bluuebunny/crossref_metadata_2025 huggingface dataset:
Beta-Lactoglobulin as a Model Food Protein: How to Promote, Prevent, and Exploit Its Unfolding Processes
Authors: Barbiroli Alberto , Iametti Stefania , Bonomi Francesco
Published: February 2022

Bovine milk beta-lactoglobulin (BLG) is a small whey protein that is a common ingredient in many foods. Many of the properties of BLG relevant to the food industry are related to its unfolding processes induced by physical or chemical treatments. Unfolding occurs through a number of individual steps, generating transient intermediates through reversible and irreversible modifications. The rate of formation of these intermediates and of their further evolution into different structures often dictates the outcome of a given process. This report addresses the main structural features of the BLG unfolding intermediates under conditions that may facilitate or impair their formation in response to chemical or physical denaturing agents. In consideration of the short lifespan of the transient species generated upon unfolding, this review also discusses how various methodological approaches may be adapted in exploring the process-dependent structural modifications of BLG from a kinetic and/or a thermodynamic standpoint. Some of the conceptual and methodological approaches presented and discussed in this review can provide hints for improving the understanding of transient conformers formation by proteins present in other food systems, as well as when other physical or chemical denaturing agents are acting on proteins much different from BLG in complex food systems.
Supercritical CO2 Plant Extracts Show Antifungal Activities against Crop-Borne Fungi
Authors: Schoss Katja , Kočevar Glavač Nina , Dolenc Koce Jasna , Anžlovar Sabina
Published: February 2022

Fungal infections of cultivated food crops result in extensive losses of crops at the global level, while resistance to antifungal agents continues to grow. Supercritical fluid extraction using CO2 (SFE-CO2) has gained attention as an environmentally well-accepted extraction method, as CO2 is a non-toxic, inert and available solvent, and the extracts obtained are, chemically, of greater or different complexities compared to those of conventional extracts. The SFE-CO2 extracts of Achillea millefolium, Calendula officinalis, Chamomilla recutita, Helichrysum arenarium, Humulus lupulus, Taraxacum officinale, Juniperus communis, Hypericum perforatum, Nepeta cataria, Crataegus sp. and Sambucus nigra were studied in terms of their compositions and antifungal activities against the wheat- and buckwheat-borne fungi Alternaria alternata, Epicoccum nigrum, Botrytis cinerea, Fusarium oxysporum and Fusarium poae. The C. recutita and H. arenarium extracts were the most efficacious, and these inhibited the growth of most of the fungi by 80% to 100%. Among the fungal species, B. cinerea was the most susceptible to the treatments with the SFE-CO2 extracts, while Fusarium spp. were the least. This study shows that some of these SFE-CO2 extracts have promising potential for use as antifungal agents for selected crop-borne fungi.
State of the Art on Green Route Synthesis of Gold/Silver Bimetallic Nanoparticles
Authors: Scala Angela , Neri Giulia , Micale Nicola , Cordaro Massimiliano , Piperno Anna
Published: February 2022

Recently, bimetallic nanoparticles (BMNPs) blending the properties of two metals in one nanostructured system have generated enormous interest due to their potential applications in various fields including biosensing, imaging, nanomedicine, and catalysis. BMNPs have been developed later with respect to the monometallic nanoparticles (MNPs) and their physicochemical and biological properties have not yet been comprehensively explored. The manuscript aims at collecting the main design criteria used to synthetize BMNPs focusing on green route synthesis. The influence of experimental parameters such as temperature, time, reagent concentrations, capping agents on the particle growth and colloidal stability are examined. Finally, an overview of their nanotechnological applications and biological profile are presented.
π–π Stacking Interaction of Metal Phenoxyl Radical Complexes
Authors: Oshita Hiromi , Shimazaki Yuichi
Published: February 2022

π–π stacking interaction is well-known to be one of the weak interactions. Its importance in the stabilization of protein structures and functionalization has been reported for various systems. We have focused on a single copper oxidase, galactose oxidase, which has the π–π stacking interaction of the alkylthio-substituted phenoxyl radical with the indole ring of the proximal tryptophan residue and catalyzes primary alcohol oxidation to give the corresponding aldehyde. This stacking interaction has been considered to stabilize the alkylthio-phenoxyl radical, but further details of the interaction are still unclear. In this review, we discuss the effect of the π–π stacking interaction of the alkylthio-substituted phenoxyl radical with an indole ring.
Antioxidant and Antimicrobial Activities of Chemically-Characterized Essential Oil from Artemisia aragonensis Lam. against Drug-Resistant Microbes
Authors: Chebbac Khalid , Ghneim Hazem K. , El Moussaoui Abdelfattah , Bourhia Mohammed , El Barnossi Azeddin , Benziane Ouaritini Zineb , Salamatullah Ahmad Mohammad , Alzahrani Abdulhakeem , Aboul-Soud Mourad A. M. , Giesy John P. , Guemmouh Raja
Published: February 2022

