMT) has shown a positive impact on prognosis, the effects are less clear in older age groups. The aim of this study was to analyze real-world data regarding GDMT and outcomes in older HF patients. Methods: This is a prospective cohort study from a secondary care hospital in central Switzerland. A total of 97 consecutive patients aged ≥60 years were enrolled between January 2019 and 2022. The main outcome parameters were prescribed GDMT at discharge, and in case of rehospitalization, GDMT at readmission, and survival in terms of all-cause mortality and HF-related hospitalizations during a 3-year follow-up period. Results: Follow-up data were available for 93/97 patients. The mean age was 77.8 ± 9.8 years, 46% being female. The mean left ventricular ejection fraction (LVEF) was 35.3 ± 13.9%, with a mean BNP level of 2204.3 ± 239 ng/L. Upon discharge, 86% received beta-blockers and 76.3% received renin–angiotensin system (RAS) inhibitors. At rehospitalization for AHF, beta-blockers use was significantly lower and decreased to 52.8% (p = 0.003), whereas RAS inhibitor use increased slightly to 88.9% (p = 0.07), and SGLT-2 inhibitors showed a significant increase from 5.4% vs. 47.2% (p = 0.04). GDMT prescription was not dependent on LVEF. Overall, 73.1% of patients received two-stage or three-stage GDMT at discharge, whereas this percentage decreased to 61% at rehospitalization (p = 0.01). Kaplan–Meier analysis for the combined outcome rehospitalization and death stratified by LV function showed significant differences between LVEF groups (aHR: 0.6 [95% CI: 0.44 to 0.8]; p = 0.0023). Conclusions: Our results indicate that first, the majority of older AHF patients from a secondary care hospital in Switzerland were not on optimal GDMT at discharge and even fewer at readmission, and second, that prognosis of the population is still poor, with almost half of the patients having been rehospitalized or died during a 3-year follow-up period under real-world conditions, without significant difference between women and men. Our findings underline the need for further improvements in the medical treatment of AHF, in particular in older patients, to improve prognosis and to reduce the burden of disease.
The Role of Fibroblast Activation Protein Inhibitor Positron Emission Tomography in Inflammatory and Infectious Diseases: An Updated Systematic Review
Authors: Albano Domenico , Rizzo Alessio , Slart Riemer H. J. A. , Hess Søren , Noriega-Álvarez Edel , Wakfie-Corieh Cristina Gamila , Leccisotti Lucia , Glaudemans Andor W. J. M. , Gheysens Olivier , Treglia Giorgio
Published: June 2024

The role of fibroblast activation protein inhibitor (FAPI) positron emission tomography/computed tomography (PET/CT) is emerging for the assessment of non-oncological diseases, such as inflammatory and infectious diseases, even if the evidence in the literature is still in its initial phases. We conducted a systematic search of Scopus, PubMed/MEDLINE, Embase, and Cochrane library databases for studies published before 31 December 2023 reporting infectious and inflammatory disease imaging with FAPI PET/CT. We included twenty-one studies for a total of 1046 patients. The most frequent disease studied was lung interstitial disease, investigated in six studies for a total of 200 patients, followed by bone and joint diseases in two studies and 185 patients, IgG4-related disease in 53 patients, and Crohn’s disease in 30 patients. Despite the heterogeneity of studies in terms of study design and technical features, FAPI PET/CT showed a high detection rate and diagnostic role. Moreover, when compared with 2-[18F]FDG PET/CT (n = 7 studies), FAPI PET/CT seems to have better diagnostic performances. The presence of chronic inflammation and tissue remodeling, typical of immune-mediated inflammatory conditions, may be the underlying mechanism of FAPI uptake.
Potential of Bitter Medicinal Plants: A Review of Flavor Physiology
Authors: Zuluaga Germán
Published: June 2024

