bluuebunny/crossref_metadata_2025 huggingface dataset:
Survey on the Presence of Viruses of Economic and Zoonotic Importance in Avifauna in Northern Italy
Authors: Trogu Tiziana , Canziani Sabrina , Salvato Sara , Tolini Clara , Grilli Guido , Chiari Mario , Farioli Marco , Alborali Loris , Gaffuri Alessandra , Sala Giovanni , Bianchi Alessandro , Rosignoli Carlo , Prati Paola , Gradassi Matteo , Sozzi Enrica , Lelli Davide , Lavazza Antonio , Moreno Ana
Published: September 2021

Wild birds play an important role in the circulation and spread of pathogens that are potentially zoonotic or of high economic impact on zootechnical production. They include, for example, West Nile virus (WNV), Usutu virus (USUV), avian influenza virus (AIV), and Newcastle disease virus (NDV), which, despite having mostly an asymptomatic course in wild birds, have a strong impact on public health and zootechnical production. This study investigated the presence of these viruses in several wild bird species from North Italy during the biennium 2019–2020. Wild birds derived from 76 different species belonging to 20 orders. Out of 679 birds, 27 were positive for WNV (lineage 2) with a prevalence of 4%; all birds were negative for USUV; one gull was positive for H13N6 influenza virus, and 12 samples were positive for NDV with a prevalence of 2%. Despite the low prevalence observed, the analyses performed on these species provide further data, allowing a better understanding of the diffusion and evolution of diseases of both economic and zoonotic importance.
Changes in Antibiotic-Resistance Genes Induced by the Grazing Effect in Three Cladoceran Species
Authors: Choi Jong-Yun , Kim Seong-Ki
Published: September 2021

The acquisition of Antibiotic-Resistance Genes (ARGs) by natural bacteria caused by antibiotic abuse is causing serious problems for human and animal welfare. Here, we evaluated the influence of three cladoceran species on Antibiotic-Resistant Bacteria (ARB) and tetracycline-resistance gene (tet(A)) copies, and discussed the effect of these biological interactions on the distribution and diffusion of ARGs in freshwater ecosystems. Bacterial community and tet(A) abundances in water samples collected from wetlands were strongly influenced by cladoceran presence. The presence of Daphnia obtusa dramatically decreased ARB and tet(A) abundance compared to that with other cladoceran species (Chydorus sphaericus and Simocephalus vetulus). Interestingly, we found a high abundance of Flavobacteriales in the microbiomes of cladoceran species. Considering that Flavobacteriales species are potential carriers of the tet(A) gene, their adsorption and assimilation with cladocerans could significantly impact the reduction of tet(A) in water. Field surveys also showed that tet(A) abundance could be low if the dominance of D. obtusa in each wetland was high. This study highlighted the need for ecological interactions and a broad range of niches in the food web when discussing the fate of ARGs in freshwater ecosystems.
Promising Drug Targets and Compounds with Anti-Toxoplasma gondii Activity
Authors: Silva Marco da , Teixeira Cátia , Gomes Paula , Borges Margarida
Published: September 2021

Toxoplasmosis is a parasitic disease caused by the globally distributed protozoan parasite Toxoplasma gondii, which infects around one-third of the world population. This disease may result in serious complications for fetuses, newborns, and immunocompromised individuals. Current treatment options are old, limited, and possess toxic side effects. Long treatment durations are required since the current therapeutic system lacks efficiency against T. gondii tissue cysts, promoting the establishment of latent infection. This review highlights the most promising drug targets involved in anti-T. gondii drug discovery, including the mitochondrial electron transport chain, microneme secretion pathway, type II fatty acid synthesis, DNA synthesis and replication and, DNA expression as well as others. A description of some of the most promising compounds demonstrating antiparasitic activity, developed over the last decade through drug discovery and drug repurposing, is provided as a means of giving new perspectives for future research in this field.
Behaviour of Listeria monocytogenes and Natural Microflora during the Manufacture of Riojano Chorizo (Spanish Dry Cured Sausage)
Authors: Gonzalez-Fandos Elena , Vazquez de Castro Maria , Martinez-Laorden Alba
Published: September 2021