This study investigated the chemical composition, antioxidant and antimicrobial activity of essential oil extracted from Artemisia aragonensis Lam. (EOA). Hydrodistillation was employed to extract EOA. Gas chromatography with flame ionization detection (GC-FID) and gas chromatography-mass spectrometry analyses (GC-MS) were used to determine the phytochemical composition of EOA. Antioxidant potential was examined in vitro by use of three tests: 2.2-diphenyl-1-picrilhidrazil (DPPH), ferric reducing activity power (FRAP) and total antioxidant capacity assay (TAC). Agar diffusion and microdilution bioassays were used to assess antimicrobial activity. GC/MS and GC-FID detected 34 constituents in the studied EOA. The major component was Camphor (24.97%) followed by Borneol (13.20%), 1,8 Cineol (10.88%), and Artemisia alcohol (10.20%). EOA exhibited significant antioxidant activity as measured by DPPH and FRAP assays, with IC50 and EC50 values of 0.034 ± 0.004 and 0.118 ± 0.008 mg/mL, respectively. EOA exhibited total antioxidant capacity of 7.299 ± 1.774 mg EAA/g. EOA exhibited potent antibacterial activity as judged by the low minimum inhibitory concentration (MIC) values against selected clinically-important pathogenic bacteria. MIC values of 6.568 ± 1.033, 5.971 ± 1.033, 7.164 ± 0.0 and 5.375 ± 0.0 μg/mL were observed against S. aureus, B. subtills, E. coli 97 and E. coli 57, respectively. EOA displayed significant antifungal activity against four strains of fungi: F. oxysporum, C. albicans, A. flavus and A. niger with values of 21.50 ± 0.43, 5.31 ± 0.10, 21.50 ± 0.46 and 5.30 ± 0.036 μg/mL, respectively. The results of the current study highlight the importance of EOA as an alternative source of natural antioxidant and antibacterial drugs to combat antibiotic-resistant microbes and free radicals implicated in the inflammatory responses accompanying microbial infection.
Evaluation of Fast and Sensitive Proteome Profiling of FF and FFPE Kidney Patient Tissues
Authors: Dapic Irena , Uwugiaren Naomi , Kers Jesper , Mohammed Yassene , Goodlett David R. , Corthals Garry
Published: February 2022

The application of proteomics to fresh frozen (FF) and formalin-fixed paraffin-embedded (FFPE) human tissues is an important development spurred on by requests from stakeholder groups in clinical fields. One objective is to complement current diagnostic methods with new specific molecular information. An important goal is to achieve adequate and consistent protein recovery across and within large-scale studies. Here, we describe development of several protocols incorporating mass spectrometry compatible detergents, including Rapigest, PPS, and ProteaseMax. Methods were applied on 4 and 15 μm thick FF tissues, and 4 μm thick FFPE tissues. We evaluated sensitivity and repeatability of the methods and found that the protocol containing Rapigest enabled detection of 630 proteins from FF tissue of 1 mm2 and 15 μm thick, whereas 498 and 297 proteins were detected with the protocols containing ProteaseMax and PPS, respectively. Surprisingly, PPS-containing buffer showed good extraction of the proteins from 4 μm thick FFPE tissue with the average of 270 protein identifications (1 mm2), similar to the results on 4 μm thick FF. Moreover, we found that temperature increases during incubation with urea on 4 μm thick FF tissue revealed a decrease in the number of identified proteins and increase in the number of the carbamylated peptides.
A Comprehensive Investigation of the Structural, Thermal, and Biological Properties of Fully Randomized Biomedical Polyesters Synthesized with a Nontoxic Bismuth(III) Catalyst
Authors: Domańska Izabela M. , Zgadzaj Anna , Kowalczyk Sebastian , Zalewska Aldona , Oledzka Ewa , Cieśla Krystyna , Plichta Andrzej , Sobczak Marcin
Published: February 2022

Aliphatic polyesters are the most common type of biodegradable synthetic polymer used in many pharmaceutical applications nowadays. This report describes the ring-opening polymerization (ROP) of l-lactide (L-LA), ε-caprolactone (CL) and glycolide (Gly) in the presence of a simple, inexpensive and convenient PEG200-BiOct3 catalytic system. The chemical structures of the obtained copolymers were characterized by 1H- or 13C-NMR. GPC was used to estimate the average molecular weight of the resulting polyesters, whereas TGA and DSC were employed to determine the thermal properties of polymeric products. The effects of temperature, reaction time, and catalyst content on the polymerization process were investigated. Importantly, the obtained polyesters were not cyto- or genotoxic, which is significant in terms of the potential for medical applications (e.g., for drug delivery systems). As a result of transesterification, the copolymers obtained had a random distribution of comonomer units along the polymer chain. The thermal analysis indicated an amorphous nature of poly(l-lactide-co-ε-caprolactone) (PLACL) and a low degree of crystallinity of poly(ε-caprolactone-co-glycolide) (PCLGA, Xc = 15.1%), in accordance with the microstructures with random distributions and short sequences of comonomer units (l = 1.02–2.82). Significant differences in reactivity were observed among comonomers, confirming preferential ring opening of L-LA during the copolymerization process.
Flavonoids: Antiplatelet Effect as Inhibitors of COX-1
Authors: Zaragozá Cristina , Álvarez-Mon Miguel Ángel , Zaragozá Francisco , Villaescusa Lucinda
Published: February 2022