The function of the sense of taste is usually confined to the ability to perceive the flavor of food to assess and use the nutrients necessary for healthy survival and to discard those that may be harmful, toxic, or unpleasant. It is almost unanimously agreed that the perception of bitter taste prevents the consumption of toxins from plants, decaying foods, and drugs. Forty years ago, while practicing medicine in a rural area of the Colombian Amazon, I had an unexpected encounter with the Inga Indians. I faced the challenge of accepting that their traditional medicine was effective and that the medicinal plants they used had a real therapeutic effect. Wanting to follow a process of learning about medicinal plants on their terms, I found that, for them, the taste of plants is a primary and fundamental key to understanding their functioning. One of the most exciting results was discovering the therapeutic value of bitter plants. The present review aims to understand whether there is any scientific support for this hypothesis from the traditional world. Can the taste of plants explain their possible therapeutic benefit? In the last 20 years, we have made novel advances in the knowledge of the physiology of taste. Our purpose will be to explore these scientific advances to determine if the bitter taste of medicinal plants benefits human health.
Porous Chitosan/Hydroxyapatite Composite Microspheres for Vancomycin Loading and Releasing
Authors: Wu Meng-Ying , Kuo Yi-Ting , Kao I-Fang , Yen Shiow-Kang
Published: June 2024

Porous chitosan/hydroxyapatite (Chi-HAp) composite microspheres were prepared in an aqueous solution containing chitosan, calcium nitrate, and ammonium dihydrogen phosphate by using a hydrothermal method at various temperatures. The investigation indicated that temperature significantly impacted the final product’s appearance. Hydroxyapatite (HAp) coupled with dicalcium phosphate dihydrate (DCPD) flakes were obviously found at 65 and 70 °C, while the latter gradually disappeared at higher temperatures. Conversely, synthesis at 90 °C led to smaller particle sizes due to the broken chitosan chains. The microspheres synthesized at 75 °C were selected for further analysis, revealing porous structures with specific surface areas of 36.66 m2/g, pores ranging from 3 to 100 nm, and pore volumes of 0.58 cm3/g. Vancomycin (VCM), an antibiotic, was then absorbed on and released from the microspheres derived at 75 °C, with a drug entrapment efficiency of 20% and a release duration exceeding 20 days. The bacteriostatic activity of the VCM/composite microspheres against Staphylococcus aureus increased with the VCM concentration and immersion time, revealing a stable inhibition zone diameter of approximately 4.3 mm from 24 to 96 h, and this indicated the retained stability and efficacy of the VCM during the encapsulating process.
Study on the Effect of Pharmaceutical Excipient PEG400 on the Pharmacokinetics of Baicalin in Cells Based on MRP2, MRP3, and BCRP Efflux Transporters
Authors: Yang Dan , Zhang Min , Zhao Mei , Li Chaoji , Shang Leyuan , Zhang Shuo , Wang Pengjiao , Gao Xiuli
Published: June 2024

Pharmaceutical excipient PEG400 is a common component of traditional Chinese medicine compound preparations. Studies have demonstrated that pharmaceutical excipients can directly or indirectly influence the disposition process of active drugs in vivo, thereby affecting the bioavailability of drugs. In order to reveal the pharmacokinetic effect of PEG400 on baicalin in hepatocytes and its mechanism, the present study first started with the effect of PEG400 on the metabolic disposition of baicalin at the hepatocyte level, and then the effect of PEG400 on the protein expression of baicalin-related transporters (BCRP, MRP2, and MRP3) was investigated by using western blot; the effect of MDCKII-BCRP, MDCKII-BCRP, MRP2, and MRP3 was investigated by using MDCKII-BCRP, MDCKII-MRP2, and MDCKII-MRP3 cell monolayer models, and membrane vesicles overexpressing specific transporter proteins (BCRP, MRP2, and MRP3), combined with the exocytosis of transporter-specific inhibitors, were used to study the effects of PEG400 on the transporters in order to explore the possible mechanisms of its action. The results demonstrated that PEG400 significantly influenced the concentration of baicalin in hepatocytes, and the AUC0–t of baicalin increased from 75.96 ± 2.57 μg·h/mL to 106.94 ± 2.22 μg·h/mL, 111.97 ± 3.98 μg·h/mL, and 130.42 ± 5.26 μg·h/mL (p ˂ 0.05). Furthermore, the efflux rate of baicalin was significantly reduced in the vesicular transport assay and the MDCKII cell model transport assay, which indicated that PEG400 had a significant inhibitory effect on the corresponding transporters. In conclusion, PEG400 can improve the bioavailability of baicalin to some extent by affecting the efflux transporters and thus the metabolic disposition of baicalin in the liver.
Atomic Force Microscopy for the Study of Cell Mechanics in Pharmaceutics
Authors: Siboni Henrik , Ruseska Ivana , Zimmer Andreas
Published: June 2024