Riojano chorizo is a dry cured sausage manufactured with traditional technologies without adding starter cultures at low temperatures. Its characteristics differ from other types of chorizo since sugars and nitrites are no added and processing temperatures are low- This work evaluates the behaviour of Listeria monocytogenes during the processing of inoculated Riojano chorizo as well as the natural microflora that can play a technological role or be of interest as indicators. The sausage mixture was inoculated with a cocktail of three selected strains of L. monocytogenes (CECT 932, CECT 934 and CECT 4032) (4 log10 CFU/g) and after processed following the traditional production method. Samples were taken before inoculation, after inoculation, after stuffing (day 0) and on days 6, 13, 21 and 28 of processing. L. monocytogenes, mesophiles, Micrococcaceae, lactic acid bacteria, Enterobacteriaceae, S. aureus, sulfite-reducing clostridia and molds and yeast counts were evaluated. Furthermore, pH, water activity and humidity were determined. No growth of L mocytogenes was observed during the first 6 days, when the temperature of processing was 4 °C. The low temperature in the initial stages was a relevant hurdle to control L. monocytoegenes growth. A significant decrease (p ≤ 0.05) in L. monocytogenes counts was observed on day 13 compared to the initial counts. During drying (days 6 to 21) a reduction in this pathogen of 1.28 log CFU/g was observed. The low water activity below 0.92 on day 13 and 0.86 on day 21 seems to be critical for the reduction of L. monocytogenes.
Immunomodulatory Responses of Two Synthetic Peptides against Salmonella Typhimurium Infection
Authors: Ibarra-Valencia Marco Antonio , Espino-Solis Gerardo Pável , Estrada Blanca Elisa , Corzo Gerardo
Published: September 2021

In vitro assays of phagocytic activity showed that the peptide Pin2[G] stimulates phagocytosis in BMDM cells from 0.15 to 1.25 μg/mL, and in RAW 264.7 cells at 0.31 μg/mL. In the same way, the peptide FA1 induced phagocytosis in BMDM cells from 1.17 to 4.69 μg/mL and in RAW 264.7 cells at 150 μg/mL. Cytokine profiles of uninfected RAW 264.7 showed that Pin2[G] increased liberation TNF (from 1.25 to 10 μg/mL) and MCP-1 (10 μg/mL), and FA1 also increased the release of TNF (from 18.75 to 75 μg/mL) but did not increase the liberation of MCP-1. In RAW 264.7 macrophages infected with Salmonella enterica serovar Typhimurium, the expression of TNF increases with Pin2[G] (1.25–10 μg/mL) or FA1 (18.75–75 μg/mL). In these cells, FA1 also increases the expression of IL-12p70, IL-10 and IFN-γ when applied at concentrations of 37.5, 75 and 150 μg/mL, respectively. On the other hand, stimulation with 1.25 and 10 μg/mL of Pin2[G] promotes the expression of MCP-1 and IL-12p70, respectively. Finally, peptides treatment did not resolve murine gastric infection, but improves their physical condition. Cytokine profiles showed that FA1 reduces IFN-γ and MCP-1 but increases IL-10, while Pin2[G] reduces IFN-γ but increases the liberation of IL-6 and IL-12p70. This data suggests a promising activity of FA1 and Pin2[G] as immunomodulators of gastric infections in S. Typhimurium.
Soft Contact Lenses as Drug Delivery Systems: A Review
Authors: Rykowska Iwona , Nowak Iwona , Nowak Rafał
Published: September 2021

This review describes the role of contact lenses as an innovative drug delivery system in treating eye diseases. Current ophthalmic drug delivery systems are inadequate, particularly eye drops, which allow about 95% of the active substance to be lost through tear drainage. According to the literature, many interdisciplinary studies have been carried out on the ability of contact lenses to increase the penetration of topical therapeutic agents. Contact lenses limit drug loss by releasing the medicine into two layers of tears on either side of the contact lens, eventually extending the time of contact with the ocular surface. Thanks to weighted soft contact lenses, a continuous release of the drug over an extended period is possible. This article reviewed the various techniques to deliver medications through contact lenses, examining their advantages and disadvantages. In addition, the potential of drug delivery systems based on contact lenses has been extensively studied.
Regioselective Reduction of 1H-1,2,3-Triazole Diesters
Authors: Butler Christopher R. , Bendesky Justin , Schoffstall Allen Milton
Published: September 2021