Flavonoids are compounds with a benzopyranic structure that exhibits multiple pharmacological activities. They are known for their venotonic activity, but their mechanism of action remains unclear. It is thought that, as this mechanism is mediated by prostaglandins, these compounds may interfere with the arachidonic acid (AA) cascade. These assays are designed to measure the antiplatelet aggregation capacity of quercetin, rutin, diosmetin, diosmin, and hidrosmin, as well as to evaluate a potential structure−activity ratio. In this paper, several studies on platelet aggregation at different concentrations (from 0.33 mM to 1.5 mM) of different flavone compounds are conducted, measuring platelet aggregation by impedance aggregometry, and the cyclooxygenase (COX) activity by metabolites generated, including the activity of the pure recombinant enzyme in the presence of these polyphenols. The results obtained showed that quercetin and diosmetin aglycones have a greater antiplatelet effect and inhibit the COX enzyme activity to a greater extent than their heterosides; however, the fact that greater inhibition of the pure recombinant enzyme was achieved by heterosides suggests that these compounds may have difficulty in crossing biological membranes. In any case, in view of the results obtained, it can be concluded that flavonoids could be useful as coadjuvants in the treatment of cardiovascular pathologies.
Alpha B-Crystallin in Muscle Disease Prevention: The Role of Physical Activity
Authors: Dimauro Ivan , Caporossi Daniela
Published: February 2022

HSPB5 or alpha B-crystallin (CRYAB), originally identified as lens protein, is one of the most widespread and represented of the human small heat shock proteins (sHSPs). It is greatly expressed in tissue with high rates of oxidative metabolism, such as skeletal and cardiac muscles, where HSPB5 dysfunction is associated with a plethora of human diseases. Since HSPB5 has a major role in protecting muscle tissues from the alterations of protein stability (i.e., microfilaments, microtubules, and intermediate filament components), it is not surprising that this sHSP is specifically modulated by exercise. Considering the robust content and the protective function of HSPB5 in striated muscle tissues, as well as its specific response to muscle contraction, it is then realistic to predict a specific role for exercise-induced modulation of HSPB5 in the prevention of muscle diseases caused by protein misfolding. After offering an overview of the current knowledge on HSPB5 structure and function in muscle, this review aims to introduce the reader to the capacity that different exercise modalities have to induce and/or activate HSPB5 to levels sufficient to confer protection, with the potential to prevent or delay skeletal and cardiac muscle disorders.
Mintaimycins, a Group of Novel Peptide Metabolites from Micromonospora sp. C-3509
Authors: Hu Xiaomin , Wang Ying , Zhao Chunyan , Li Shufen , Hu Xinxin , You Xuefu , Shen Jiajia , Wang Zhen , Hong Bin , Jiang Bingya , Du Yu , Wu Linzhuan
Published: February 2022

A group of peptide metabolites (1–4), designated as mintaimycins, were isolated from Micromonospora sp. C-3509. The planar structures of mintaimycins were determined by combination of mass spectrometry, 1D and 2D NMR spectroscopy, and the stereochemistry of mintaimycins were partially resolved by Marfey’s or Mosher’s method. Mintaimycins featured a central β-methylphenylalanine or phenylalanine linked at its amino group with 5-methyl-2-hexenoic acid, and at its carboxyl group with 5-hydroxy-norleucine or leucine that combined a derivative of hexanoic acid or 4-methylpentanoic acid. Mintaimycin A1 (1), the principal component, was found to exhibit the biological activity of inducing pre-adipocyte differentiation of 3T3-L1 fibroblast cells at 10.0 μmol/L.
The Paradox of the Mediterranean Diet in Pediatric Age during the COVID-19 Pandemic
Authors: Verduci Elvira , Fiore Giulia , Di Profio Elisabetta , Zuccotti Gian Vincenzo
Published: February 2022

The outbreak of the COVID-19 pandemic, whose causative agent is Severe Acute Respiratory Syndrome Coronavirus 2, has caused a global crisis that has had a major impact on the health of the global population [...]
Co-Administration of Iron and Bioavailable Curcumin Reduces Levels of Systemic Markers of Inflammation and Oxidative Stress in a Placebo-Controlled Randomised Study
Authors: Tiekou Lorinczova Helena , Begum Gulshanara , Temouri Lina , Renshaw Derek , Zariwala Mohammed Gulrez
Published: February 2022

Ferrous sulphate (FS) is widely used as an iron supplement to treat iron deficiency (ID), but is known to induce inflammation causing gastric side-effects resulting in poor adherence to supplement regimens. Curcumin, a potent antioxidant, has been reported to suppress inflammation via down regulation of NF-κB. The aim of the present double blind, placebo-controlled randomised trial was to assess whether co-administration of FS with a formulated, bioavailable form of curcumin (HydroCurc™) could reduce systemic inflammation and/or gastrointestinal side-effects. This study recruited 155 healthy participants (79 males; 26.42 years ± 0.55 and 76 females; 25.82 years ± 0.54), randomly allocated to one of five different treatment groups: iron and curcumin placebo (FS0_Plac), low dose (18 mg) iron and curcumin placebo (FS18_Plac), low dose iron and curcumin (FS18_Curc), high dose (65 mg) iron and curcumin placebo (FS65_Plac), and high dose iron and curcumin (FS65_Curc). Completed questionnaires and blood samples were collected from all participants at baseline (day 1), mid-point (day 21), and at end-point (day 42). Results showed a significant reduction in IL-6 in the FS65_Curc group (0.06 pg/mL ± 0.02, p = 0.0073) between the mid-point and end-point. There was also a significant reduction in mean plasma TNF levels in the FS65_Curc (0.65 pg/mL ± 0.17, p = 0.0018), FS65_Plac (0.39 pg/mL ± 0.15, p = 0.0363), and FS18_Curc (0.35 pg/mL ± 0.13, p = 0.0288) groups from mid-point to end-point. A significant increase was observed in mean plasma TBARS levels (0.10 µM ± 0.04, p = 0.0283) in the F18_Plac group from baseline to end-point. There was a significant association with darker stools between FS0_Plac vs. FS65_Plac (p = 0.002, Fisher’s exact test) suggesting that high iron dose in the absence of curcumin leads to darker stools. A reduction in inflammation-related markers in response to co-administering supplemental iron alongside formulated curcumin suggests a reduction in systemic inflammation. This supplementation approach may therefore be a more cost effective and convenient alternative to current oral iron-related treatments, with further research to be conducted.
Choline Kinetics in Neonatal Liver, Brain and Lung—Lessons from a Rodent Model for Neonatal Care
Authors: Bernhard Wolfgang , Raith Marco , Shunova Anna , Lorenz Stephan , Böckmann Katrin , Minarski Michaela , Poets Christian F. , Franz Axel R.
Published: February 2022