Cell mechanics is gaining attraction in drug screening, but the applicable methods have not yet become part of the standardized norm. This review presents the current state of the art for atomic force microscopy, which is the most widely available method. The field is first motivated as a new way of tracking pharmaceutical effects, followed by a basic introduction targeted at pharmacists on how to measure cellular stiffness. The review then moves on to the current state of the knowledge in terms of experimental results and supplementary methods such as fluorescence microscopy that can give relevant additional information. Finally, rheological approaches as well as the theoretical interpretations are presented before ending on additional methods and outlooks.
Comparative In Vitro Study: Assessing Phytochemical, Antioxidant, Antimicrobial, and Anticancer Properties of Vaccinium macrocarpon Aiton and Vaccinium oxycoccos L. Fruit Extracts
Authors: Šedbarė Rima , Janulis Valdimaras , Pavilonis Alvydas , Petrikaite Vilma
Published: June 2024

The phytochemical diversity and potential health benefits of V. oxycoccos and V. macrocarpon fruits call for further scientific inquiry. Our study aimed to determine the phytochemical composition of extracts from these fruits and assess their antioxidant, antibacterial, and anticancer properties in vitro. It was found that the ethanolic extracts of V. oxycoccos and V. macrocarpon fruits, which contained more lipophilic compounds, had 2–14 times lower antioxidant activity compared to the dry aqueous extracts of cranberry fruit, which contained more hydrophilic compounds. All tested cranberry fruit extracts (OE, OW, ME, and MW) significantly inhibited the growth of bacterial strains S. aureus, S. epidermidis, E. coli, and K. pneumoniae in vitro compared to the control. Cytotoxic activity against the human prostate carcinoma PPC-1 cell line, human renal carcinoma cell line (CaKi-1), and human foreskin fibroblasts (HF) was determined using an MTT assay. Furthermore, the effect of the cranberry fruit extract samples on cell migration activity, cancer spheroid growth, and viability was examined. The ethanolic extract from V. macrocarpon fruits (ME) showed higher selectivity in inhibiting the viability of prostate and renal cancer cell lines compared to fibroblasts. It also effectively hindered the migration of these cancer cell lines. Additionally, the V. macrocarpon fruit extract (ME) demonstrated potent cytotoxicity against PPC-1 and CaKi-1 spheroids, significantly reducing the size of PPC-1 spheroids compared to the control. These findings suggest that cranberry fruit extracts, particularly the ethanolic extract from V. macrocarpon fruits, have promising potential as natural remedies for bacterial infections and cancer therapy.
Applicability of MDR1 Overexpressing Abcb1KO-MDCKII Cell Lines for Investigating In Vitro Species Differences and Brain Penetration Prediction
Authors: Sóskuti Emőke , Szilvásy Nóra , Temesszentandrási-Ambrus Csilla , Urbán Zoltán , Csíkvári Olivér , Szabó Zoltán , Kecskeméti Gábor , Pusztai Éva , Gáborik Zsuzsanna
Published: June 2024

Implementing the 3R initiative to reduce animal experiments in brain penetration prediction for CNS-targeting drugs requires more predictive in vitro and in silico models. However, animal studies are still indispensable to obtaining brain concentration and determining the prediction performance of in vitro models. To reveal species differences and provide reliable data for IVIVE, in vitro models are required. Systems overexpressing MDR1 and BCRP are widely used to predict BBB penetration, highlighting the impact of the in vitro system on predictive performance. In this study, endogenous Abcb1 knock-out MDCKII cells overexpressing MDR1 of human, mouse, rat or cynomolgus monkey origin were used. Good correlations between ERs of 83 drugs determined in each cell line suggest limited species specificities. All cell lines differentiated CNS-penetrating compounds based on ERs with high efficiency and sensitivity. The correlation between in vivo and predicted Kp,uu,brain was the highest using total ER of human MDR1 and BCRP and optimized scaling factors. MDR1 interactors were tested on all MDR1 orthologs using digoxin and quinidine as substrates. We found several examples of inhibition dependent on either substrate or transporter abundance. In summary, this assay system has the potential for early-stage brain penetration screening. IC50 comparison between orthologs is complex; correlation with transporter abundance data is not necessarily proportional and requires the understanding of modes of transporter inhibition.
Applying Physiologically Based Pharmacokinetic Modeling to Interpret Carbamazepine’s Nonlinear Pharmacokinetics and Its Induction Potential on Cytochrome P450 3A4 and Cytochrome P450 2C9 Enzymes
Authors: Yin Xuefen , Cicali Brian , Rodriguez-Vera Leyanis , Lukacova Viera , Cristofoletti Rodrigo , Schmidt Stephan
Published: June 2024