Regioselective reactions can play pivotal roles in synthetic organic chemistry. The reduction of several 1-substituted 1,2,3-triazole 4,5-diesters by sodium borohydride has been found to be regioselective, with the C(5) ester groups being more reactive towards reduction than the C(4) ester groups. The amount of sodium borohydride and reaction time required for reduction varied greatly depending on the N(1)-substituent. The presence of a β-hydroxyl group on the N(1)-substituent was seen to have a rate enhancing effect on the reduction of the C(5) ester group. The regioselective reduction was attributed to the lower electron densities of the C(5) and the C(5) ester carbonyl carbon of the 1,2,3-triazole, which were further lowered in cases involving intramolecular hydrogen bonding.
Post-Operative Assessment of Ulnar Nerve Tension Using Shear-Wave Elastography
Authors: Durand Sebastien , Raffoul Wassim , Christen Thierry , Pedrazzi Nadine
Published: September 2021

Background: Ulnar nerve compression at the elbow level is the second-most common entrapment neuropathy. The aim of this study was to use shear-wave elastography for the quantification of ulnar nerve elasticity in patients after ulnar nerve decompression with anterior transposition and in the contralateral non-operative side. Method: Eleven patients with confirmed diagnosis and ulnar nerve decompression with anterior transposition were included and examinations were performed on an AixplorerTM ultrasound system (Supersonic Imagine, Aix-en-Provence, France). Results: We observed significant differences at 0-degree (p < 0.001), 45-degree (p < 0.05), 90-degree (p < 0.01) and 120-degree (p < 0.001) elbow flexion in the shear elastic modulus of the ulnar nerve in the operative and non-operative sides. There were no statistically significant differences between the elasticity values of the ulnar nerve after transposition at 0-degree elbow flexion and in the non-operative side at 120-degree elbow flexion (p = 0.39), or in the ulnar nerve after transposition at 120-degree elbow flexion and in the non-operative side at 0-degree elbow flexion (p = 0.09). Conclusion: Shear-wave elastography has the potential to be used postoperatively as a method for assessing nerve tension noninvasively by the estimation of mechanical properties, such as the shear elastic modulus.
Nitrogen Balance after the Administration of a Prolonged-Release Protein Substitute for Phenylketonuria as a Single Dose in Healthy Volunteers
Authors: Scheinin Mika , Junnila Jouni , Reiner Giorgio , MacDonald Anita , Muntau Ania C.
Published: September 2021

Nitrogen balance is the difference between nitrogen excreted as urea and nitrogen ingested, mainly in proteins. Increased circulating concentrations of amino acids (AA) in the bloodstream are usually associated with proportional increases in the production and excretion of urea. Previously, we reported results from a randomized, controlled, single-dose, crossover trial in healthy adult volunteers (n = 30) (Trial Registration: ISRCTN11016729), in which a Test product (prolonged-release AA mixture formulated with Physiomimic Technology™ (PT™)) significantly slowed down the release and reduced the peak plasma concentrations of essential AAs compared with a free AA mixture (Reference product) while maintaining essential AA bioavailability. Here, we report an assessment of the nitrogen balance from the same study. The amount of nitrogen contained in plasma AAs, levels of blood urea nitrogen (BUN) (p < 0.0001) and changes in BUN (p < 0.0001) were smaller after the Test product compared with the Reference product. These findings suggest that the production of urea in proportion to systemic AA availability was significantly smaller after the administration of the Test product compared with the Reference product and that the test product conferred the increased utilization of AAs for protein synthesis and reduced their oxidation and conversion to urea. In the clinical setting, it is possible that the effects of PT™ observed on the disposition of free AAs in this study may translate to health benefits in terms of physiological body composition and growth if used for the treatment of subjects with phenylketonuria (PKU). Further investigation in patients with PKU is warranted.
Human Milk Oligosaccharide 3′-GL Improves Influenza-Specific Vaccination Responsiveness and Immunity after Deoxynivalenol Exposure in Preclinical Models
Authors: Toutounchi Negisa Seyed , Braber Saskia , Hogenkamp Astrid , Varasteh Soheil , Cai Yang , Wehkamp Tjalling , Tims Sebastian , Leusink-Muis Thea , van Ark Ingrid , Wiertsema Selma , Stahl Bernd , Kraneveld Aletta D. , Garssen Johan , Folkerts Gert , van’t Land Belinda
Published: September 2021