Choline requirements are high in the rapidly growing fetus and preterm infant, mainly serving phosphatidylcholine (PC) synthesis for parenchymal growth and one-carbon metabolism via betaine. However, choline metabolism in critical organs during rapid growth is poorly understood. Therefore, we investigated the kinetics of D9-choline and its metabolites in the liver, plasma, brain and lung in 14 d old rats. Animals were intraperitoneally injected with 50 mg/kg D9-choline chloride and sacrificed after 1.5 h, 6 h and 24 h. Liver, plasma, lungs, cerebrum and cerebellum were analyzed for D9-choline metabolites, using tandem mass spectrometry. In target organs, D9-PC and D9-betaine comprised 15.1 ± 1.3% and 9.9 ± 1.2% of applied D9-choline at 1.5 h. D9-PC peaked at 1.5 h in all organs, and decreased from 1.5–6 h in the liver and lung, but not in the brain. Whereas D9-labeled PC precursors were virtually absent beyond 6 h, D9-PC increased in the brain and lung from 6 h to 24 h (9- and 2.5-fold, respectively) at the expense of the liver, suggesting PC uptake from the liver via plasma rather than local synthesis. Kinetics of D9-PC sub-groups suggested preferential hepatic secretion of linoleoyl-PC and acyl remodeling in target organs. D9-betaine showed rapid turnover and served low-level endogenous (D3-)choline synthesis. In conclusion, in neonatal rats, exogenous choline is rapidly metabolized to PC by all organs. The liver supplies the brain and lung directly with PC, followed by organotypic acyl remodeling. A major fraction of choline is converted to betaine, feeding the one-carbon pool and this must be taken into account when calculating choline requirements.
Gut Seasons: Photoperiod Effects on Fecal Microbiota in Healthy and Cafeteria-Induced Obese Fisher 344 Rats
Authors: Arreaza-Gil Verónica , Escobar-Martínez Iván , Suárez Manuel , Bravo Francisca , Muguerza Begoña , Arola-Arnal Anna , Torres-Fuentes Cristina
Published: February 2022

Gut microbiota and biological rhythms are emerging as key factors in the modulation of several physiological and metabolic processes. However, little is known about their interaction and how this may affect host physiology and metabolism. Several studies have shown oscillations of gut microbiota that follows a circadian rhythmicity, but, in contrast, variations due to seasonal rhythms have not been sufficiently investigated yet. Thus, the goal of this study was to investigate the impact of different photoperiods, which mimic seasonal changes, on fecal microbiota composition and how this interaction affects diet-induced obesity development. To this aim, Fisher 344 male rats were housed under three photoperiods (L6, L12 and L18) and fed with standard chow diet or cafeteria diet (CAF) for 9 weeks. The 16S ribosomal sequencing of collected fecal samples was performed. The photoperiod exposure significantly altered the fecal microbiota composition under L18, especially in CAF-fed rats. Moreover, these alterations were associated with changes in body weight gain and different fat parameters. These findings suggest a clear impact of seasonal rhythms on gut microbiota, which ultimately translates into different susceptibilities to diet-induced obesity development. This is the first time to our knowledge that the photoperiod impact on gut microbiota composition has been described in an obesity context although further studies are needed in order to elucidate the mechanisms involved.
Comment on Depoorter, L.; Vandenplas, Y. Probiotics in Pediatrics. A Review and Practical Guide. Nutrients 2021, 13, 2176
Authors: von Bünau Rudolf , Erhardt Andreas , Stange Eduard
Published: February 2022

We read the review by Depoorter et al. [...]
Patient Reported Experience Measure in Endoscopic Ultrasonography: The PREUS Study Protocol
Authors: Apadula Laura , Capurso Gabriele , Ambrosi Alessandro , Arcidiacono Paolo Giorgio
Published: February 2022