Carbamazepine (CBZ) is commonly prescribed for epilepsy and frequently used in polypharmacy. However, concerns arise regarding its ability to induce the metabolism of other drugs, including itself, potentially leading to the undertreatment of co-administered drugs. Additionally, CBZ exhibits nonlinear pharmacokinetics (PK), but the root causes have not been fully studied. This study aims to investigate the mechanisms behind CBZ’s nonlinear PK and its induction potential on CYP3A4 and CYP2C9 enzymes. To achieve this, we developed and validated a physiologically based pharmacokinetic (PBPK) parent–metabolite model of CBZ and its active metabolite Carbamazepine-10,11-epoxide in GastroPlus®. The model was utilized for Drug–Drug Interaction (DDI) prediction with CYP3A4 and CYP2C9 victim drugs and to further explore the underlying mechanisms behind CBZ’s nonlinear PK. The model accurately recapitulated CBZ plasma PK. Good DDI performance was demonstrated by the prediction of CBZ DDIs with quinidine, dolutegravir, phenytoin, and tolbutamide; however, with midazolam, the predicted/observed DDI AUClast ratio was 0.49 (slightly outside of the two-fold range). CBZ’s nonlinear PK can be attributed to its nonlinear metabolism caused by autoinduction, as well as nonlinear absorption due to poor solubility. In further applications, the model can help understand DDI potential when CBZ serves as a CYP3A4 and CYP2C9 inducer.
Hybrid Nanosystem Formed by DOX-Loaded Liposomes and Extracellular Vesicles from MDA-MB-231 Is Effective against Breast Cancer Cells with Different Molecular Profiles
Authors: Paschoal Barbosa Luiza Marques , Gomes Eliza Rocha , Barros André Luis Branco de , Cassali Geovanni Dantas , Carvalho Andréa Teixeira de , Silva Juliana de Oliveira , Pádua Ana Luiza , Oliveira Mônica Cristina
Published: June 2024

Drug delivery selectivity is a challenge for cancer treatment. A hybrid pegylated pH-sensitive liposome–extracellular vesicle isolated from human breast cancer cell MDA-MB-231 was developed to investigate its in vitro activity against breast cancer cells of different molecular profiles to overcome this inconvenience. The hybrid nanosystem was produced by film hydration, and doxorubicin (DOX) was encapsulated in this system using the ammonium sulfate gradient method. The characterization of this hybrid nanosystem revealed a mean diameter of 140.20 ± 2.70 nm, a polydispersity index of 0.102 ± 0.033, an encapsulation efficiency of doxorubicin of 88.9% ± 2.4, and a great storage stability for 90 days at 4 °C. The fusion of extracellular vesicles with liposomes was confirmed by nanoflow cytometry using PE-conjugated human anti-CD63. This hybrid nanosystem demonstrated cytotoxicity against human breast cancer cell lines with different molecular subtypes, enhanced anti-migration properties, and exhibited similar cellular uptake to the free DOX treatment. Preliminary acute toxicity assessments using Balb/C female mice indicated a median lethal dose of 15–17.5 mg/kg, with no evidence of splenic, liver, heart, bone marrow, and renal damage at a dose of 15 mg/kg. These findings suggest the hybrid formulation as a versatile nanocarrier for the treatment of various breast cancer subtypes.
Modulation of Brain Cholesterol Metabolism through CYP46A1 Overexpression for Rett Syndrome
Authors: Audouard Emilie , Khefif Nicolas , Gillet-Legrand Béatrix , Nobilleau Fanny , Bouazizi Ouafa , Stanga Serena , Despres Gaëtan , Alves Sandro , Lamazière Antonin , Cartier Nathalie , Piguet Françoise
Published: June 2024