Deoxynivalenol (DON), a highly prevalent mycotoxin food contaminant, is known to have immunotoxic effects. In the current study, the potential of dietary interventions with specific mixtures of trans-galactosyl-oligosaccharides (TOS) to alleviate these effects were assessed in a murine influenza vaccination model. Vaccine-specific immune responses were measured in C57Bl/6JOlaHsd mice fed diets containing DON, TOS or a combination, starting 2 weeks before the first vaccination. The direct effects of TOS and its main oligosaccharide, 3′-galactosyl-lactose (3′-GL), on DON-induced damage were studied in Caco-2 cells, as an in vitro model of the intestinal epithelial barrier. Exposure to DON significantly reduced vaccine-specific immune responses and the percentages of Tbet+ Th1 cells and B cells in the spleen. DON significantly altered epithelial structure and integrity in the ileum and reduced the SCFA levels in the cecum. Adding TOS into DON-containing diets significantly improved vaccine-specific immune responses, restored the immune cell balance in the spleen and increased SCFA concentrations in the cecum. Incubating Caco-2 cells with TOS and 3′-GL in vitro further confirmed their protective effects against DON-induced barrier disruption, supporting immune modulation. Overall, dietary intervention with TOS can attenuate the adverse effects of DON on Th1-mediated immune responses and gut homeostasis. These beneficial properties might be linked to the high levels of 3′-GL in TOS.
Association between Cardiovascular Health and Incident Atrial Fibrillation in the General Japanese Population Aged ≥40 Years
Authors: Nishikawa Tetsuo , Tanaka Yoshihiro , Tada Hayato , Tsuda Toyonobu , Kato Takeshi , Usui Soichiro , Sakata Kenji , Hayashi Kenshi , Kawashiri Masa-aki , Hashiba Atsushi , Takamura Masayuki
Published: September 2021

This study explores the association between lifestyle behavior and incident atrial fibrillation (AF) in the general Japanese population. Japanese residents aged ≥40 years undergoing a national health checkup in Kanazawa City were included. We hypothesized that better lifestyle behavior is associated with lower incidence of AF. Lifestyle behavior was evaluated by the total cardiovascular health (CVH) score (0 = poor to 14 = ideal), calculated as the sum of the individual scores on seven modifiable risk factors: smoking status, physical activity, obesity, patterns of eating schedule, blood pressure, total cholesterol, and blood glucose. The association between CVH and incident AF was assessed, adjusting for other factors. A total of 37,523 participants (mean age 72.3 ± 9.6 years, 36.8% men, and mean total CVH score 9 ± 1) were analyzed. During the median follow-up period of 5 years, 703 cases of incident AF were observed. Using a low CVH score as a reference, the upper group (ideal CVH group) had a significantly lower risk of incident AF (hazard ratio [HR] = 0.79, 95% confidence interval 0.65–0.96, p = 0.02), especially among those aged <75 years (HR = 0.68, 95% confidence interval 0.49–0.94, p = 0.02). Thus, ideal CVH is independently associated with a lower risk for incident AF, particularly in younger Japanese individuals (<75 years).
Being Underweight Increases the Risk of Non-Cystic Fibrosis Bronchiectasis in the Young Population: A Nationwide Population-Based Study
Authors: Yang Bumhee , Han Kyungdo , Kim Sang Hyuk , Lee Dong-Hwa , Park Sang Hyun , Yoo Jung Eun , Shin Dong Wook , Choi Hayoung , Lee Hyun
Published: September 2021

Although body mass index (BMI) is a potential risk factor for bronchiectasis in young adults, the association between BMI and incident bronchiectasis has not been well elucidated. This study included 6,329,838 individuals aged 20–40 years from the Korean National Health Insurance Service database 2009–2012 who were followed up until the date of the diagnosis of bronchiectasis, death, or 31 December 2018. We evaluated the incidence and risk of bronchiectasis according to the BMI category. The incidence rate of bronchiectasis increased as BMI decreased in a dose-dependent manner (p for trend <0.01). In multivariable Cox regression analysis, being underweight was an independent risk factor for the development of bronchiectasis, with a hazard ratio of 1.24 (95% confidence interval, 1.19–1.30) compared to being normal weight. In subgroup analysis, the effect of being underweight on the development of bronchiectasis was more evident in males and older individuals (30–40 years) than females and younger individuals (20–29 years), respectively (p for interaction <0.01 for both). These results remained significant in subgroup analysis in which subjects with comorbidities related to being underweight were excluded. Being underweight may be a novel risk factor for the development of bronchiectasis in young adults.
Association Study among Comethylation Modules, Genetic Polymorphisms and Clinical Features in Mexican Teenagers with Eating Disorders: Preliminary Results
Authors: Nolasco-Rosales Germán Alberto , Martínez-Magaña José Jaime , Juárez-Rojop Isela Esther , González-Castro Thelma Beatriz , Tovilla-Zarate Carlos Alfonso , García Ana Rosa , Sarmiento Emmanuel , Ruiz-Ramos David , Genis-Mendoza Alma Delia , Nicolini Humberto
Published: September 2021