The evaluation of the patient’s experience is becoming increasingly important as a better patient experience can improve the quality of the health service delivered. Patient-reported experience measures (PREMs) are self-report assessment tools provided to patients about their experience during any health event. There are few PREM instruments in the field of gastrointestinal endoscopy, and none is specific for endoscopic ultrasound (EUS). This study aims to develop a questionnaire to evaluate the experience of patients undergoing EUS, identifying and prioritizing the factors related to the patient’s experience. The study will consist of several phases: (A) tool creation; (B) face and content validity; (C) ranking: to evaluate the relevance of the identified questions in the previous phase; (D) questionnaire creation and validity testing. The final output will be the production of a specific tool that can be used to measure patients’ experience during EUS. This questionnaire may become a relevant part of actions taken to measure the quality of care provided to patients undergoing EUS. Furthermore, correlation between health care providers’ and patients’ views of the relevance of the included items will allow optimization of empathetic and psychological aspects.
Health Plans for Suicide Prevention in Spain: A Descriptive Analysis of the Published Documents
Authors: Sufrate-Sorzano Teresa , Jiménez-Ramón Elena , Garrote-Cámara María Elena , Gea-Caballero Vicente , Durante Angela , Júarez-Vela Raúl , Santolalla-Arnedo Iván
Published: February 2022

The number of deaths by suicide worldwide each year is more than 800,000 people, which is equivalent to one death every 40 seconds. Suicide prevention has been listed by the World Health Organisation as a global imperative and has become a priority for global public health. This descriptive study describes and compares the intervention components included in the suicide prevention plans in the different provinces of Spain. We analysed the published documents through an extensive literature search and summarised the findings using descriptive content analysis. The search was carried out through the official websites of the government and health departments of each province in addition to consulting other official digital platforms such as the National Suicide Observatory, the World Health Organisation and the National Institute of Statistics. The results show the most relevant differences between the prevention plans, revealing that although all the activities included were related to the health sector, not all of them include prevention aimed at the general population level. We conclude that there is a lack of interventions related to the application of universal prevention, while selective and indicated prevention are the most developed tools in Spain.
Anti-Inflammatory and Neuroprotective Effect of the Anti-Obesity Dietary Supplement Dekosilhue® in an In Vitro Model of Neuroinflammation
Authors: Borgonetti Vittoria , Cenci Lorenzo , Galeotti Nicoletta
Published: February 2022

Obesity is now recognized as a global health problem and has reached epidemic proportions, affecting both developed and developing countries. This condition is associated with an increased incidence in central nervous system (CNS) disorders. The intake of foods or supplements with anti-inflammatory activity could potentially lead to a lower impact of obesity on the functionality of CNS. Therefore, in this work we tested the effect of a dietary supplement, DEKOSILHUE® (DSK), containing polyphenols enriched herbal extracts and successfully used for improving the control of body weight, in an in vitro inflammatory model. LPS-stimulated immortalized microglia BV2 cells have been used as a model of neuroinflammation. The stimulation with LPS increased the release and activation of several neuroinflammatory mediators, such as IKB⍺, HDAC-1, and IL-1ß. DSK reduced the levels and the activation of all these pro-inflammatory markers. In addition, DSK increased the expression of the anti-inflammatory cytokine IL-10, promoting an anti-inflammatory response. Finally, we showed that the anti-inflammatory effect of DSK can prevent the neurotoxicity related to microglia hyperactivation. In conclusion, DSK could potentially help in controlling neuroinflammation CNS, reducing microglia pro-inflammatory activation.
Diagnostic Considerations for Non-Acanthamoeba Amoebic Keratitis and Clinical Outcomes
Authors: Moran Siobhan , Mooney Ronnie , Henriquez Fiona L.
Published: February 2022

Cases of amoebic keratitis involving species other than Acanthamoeba are hypothesised to be underdiagnosed and poorly understood. Amoebic keratitis is debilitating and associated with chronic visual impairment. Understanding associated symptoms of non-Acanthamoeba amoebic keratitis could facilitate new diagnostic procedures and enable prompt treatment, ultimately leading to improved patient outcomes. Thus, a review of the literature was undertaken surrounding non-Acanthamoeba amoebic keratitis. Cases were geographically widespread and mostly confined to contact lens wearers ≤ 30 years old exposed to contaminated water sources and/or demonstrating poor lens hygiene. Vermamoeba vermiformis (previously Hartmanella vermiformis) was the most common causative agent, and a moderate number of mixed keratitis cases were also reported. A crucial disease indicator was early onset stromal deterioration/ulcerations, reported in 10 of the studies, usually only occurring in advanced Acanthamoeba keratitis. Mixed infections were the most difficult to treat, often requiring keratoplasty after unsuccessful combination treatment regimens. New diagnostic measures for non-Acanthamoeba amoebic keratitis should consider early onset stromal disease as a key disease indicator. Deep corneal scrapes are also necessary for accurate amoebic identification. Moreover, a combination approach to diagnosis is advised and should involve culture, microscopy and PCR techniques. In vitro drug sensitivity tests should also be conducted to help develop patient-specific treatment regimes.
Evaluation of Betacoronavirus OC43 and SARS-CoV-2 Elimination by Zefero Air Sanitizer Device in a Novel Laboratory Recirculation System
Authors: Nicolò Marco Sebastiano , Rizzo Maria Giovanna , Palermo Nicoletta , Gugliandolo Concetta , Cuzzocrea Salvatore , Guglielmino Salvatore P. P.
Published: February 2022