Rett syndrome (RTT) is a rare neurodevelopmental disorder caused by mutation in the X-linked gene methyl-CpG-binding protein 2 (Mecp2), a ubiquitously expressed transcriptional regulator. RTT results in mental retardation and developmental regression that affects approximately 1 in 10,000 females. Currently, there is no curative treatment for RTT. Thus, it is crucial to develop new therapeutic approaches for children suffering from RTT. Several studies suggested that RTT is linked with defects in cholesterol homeostasis, but for the first time, therapeutic evaluation is carried out by modulating this pathway. Moreover, AAV-based CYP46A1 overexpression, the enzyme involved in cholesterol pathway, has been demonstrated to be efficient in several neurodegenerative diseases. Based on these data, we strongly believe that CYP46A1 could be a relevant therapeutic target for RTT. Herein, we evaluated the effects of intravenous AAVPHP.eB-hCYP46A1-HA delivery in male and female Mecp2-deficient mice. The applied AAVPHP.eB-hCYP46A1 transduced essential neurons of the central nervous system (CNS). CYP46A1 overexpression alleviates behavioral alterations in both male and female Mecp2 knockout mice and extends the lifespan in Mecp2-deficient males. Several parameters related to cholesterol pathway are improved and correction of mitochondrial activity is demonstrated in treated mice, which highlighted the clear therapeutic benefit of CYP46A1 through the neuroprotection effect. IV delivery of AAVPHP.eB-CYP46A1 is perfectly well tolerated with no inflammation observed in the CNS of the treated mice. Altogether, our results strongly suggest that CYP46A1 is a relevant target and overexpression could alleviate the phenotype of Rett patients.
A Near Fourier-Limited Pulse-Preserving Monochromator for Extreme-Ultraviolet Pulses in the Few-Fs Regime
Authors: Yang Yudong , Neumann Tanja , Hengster Julia , Mainz Roland E. , Elsner Jakob , Mücke Oliver D. , Kärtner Franz X. , Uphues Thorsten
Published: June 2024

We present a pulse-preserving multilayer-based extreme-ultraviolet (XUV) monochromator providing ultra-narrow bandwidth (ΔE<0.6eV, Ec=92eV) and compact footprint (28×10cm2) for easy integration into high-harmonic generation (HHG) or free-electron laser (FEL) sources. The temporal resolution of the novel design supports pulse durations of typical pump–probe setups in the femtosecond and attosecond regime, depending on the mirror design and focusing geometries over the tuning range of the monochromator. The theoretical design is analyzed and experimentally characterized in a laser-driven HHG setup.
Far-Field Super-Resolution Microscopy Using Evanescent Illumination: A Review
Authors: Zhang Qianwei , Zhang Haonan , Yang Xiaoyu , Liu Xu , Tang Mingwei , Yang Qing
Published: June 2024

The resolution of conventional optical microscopy is restricted by the diffraction limit. Light waves containing higher-frequency information about the sample are bound to the sample surface and cannot be collected by far-field optical microscopy. To break the resolution limit, researchers have proposed various far-field super-resolution (SR) microscopy imaging methods using evanescent waves to transfer the high-frequency information of samples to the low-frequency passband of optical microscopy. Optimization algorithms are developed to reconstruct a SR image of the sample by utilizing the high-frequency information. These techniques can be collectively referred to as spatial-frequency-shift (SFS) SR microscopy. This review aims to summarize the basic principle of SR microscopy using evanescent illumination and introduce the advances in this research area. Some current challenges and possible directions are also discussed.
Reducing Heavy Metal Contamination in Soil and Water Using Phytoremediation
Authors: Zhakypbek Yryszhan , Kossalbayev Bekzhan D. , Belkozhayev Ayaz M. , Murat Toktar , Tursbekov Serik , Abdalimov Elaman , Pashkovskiy Pavel , Kreslavski Vladimir , Kuznetsov Vladimir , Allakhverdiev Suleyman I.
Published: June 2024