Eating disorders are psychiatric disorders characterized by disturbed eating behaviors. They have a complex etiology in which genetic and environmental factors interact. Analyzing gene-environment interactions could help us to identify the mechanisms involved in the etiology of such conditions. For example, comethylation module analysis could detect the small effects of epigenetic interactions, reflecting the influence of environmental factors. We used MethylationEPIC and Psycharray microarrays to determine DNA methylation levels and genotype from 63 teenagers with eating disorders. We identified 11 comethylation modules in WGCNA (Weighted Gene Correlation Network Analysis) and correlated them with single nucleotide polymorphisms (SNP) and clinical features in our subjects. Two comethylation modules correlated with clinical features (BMI and height) in our sample and with SNPs associated with these phenotypes. One of these comethylation modules (yellow) correlated with BMI and rs10494217 polymorphism (associated with waist-hip ratio). Another module (black) was correlated with height, rs9349206, rs11761528, and rs17726787 SNPs; these polymorphisms were associated with height in previous GWAS. Our data suggest that genetic variations could alter epigenetics, and that these perturbations could be reflected as variations in clinical features.
Electrocyclizations of Conjugated Azapolyenes Produced in Reactions of Azaheterocycles with Metal Carbenes
Authors: Rostovskii Nikolai V. , Novikov Mikhail S. , Khlebnikov Alexander F.
Published: September 2021

Conjugated azapolyenes (azabuta-1,3-dienes, aza-/diaza-/oxaza-/oxadiazahexa-1,3,5-trienes) are highly reactive in electrocyclization reactions, which makes them convenient precursors for the synthesis of a wide range of four-, five-, and six-membered nitrogen heterocycles that are of relevance for medicinal chemistry. Ring opening reactions of 2H-azirines and azoles containing an N–N or N–O bond, initiated by a transition metal carbene, have become increasingly important in recent years, since they easily allow the generation of azapolyenes with different numbers of double bonds and heteroatoms in various positions. This review summarizes the literature, published mainly in the last decade, on the synthetic and mechanistic aspects of electrocyclizations of azapolyenes generated by the carbene method.
Detection of Anaplasma phagocytophilum in Wild and Farmed Cervids in Poland
Authors: Myczka Anna W. , Steiner-Bogdaszewska Żaneta , Filip-Hutsch Katarzyna , Oloś Grzegorz , Czopowicz Michał , Laskowski Zdzisław
Published: September 2021

Background: The role of cervids in the circulation of A. phagocytophilum has not yet been clearly determined; however, several species of wild and farm cervids may be a natural reservoir of this bacteria. Methods: Spleen and liver tissue samples were taken from 207 wild (red deer, roe deer, fallow deer and moose) and farmed cervids (red deer and fallow deer) from five geographical areas. These were tested for the A. phagocytophilum16S rDNA partial gene by nested PCR. Results: Anaplasma spp. were detected in 91 of 207 examined cervids (prevalence 43.9%). Three different variants of 16S rDNA partial gene were reported, one for the first time. Anaplasma phagocytophilum was more often detected in young specimens than in adults and more often in the spleen than in the liver. Conclusions: Cervids from the four sites across Poland were found to be major natural reservoirs of various strains of A. phagocytophilum. This is the first study to use spleen and liver as biological material to detect A. phagocytophilum in moose in Poland.
Proteomic Analysis of Leishmania donovani Membrane Components Reveals the Role of Activated Protein C Kinase in Host-Parasite Interaction
Authors: Verma Sandeep , Deep Deepak Kumar , Gautam Poonam , Singh Ruchi , Salotra Poonam
Published: September 2021