Indoor air sanitizers contrast airborne diseases and particularly severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)/Coronavirus disease 2019 (COVID-19). The commercial air sanitizer Zefero (Cf7 S.r.l., San Giovanni La Punta, Italy) works alternatively using a set of integrated disinfecting technologies (namely Photocatalysis/UV mode) or by generating ozone (Ozone mode). Here we evaluated the virucidal efficacy of Zefero setup modes against human Betacoronavirus OC43 and SARS-CoV-2. For this purpose, we designed a laboratory test system in which each virus, as aerosol, was treated with Photocatalysis/UV or Ozone mode and returned into a recirculation plexiglass chamber. Aerosol samples were collected after different times of exposure, corresponding to different volumes of air treated. The viral RNA concentration was determined by qRT-PCR. In Photocatalysis/UV mode, viral RNA of OC43 or SARS-CoV-2 was not detected after 120 or 90 min treatment, respectively, whereas in Ozone mode, viruses were eliminated after 30 or 45 min, respectively. Our results indicated that the integrated technologies used in the air sanitizer Zefero are effective in eliminating both viruses. As a reliable experimental system, the recirculation chamber developed in this study represents a suitable apparatus for effectively comparing the disinfection capacity of different air sanitizers.
Babesia and Theileria Identification in Adult Ixodid Ticks from Tapada Nature Reserve, Portugal
Authors: Fernández Nélida , Revuelta Belen , Aguilar Irene , Soares Jorge Francisco , Zintl Annetta , Gray Jeremy , Montero Estrella , Gonzalez Luis Miguel
Published: February 2022

This study, conducted in a nature reserve in southern Portugal, investigated the frequency and diversity of tick-borne piroplasms in six species of adult ixodid ticks removed from 71 fallow deer (Dama dama) and 12 red deer (Cervus elaphus), collected over the period 2012–2019. The majority of 520 ticks were Ixodes ricinus (78.5%), followed by Rhipicephalus sanguineus sensu lato, Hyalomma lusitanicum, Haemaphysalis punctata, Dermacentor marginatus, and Ixodes hexagonus. The R. sanguineus ticks collected from the deer were clearly exophilic, in contrast to the endophilic species usually associated with dogs. Four tick-borne piroplasms, including Theileria spp., and the zoonotic species, Babesia divergens and Babesia microti, were detected. B. divergens 18S rDNA, identical to that of the bovine reference strain U16370 and to certain strains from red deer, was detected in I. ricinus ticks removed from fallow deer. The sporadic detection of infections in ticks removed from the same individual hosts suggests that the piroplasms were present in the ticks rather than the hosts. Theileria sp. OT3 was found in I. ricinus and, along with T. capreoli, was also detected in some of the other tick species. The natural vector and pathogenic significance of this piroplasm are unknown.
Particle Size Effect of Curcumin Nanocrystals on Transdermal and Transfollicular Penetration by Hyaluronic Acid-Dissolving Microneedle Delivery
Authors: Xiang Hong , Xu Sai , Li Jingyuan , Pan Shihui , Miao Xiaoqing
Published: February 2022

Microneedles are one promising penetration enhancement vehicle to overcome the stratum corneum skin barrier, which hampers the penetration of drug nanocrystals by transdermal delivery. In order to clarify the particle size effect of nanocrystals on transdermal delivery, 60 nm, 120 nm, and 480 nm curcumin nanocrystals were fabricated and incorporated into dissolving hyaluronic acid polysaccharide microneedles. The microneedles showed good mechanical strength with 1.4 N/needle, possessing the ability to insert into the skin. The passive permeation results showed that the smaller particle size of 60 nm curcumin nanocrystals diffused faster and deeper than the larger 120 nm and 480 nm curcumin nanocrystals with size-dependent diffusion behaviors. Thereafter, higher concentration gradients and overlap diffusional coronas also formed in the skin layers by the smaller-particle-size nanocrystals. Furthermore, the diffusion rate of the smaller particle size of curcumin nanocrystals to the hair follicle was also higher than that of the larger curcumin nanocrystals. In conclusion, the particle sizes of curcumin nanocrystals influenced the transdermal and transfollicular penetration in deeper skin layers
Long-Term Stability Prediction for Developability Assessment of Biopharmaceutics Using Advanced Kinetic Modeling
Authors: Evers Andreas , Clénet Didier , Pfeiffer-Marek Stefania
Published: February 2022

A crucial aspect of pharmaceutical development is the demonstration of long-term stability of the drug product. Biopharmaceuticals, such as proteins or peptides in liquid formulation, are typically administered via parental routes and should be stable over the shelf life, which generally includes a storing period (e.g., two years at 5 °C) and optionally an in-use period (e.g., 28 days at 30 °C). Herein, we present a case study where chemical degradation of SAR441255, a therapeutic peptide, in different formulations in combination with primary packaging materials was analyzed under accelerated conditions to derive long-term stability predictions for the recommended storing conditions (two years at 5 °C plus 28 days at 30 °C) using advanced kinetic modeling. These predictions served as a crucial decision parameter for the entry into clinical development. Comparison with analytical data measured under long-term conditions during the subsequent development phase demonstrated a high prediction accuracy. These predictions provided stability insights within weeks that would otherwise take years using measurements under long-term stability conditions only. To our knowledge, such in silico studies on stability predictions of a therapeutic peptide using accelerated chemical degradation data and advanced kinetic modeling with comparisons to subsequently measured real-life long-term stability data have not been described in literature before.
QbD Consideration for Developing a Double-Layered Tablet into a Single-Layered Tablet with Telmisartan and Amlodipine
Authors: Kim Joo-Eun , Park Young-Joon
Published: February 2022