The increase in industrialization has led to an exponential increase in heavy metal (HM) soil contamination, which poses a serious threat to public health and ecosystem stability. This review emphasizes the urgent need to develop innovative technologies for the environmental remediation of intensive anthropogenic pollution. Phytoremediation is a sustainable and cost-effective approach for the detoxification of contaminated soils using various plant species. This review discusses in detail the basic principles of phytoremediation and emphasizes its ecological advantages over other methods for cleaning contaminated areas and its technical viability. Much attention has been given to the selection of hyperaccumulator plants for phytoremediation that can grow on heavy metal-contaminated soils, and the biochemical mechanisms that allow these plants to isolate, detoxify, and accumulate heavy metals are discussed in detail. The novelty of our study lies in reviewing the mechanisms of plant–microorganism interactions that greatly enhance the efficiency of phytoremediation as well as in discussing genetic modifications that could revolutionize the cleanup of contaminated soils. Moreover, this manuscript discusses potential applications of phytoremediation beyond soil detoxification, including its role in bioenergy production and biodiversity restoration in degraded habitats. This review concludes by listing the serious problems that result from anthropogenic environmental pollution that future generations still need to overcome and suggests promising research directions in which the integration of nano- and biotechnology will play an important role in enhancing the effectiveness of phytoremediation. These contributions are critical for environmental scientists, policy makers, and practitioners seeking to utilize phytoremediation to maintain the ecological stability of the environment and its restoration.
Identification of Multiple Diseases in Apple Leaf Based on Optimized Lightweight Convolutional Neural Network
Authors: Wang Bin , Yang Hua , Zhang Shujuan , Li Lili
Published: June 2024

In this study, our aim is to find an effective method to solve the problem of disease similarity caused by multiple diseases occurring on the same leaf. This study proposes the use of an optimized RegNet model to identify seven common apple leaf diseases. We conducted comparisons and analyses on the impact of various factors, such as training methods, data expansion methods, optimizer selection, image background, and other factors, on model performance. The findings suggest that utilizing offline expansion and transfer learning to fine-tune all layer parameters can enhance the model’s classification performance, while complex image backgrounds significantly influence model performance. Additionally, the optimized RegNet network model demonstrates good generalization ability for both datasets, achieving testing accuracies of 93.85% and 99.23%, respectively. These results highlight the potential of the optimized RegNet network model to achieve high-precision identification of different diseases on the same apple leaf under complex field backgrounds. This will be of great significance for intelligent disease identification in apple orchards in the future.
UV-B Radiation Disrupts Membrane Lipid Organization and Suppresses Protein Mobility of GmNARK in Arabidopsis
Authors: Liu Qiulin , Wang Tianyu , Ke Meiyu , Qian Chongzhen , Li Jiejie , Huang Xi , Gao Zhen , Chen Xu , Tu Tianli
Published: June 2024

While it is well known that plants interpret UV-B as an environmental cue and a potential stressor influencing their growth and development, the specific effects of UV-B-induced oxidative stress on the dynamics of membrane lipids and proteins remain underexplored. Here, we demonstrate that UV-B exposure notably increases the formation of ordered lipid domains on the plasma membrane (PM) and significantly alters the behavior of the Glycine max nodule autoregulation receptor kinase (GmNARK) protein in Arabidopsis leaves. The GmNARK protein was located on the PM and accumulated as small particles in the cytoplasm. We found that UV-B irradiation interrupted the lateral diffusion of GmNARK proteins on the PM. Furthermore, UV-B light decreases the efficiency of surface molecule internalization by clathrin-mediated endocytosis (CME). In brief, UV-B irradiation increased the proportion of the ordered lipid phase and disrupted clathrin-dependent endocytosis; thus, the endocytic trafficking and lateral mobility of GmNARK protein on the plasma membrane are crucial for nodule formation tuning. Our results revealed a novel role of low-intensity UV-B stress in altering the organization of the plasma membrane and the dynamics of membrane-associated proteins.
Catha edulis Leaves: Morphological Characterization and Anti-Inflammatory Properties in an In Vitro Model of Gastritis
Authors: Magnavacca Andrea , Giuliani Claudia , Roda Gabriella , Piazza Stefano , Martinelli Giulia , Pozzoli Carola , Maranta Nicole , Papini Alessio , Bottoni Martina , Casagni Eleonora , Dei Cas Michele , Fico Gelsomina , Dell’Agli Mario , Sangiovanni Enrico
Published: June 2024