Visceral leishmaniasis (VL), mainly caused by the Leishmania donovani parasitic infection, constitutes a potentially fatal disease, for which treatment is primarily dependent on chemotherapy. The emergence of a resistant parasite towards current antileishmanial agents and increasing reports of relapses are the major concerns. Detailed research on the molecular interaction at the host-parasite interface may provide the identification of the parasite and the host-related factors operating during disease development. Genomic and proteomic studies highlighted several essential secretory and cytosolic proteins that play vital roles during Leishmania pathogenesis. The aim of this study was to identify membrane proteins from the Leishmania donovani parasite and the host macrophage that interact with each other using 2-DE/MALDI-TOF/MS. We identified membrane proteins including activated protein C kinase, peroxidoxin, small myristoylated protein 1 (SMP-1), and cytochrome C oxidase from the parasite, while identifying filamin A interacting protein 1(FILIP1) and β-actin from macrophages. We further investigated parasite replication and persistence within macrophages following the macrophage-amastigote model in the presence or absence of withaferin (WA), an inhibitor of activated C kinase. WA significantly reduced Leishmania donovani replication within host macrophages. This study sheds light on the important interacting proteins for parasite proliferation and virulence, and the establishment of infection within host cells, which can be targeted further to develop a strategy for chemotherapeutic intervention.
Immunobiotic Lactobacilli Improve Resistance of Respiratory Epithelial Cells to SARS-CoV-2 Infection
Authors: Islam Md. Aminul , Albarracin Leonardo , Tomokiyo Mikado , Valdez Juan Carlos , Sacur Jacinto , Vizoso-Pinto Maria Guadalupe , Andrade Bruno G. N. , Cuadrat Rafael R. C. , Kitazawa Haruki , Villena Julio
Published: September 2021

Previously, we reported that immunomodulatory lactobacilli, nasally administered, beneficially regulated the lung antiviral innate immune response induced by Toll-like receptor 3 (TLR3) activation and improved protection against the respiratory pathogens, influenza virus and respiratory syncytial virus in mice. Here, we assessed the immunomodulatory effects of viable and non-viable Lactiplantibacillus plantarum strains in human respiratory epithelial cells (Calu-3 cells) and the capacity of these immunobiotic lactobacilli to reduce their susceptibility to the acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Immunobiotic L. plantarum MPL16 and CRL1506 differentially modulated IFN-β, IL-6, CXCL8, CCL5 and CXCL10 production and IFNAR2, DDX58, Mx1 and OAS1 expression in Calu-3 cells stimulated with the TLR3 agonist poly(I:C). Furthermore, the MPL16 and CRL1506 strains increased the resistance of Calu-3 cells to the challenge with SARS-CoV-2. L. plantarum MPL16 induced these beneficial effects more efficiently than the CRL1506 strain. Of note, neither non-viable MPL16 and CRL1506 strains nor the non-immunomodulatory strains L. plantarum CRL1905 and MPL18 could modify the resistance of Calu-3 cells to SARS-CoV-2 infection or the immune response to poly(I:C) challenge. To date, the potential beneficial effects of immunomodulatory probiotics on SARS-CoV-2 infection and COVID-19 outcome have been extrapolated from studies carried out in the context of other viral pathogens. To the best of our knowledge, this is the first demonstration of the ability of immunomodulatory lactobacilli to positively influence the replication of the new coronavirus. Further mechanistic studies and in vivo experiments in animal models of SARS-CoV-2 infection are necessary to identify specific strains of beneficial immunobiotic lactobacilli like L. plantarum MPL16 or CRL1506 for the prevention or treatment of the COVID-19.
New Prospects in Neutering Male Animals Using Magnetic Nanoparticle Hyperthermia
Authors: Jivago José Luiz P. R. , Brito Juliana Lis Mendes , Capistrano Gustavo , Vinícius-Araújo Marcus , Lima Verde Ediron , Bakuzis Andris Figueiroa , Souza Paulo E. N. , Azevedo Ricardo Bentes , Lucci Carolina Madeira
Published: September 2021

Controlling populations of free-roaming dogs and cats poses a huge challenge worldwide. Non-surgical neutering strategies for male animals have been long pursued, but the implementation of the procedures developed has remained limited to date. As submitting the testes to high temperatures impairs spermatogenesis, the present study investigated localized application of magnetic nanoparticle hyperthermia (MNH) to the testicles as a potential non-surgical sterilization method for animals. An intratesticular injection of a magnetic fluid composed of manganese-ferrite nanoparticles functionalized with citrate was administered followed by testicle exposure to an alternate magnetic field to generate localized heat. Testicular MNH was highly effective, causing progressive seminiferous tubule degeneration followed by substitution of the parenchyma with stromal tissue and gonadal atrophy, suggesting an irreversible process with few side effects to general animal health.
Combining Photodynamic Therapy with Immunostimulatory Nanoparticles Elicits Effective Anti-Tumor Immune Responses in Preclinical Murine Models
Authors: Huis in ‘t Veld Ruben Victor , Da Silva Candido G. , Jager Martine J. , Cruz Luis J. , Ossendorp Ferry
Published: September 2021