The aim of this study was to develop a single-layered version of commercially available Twynstar® (Telmisartan + Amlodipine) double-layered tablets to improve the dosing convenience. A quality-by-design approach was applied to develop the single-layered version. To evaluate the range and cause of risks for a single-layered tablet in the formulation design research, we used the tools of the risk assessment, initial risk assessment of preliminary hazard analysis and main risk assessment of failure mode and effect analysis to determine the parameters affecting formulation, drug dissolution, and impurities. The critical material attributes were the stabilizer and disintegrant, and the critical process parameters were the wet granulation and tableting process. The optimal range of the design space was determined using the central composite design in the wet granulation and tablet compression processes. The stabilizer, kneading time, and disintegrant of the wet granulation were identified as X values affecting Y values. The compression force and turret speed in the tablet compression were identified as X values affecting Y values. After deciding on the design space with the deduced Y values, the single-layered tablets were formulated, and their dissolution patterns were compared with that of the double-layered tablet. The selected quality-by-design (QbD) approach single-layered tablet formulated using design space were found to be bioequivalent to the Twynstar® double-layered tablets. Hence, the development of single-layered tablets with two API using the QbD approach could improve the medication compliance of patients and could be used as a platform to overcome time-consuming and excessive costs and the technical and commercial limitations related to various multi-layered tablets.
Flexible Plenoptic X-ray Microscopy
Authors: Longo Elena , Alj Domenico , Batenburg Joost , de La Rochefoucauld Ombeline , Herzog Charlotte , Greving Imke , Li Ying , Lyubomirskiy Mikhail , Falch Ken Vidar , Estrela Patricia , Flenner Silja , Viganò Nicola , Fajardo Marta , Zeitoun Philippe
Published: February 2022

X-ray computed tomography (CT) is an invaluable technique for generating three-dimensional (3D) images of inert or living specimens. X-ray CT is used in many scientific, industrial, and societal fields. Compared to conventional 2D X-ray imaging, CT requires longer acquisition times because up to several thousand projections are required for reconstructing a single high-resolution 3D volume. Plenoptic imaging—an emerging technology in visible light field photography—highlights the potential of capturing quasi-3D information with a single exposure. Here, we show the first demonstration of a flexible plenoptic microscope operating with hard X-rays; it is used to computationally reconstruct images at different depths along the optical axis. The experimental results are consistent with the expected axial refocusing, precision, and spatial resolution. Thus, this proof-of-concept experiment opens the horizons to quasi-3D X-ray imaging, without sample rotation, with spatial resolution of a few hundred nanometres.
Cereal Germination under Low Oxygen: Molecular Processes
Authors: Gómez-Álvarez Eva María , Pucciariello Chiara
Published: February 2022

Cereal crops can differ greatly in tolerance to oxygen shortage under germination and seedling establishment. Rice is able to germinate and elongate the coleoptile under submergence and anoxia. This capacity has been attributed to the successful use of starchy reserves through a molecular pathway activated by sugar starvation and low oxygen. This pathway culminates with the expression of α-amylases to provide sugars that fuel the sink organs. On the contrary, barley and wheat are unable to germinate under anoxia. The sensitivity of barley and wheat is likely due to the incapacity to use starch during germination. This review highlights what is currently known about the molecular mechanisms associated with cereal germination and seedling establishment under oxygen shortage with a special focus on barley and rice. Insights into the molecular mechanisms that support rice germination under low oxygen and into those that are associated with barley sensitivity may be of help for genetic improvement programs.
Accumulation of Salicylic Acid and Related Metabolites in Selaginella moellendorffii
Authors: Berim Anna , Gang David R.
Published: February 2022

Salicylic acid (SA) is a phytohormone that plays manifold roles in plant growth, defense, and other aspects of plant physiology. The concentration of free SA in plants is fine-tuned by a variety of structural modifications. SA is produced by all land plants, yet it is not known whether its metabolism is conserved in all lineages. Selaginella moellendorffii is a lycophyte and thus a representative of an ancient clade of vascular plants. Here, we evaluated the accumulation of SA and related metabolites in aerial parts of S. moellendorffii. We found that SA is primarily stored as the 2-O-β-glucoside. Hydroxylated derivatives of SA are also produced by S. moellendorffii and stored as β-glycosides. A candidate signal for SA aspartate was also detected. Phenylpropanoic acids also occur in S. moellendorffii tissue. Only o-coumaric acid is stored as the β-glycoside, while caffeic, p-coumaric, and ferulic acids accumulate as alkali-labile conjugates. An in silico search for enzymes involved in conjugation and catabolism of SA in the S. moellendorffii genome indicated that experimental characterization is necessary to clarify the physiological functions of the putative orthologs. This study sheds light on SA metabolism in an ancestral plant species and suggests directions towards elucidating the underlying mechanisms.
Vicia–Micronucleus Test Application for Saline Irrigation Water Risk Assessment
Authors: Souguir Dalila , Berndtsson Ronny , Mzahma Sourour , Filali Hanen , Hachicha Mohamed
Published: February 2022