Khat leaves, indigenous to eastern Africa, have been chewed for centuries for their stimulant effects, attributed to alkaloids such as cathinone and cathine. Although associated with gastric disorders like gastritis and gastro-oesophageal reflux disease, the underlying molecular mechanisms remain unclear. This study aimed to examine the morpho-anatomy of khat leaves using light microscopy and histochemistry and to assess the effects of leaf extracts and alkaloids on human gastric epithelial cells (GES-1). The study identified specific cells in the palisade–spongy transition zone as storage sites for psychoactive alkaloids. Leaf extracts were prepared by mimicking the chewing process, including a prolonged salivary phase followed by a gastric phase. Cytotoxicity and cell viability were evaluated using LDH and MTT assays, respectively. Additionally, the impact on IL-8 secretion, a key chemokine in gastric inflammation, was analysed under normal and TNF-α-stimulated conditions. The results showed no increase in cytotoxicity up to 250 µg/mL. However, there was a significant decrease in cell metabolism and a reduction in both basal and TNF-α-induced IL-8 secretion, but cathinone and cathine were inactive. These findings suggest that khat may not directly cause the gastric issues reported in the literature, which would rather be attributed to other confounding factors, highlighting the need for further research to clarify its biological impacts.
Transgenic Soybean for Production of Thermostable α-Amylase
Authors: Cao Zhenyan , Jiang Ye , Li Jiajie , Zheng Ting , Lin Chaoyang , Shen Zhicheng
Published: June 2024

Alpha-amylases are crucial hydrolase enzymes which have been widely used in food, feed, fermentation, and pharmaceutical industries. Methods for low-cost production of α-amylases are highly desirable. Soybean seed, functioning as a bioreactor, offers an excellent platform for the mass production of recombinant proteins for its ability to synthesize substantial quantities of proteins. In this study, we generated and characterized transgenic soybeans expressing the α-amylase AmyS from Bacillus stearothermophilus. The α-amylase expression cassettes were constructed for seed specific expression by utilizing the promoters of three different soybean storage peptides and transformed into soybean via Agrobacterium-mediated transformation. The event with the highest amylase activity reached 601 U/mg of seed flour (one unit is defined as the amount of enzyme that generates 1 micromole reducing ends per min from starch at 65 °C in pH 5.5 sodium acetate buffer). The optimum pH, optimum temperature, and the enzymatic kinetics of the soybean expressed enzyme are similar to that of the E. coli expressed enzyme. However, the soybean expressed α-amylase is glycosylated, exhibiting enhanced thermostability and storage stability. Soybean AmyS retains over 80% activity after 100 min at 75 °C, and the transgenic seeds exhibit no significant activity loss after one year of storage at room temperature. The accumulated AmyS in the transgenic seeds represents approximately 15% of the total seed protein, or about 4% of the dry seed weight. The specific activity of the transgenic soybean seed flour is comparable to many commercial α-amylase enzyme products in current markets, suggesting that the soybean flour may be directly used for various applications without the need for extraction and purification.
Ecological Niche Differentiation and Response to Climate Change of the African Endemic Family Myrothamnaceae
Authors: Wan Qisong , Du Shenglan , Chen Yu , Li Feng , Salah Radwa , Njenga Maxwell Njoroge , Li Jitao , Wang Shengwei
Published: June 2024

Studying the ecological niches of species and their responses to climate change can provide better conservation strategies for these species. Myrothamnaceae is endemic to Africa, comprising only two species that belong to Myrothamnus (M. flabellifolius and M. moschatus). These closely related species exhibit allopatric distributions, positioning them as ideal materials for studying the species ecological adaptation. This study explores the ecological niche differentiation between M. flabellifolius and M. moschatus and their response capabilities to future climate change. The results indicate that M. flabellifolius and M. moschatus have undergone niche differentiation. The main drivers of niche differences are the minimum temperature of the coldest month (Bio6) for M. flabellifolius, precipitation of the driest month (Bio14), and precipitation of the coldest quarter (Bio19) for M. moschatus. M. flabellifolius demonstrated a stronger adaptation to environments characterized by lower precipitation, relatively lower temperatures, and greater annual temperature variations compared to M. moschatus. Under future climate scenarios (SSP5-8.5, 2081–2100 years), the results show that approximately 85% of the total suitable habitat for M. flabellifolius will be lost, with an 85% reduction in high-suitability areas and almost complete loss of the original mid-low suitability areas. Concurrently, about 29% of the total suitable habitat for M. moschatus will be lost, with a 34% reduction in high suitability areas and roughly 60% of the original mid-low suitability areas becoming unsuitable. This suggests that M. flabellifolius will face greater threats under future climate change. This study contributes novel insight into niche differentiation in Myrothamnaceae and provides useful information for the conservation of this distinctive African lineage.