Photodynamic therapy (PDT) has shown encouraging but limited clinical efficacy when used as a standalone treatment against solid tumors. Conversely, a limitation for immunotherapeutic efficacy is related to the immunosuppressive state observed in large, advanced tumors. In the present study, we employ a strategy, in which we use a combination of PDT and immunostimulatory nanoparticles (NPs), consisting of poly(lactic-co-glycolic) acid (PLGA)-polyethylene glycol (PEG) particles, loaded with the Toll-like receptor 3 (TLR3) agonist poly(I:C), the TLR7/8 agonist R848, the lymphocyte-attracting chemokine, and macrophage inflammatory protein 3α (MIP3α). The combination provoked strong anti-tumor responses, including an abscopal effects, in three clinically relevant murine models of cancer: MC38 (colorectal), CT26 (colorectal), and TC-1 (human papillomavirus 16-induced). We show that the local and distal anti-tumor effects depended on the presence of CD8+ T cells. The combination elicited tumor-specific oncoviral- or neoepitope-directed CD8+ T cells immune responses against the respective tumors, providing evidence that PDT can be used as an in situ vaccination strategy against cancer (neo)epitopes. Finally, we show that the treatment alters the tumor microenvironment in tumor-bearing mice, from cold (immunosuppressed) to hot (pro-inflammatory), based on greater neutrophil infiltration and higher levels of inflammatory myeloid and CD8+ T cells, compared to untreated mice. Together, our results provide a rationale for combining PDT with immunostimulatory NPs for the treatment of solid tumors.
Assessment of the Effect of A-PRF Application during the Surgical Extraction of Third Molars on Healing and the Concentration of C-Reactive Protein
Authors: Nowak Jacek M. , Surma Stanisław , Romańczyk Monika , Wojtowicz Andrzej , Filipiak Krzysztof J. , Czerniuk Maciej R.
Published: September 2021

Extraction procedures for mandibular third molars are performed all over the world every day. Local inflammation resulting from surgery, and the pain that patients experience, often make it impossible to take up daily life activities, such as work or sports. Growth and anti-inflammatory factors, located in the fibrin network, have a positive effect on tissue-healing processes and should also reduce local inflammation. Advanced platelet-rich fibrin (A-PRF) applied locally influences such processes as: angiogenesis, osteogenesis and collagenogenesis. It also affects mesenchymal cell lines and anti- and pro-inflammatory mediators. Due to the autologous origin of the material, their use in guide bone regeneration (GBR) is more and more widespread in dentistry. The results of previous studies indicate that the use of A-PRF in the treatment area significantly reduces postoperative pain, while the formation of edema is not affected. C-reactive protein (CRP), which is an acute phase protein, appears in the blood as a consequence of inflammation. Due to the dynamics of changes in concentration of CRP, it is a protein that is sufficiently sensitive and is used in studies to monitor the tissue healing process. The effect of A-PRF application on CRP concentrations, before and after surgery, has not been investigated yet. The study was conducted on 60 generally healthy patients. A faster decrease of CRP levels was shown in patients who used A-PRF after the procedure. Additionally, it accelerated healing and reduced the occurrence of a dry socket close to 0.
Effect of Salicylic Acid in the Yield of Ricinine in Ricinus communis under Greenhouse Condition
Authors: Zavala-Gómez Carlos Eduardo , Rodríguez-deLeón Eloy , Bah Mamadou Moustapha , Feregrino-Pérez Ana Angélica , Campos-Guillén Juan , Amaro-Reyes Aldo , Rodríguez-Morales José Alberto , García-Trejo Juan Fernando , Flores-Macias Antonio , Figueroa-Brito Rodolfo , Ramos-López Miguel Angel
Published: September 2021