In view of climate change, increasing soil salinity is expected worldwide. It is therefore important to improve prediction ability of plant salinity effects. For this purpose, brackish/saline irrigation water from two areas in central and coastal Tunisia was sampled. The water samples were classified as C3 (EC: 2.01–2.24 dS m−1) and C4 (EC: 3.46–7.00 dS m−1), indicating that the water was questionable and not suitable for irrigation, respectively. The water samples were tested for their genotoxic potential and growth effects on Vicia faba seedlings. Results showed a decrease in mitotic index (MI) and, consequently, growth parameters concomitant to the appearance of micronucleus (MCN) and chromosome aberrations when the water salinity increased. Salt ion concentration had striking influence on genome stability and growth parameters. Pearson correlation underlined the negative connection between most ions in the water inappropriate for irrigation (C4) and MI as well as growth parameters. MI was strongly influenced by Mg2+, Na+, Cl−, and to a less degree Ca2+, K+, and SO42−. Growth parameters were moderately to weakly affected by K+ and Ca2+, respectively. Re-garding MCN, a very strong positive correlation was found for MCN and K+. Despite its short-term application, the Vicia-MCN Test showed a real ability to predict toxicity induced by salt ions confirming that is has a relevant role in hazard identification and risk assessment of salinity effects.
Thickness Dependence of Electronic Structure and Optical Properties of F8BT Thin Films
Authors: Ghasemi Bita , Ševčík Jakub , Nádaždy Vojtěch , Végsö Karol , Šiffalovič Peter , Urbánek Pavel , Kuřitka Ivo
Published: February 2022

Electronic devices based on polymer thin films have experienced a tremendous increase in their efficiency in the last two decades. One of the critical factors that affects the efficiency of polymer solar cells or light emitting devices is the presence of structural defects that controls non-radiative recombination. The purpose of this report is to demonstrate a non-trivial thickness dependence of optoelectronic properties and structure (dis)order in thin conductive poly(9,9-dioctyfluorene-alt-benzothiadiazole), F8BT, polymer films. The UV-Vis absorption spectra exhibited blue shift and peak broadening; significant changes in 0–0 and 0–1 radiative transition intensity was found in photoluminescence emission spectra. The density of state (DOS) was directly mapped by energy resolved-electrochemical impedance spectroscopy (ER-EIS). Satellite states 0.5 eV below the lowest unoccupied molecular orbital (LUMO) band were revealed for the thinner polymer films. Moreover, the decreasing of the deep states density in the band gap manifested an increment in the material structural ordering with increasing thickness. Changes in the ratio between crystalline phases with face-on and edge-on orientation of F8BT chains were identified in the films by grazing-incidence wide angle X-ray scattering technique. A thickness threshold in all investigated aspects of the films at a thickness of about 100 nm was observed that can be attributed to the development of J-H aggregation in the film structure and mutual interplay between these two modes. Although a specific structure–property relationship thickness threshold value may be expected for thin films prepared from various polymers, solvents and under different process conditions, the value of about 100 nm can be generally considered as the characteristic length scale of this phenomenon.
Zn-containing Adhesives Facilitate Collagen Protection and Remineralization at the Resin-Dentin Interface: A Narrative Review
Authors: Toledano Manuel , Toledano-Osorio Manuel , Hannig Matthias , Carrasco-Carmona Álvaro , Osorio María T. , García-Godoy Franklin , Cabello Inmaculada , Osorio Raquel
Published: February 2022

This is a narrative review of the literature assessing the potential effectiveness of doping dentin polymeric adhesives with zinc compounds in order to improve bonding efficacy, remineralization and protection against degradation. A literature search was conducted using electronic databases, such as PubMed, MEDLINE, DIMDI and Web of Science. Through our search, we found literature demonstrating that Zn-doped dentin adhesives promote protection and remineralization of the resin-dentin interfaces. The increased bioactivity has also facilitated dentinal tubules’ occlusion by crystals’ precipitation contributing to improved sealing efficacy of restorations. Loading dentin adhesives with zinc gives rise to an increase of both crystallinity of mineral and crosslinking of collagen. The main role of zinc, in dentin adhesives, is to inhibit collagen proteolysis. We concluded that zinc exerts a protective effect through binding at the collagen-sensitive cleavage sites of matrix-metalloproteinases (MMPs), contributing to dentin matrix stabilization. Zinc may not only act as a MMPs inhibitor, but also influence signaling pathways and stimulate metabolic effects in dentin mineralization and remineralization processes. Zn-doped adhesives increase the longevity of dentin bonding through MMPs inhibition. Zn poses a remineralization strategy in demineralized dentin.
Integrating Taguchi Method and Gray Relational Analysis for Auto Locks by Using Multiobjective Design in Computer-Aided Engineering
Authors: Huang Wei-Tai , Tasi Zi-Yun , Ho Wen-Hsien , Chou Jyh-Horng
Published: February 2022

In automobiles, lock parts are matched with inserts, and this is a crucial quality standard for the dimensional accuracy of the molding. This study employed moldflow analysis to explore the influence of various injection molding process parameters on the warpage deformation. Deformation of the plastic part is caused by the nonuniform product temperature distribution in the manufacturing process. Furthermore, improper parameter design leads to substantial warpage and deformation. The Taguchi robust design method and gray correlation analysis were used to optimize the process parameters. Multiobjective quality analysis was performed for achieving a uniform temperature distribution and reducing the warpage deformation to obtain the optimal injection molding process parameters. Subsequently, three water cooling system designs—original cooling, U-shaped cooling, and conformal cooling—were tested to modify the temperature distribution and reduce the warpage. Taguchi gray correlation analysis revealed that the main influencing parameter was the mold temperature followed by the holding pressure. Moreover, the results indicated that the conformal cooling system improved the average temperature distribution.