Castor bean (Ricinus communis) seeds contain ricinine, an alkaloid with insecticidal and insectistatic activities. Elicitation with salicylic acid (SA) has proven to stress R. communis and might modify the ricinine concentration. The aim of this study was to evaluate the concentration of ricinine in the bagasse of seeds from R. communis elicited with exogenous SA under greenhouse conditions. Plants were grown and divided into five groups, which were sprayed with SA and drench with 50 mL 60 days after sowing with concentrations of SA (0, 100, 300, 600 and 900 µM). Clusters were mixed and separated according to the treatment, and dried. The seeds were ground, the oil was extracted by Soxhlet with hexane, and then the bagasse was extracted with methanol. Ricinine was determined by HPLC. Elicitation did not change the plant height or diameter; the control group had 9.17 µg mL−1 of ricinine; and the concentrations followed a hormesis curve with the peak at 300 µM of SA that had a ricinine concentration of 18.25 µg mL−1. Elicitation with SA might be a cost-effective technique to increase ricinine from R. communis bagasse.
Genome-Wide Identification and Characterization of the Abiotic-Stress-Responsive GRF Gene Family in Diploid Woodland Strawberry (Fragaria vesca)
Authors: Li Zhiqi , Xie Qian , Yan Jiahui , Chen Jianqing , Chen Qingxi
Published: September 2021

Growth regulatory factors (GRF) are plant-specific transcription factors that play an important role in plant resistance to stress. This gene family in strawberry has not been investigated previously. In this study, 10 GRF genes were identified in the genome of the diploid woodland strawberry (Fragaria vesca). Chromosome analysis showed that the 10 FvGRF genes were unevenly distributed on five chromosomes. Phylogenetic analysis resolved the FvGRF proteins into five groups. Genes of similar structure were placed in the same group, which was indicative of functional redundance. Whole-genome duplication/segmental duplication and dispersed duplication events effectively promoted expansion of the strawberry GRF gene family. Quantitative reverse transcription-PCR analysis suggested that FvGRF genes played potential roles in the growth and development of vegetative organs. Expression profile analysis revealed that FvGRF3, FvGRF5, and FvGRF7 were up-regulated under low-temperature stress, FvGRF4 and FvGRF9 were up-regulated under high-temperature stress, FvGRF6 and FvGRF8 were up-regulated under drought stress, FvGRF3, FvGRF6, and FvGRF8 were up-regulated under salt stress, FvGRF2, FvGRF7, and FvGRF9 were up-regulated under salicylic acid treatment, and FvGRF3, FvGRF7, FvGRF9, and FvGRF10 were up-regulated under abscisic acid treatment. Promoter analysis indicated that FvGRF genes were involved in plant growth and development and stress response. These results provide a theoretical and empirical foundation for the elucidation of the mechanisms of abiotic stress responses in strawberry.
A Novel and Efficient In Vitro Organogenesis Approach for Ajuga lupulina Maxim
Authors: Wu Qinggui , Yang Honglin , Yang Yulin , He Jinyu , Aer Erga , Ma Yonghong , Zou Lijuan
Published: September 2021

This work was aimed at establishing an effective approach for in vitro propagation of Ajuga lupulina Maxim, a medicinal and ornamental plant mainly found in eastern Xizang, in the western Sichuan region of China. We report an optimum response in the proliferation of axillary shoots from nodal segment explants (10.2 shoots/explant) on MS medium containing 3.0 mg L−1 of 6-benzyladenine (BA). When BA and TDZ individually or in combination with NAA were employed for adventitious shoot regeneration, shoots and embryo-like structures (ELSs) were noted in the callus from leaf explants. The maximum response of 26.4 shoots /explant (81.6%) and 12.0 ELSs/explant were ascertained on MS medium with 4.0 mg L−1 TDZ and 0.1 mg L−1 NAA. The leaf despite browning still demonstrated a high regeneration capacity. TDZ (2.0 mg L−1) and BA (2.0 mg L−1) along with NAA (0.01 mg L−1) were found to perform well for shoot regeneration via callus from shoot tip explants. The best for rooting was MS medium (half-strength) containing indole-3-butyric acid (IBA: 1.5 mg L−1) and (NAA: 0.5 mg L−1) with the maximum number of roots (25.8 per shoot) and the highest rooting frequency (81.71%). The survival of the plantlets in the greenhouse was 78.2% indicative of successful acclimatization. This work is the first report of a consistent, definitive, and unique protocol for A. lupulina regeneration, paving the way for the in vitro preservation of such significant genetic resources and also further allied systems based on such callus-based or embryo-based approaches.
UHPLC-Orbitrap-MS Tentative Identification of 51 Oleraceins (Cyclo-Dopa Amides) in Portulaca oleracea L. Cluster Analysis and MS2 Filtering by Mass Difference
Authors: Voynikov Yulian , Nedialkov Paraskev , Gevrenova Reneta , Zheleva-Dimitrova Dimitrina , Balabanova Vessela , Dimitrov Ivan
Published: September 2021

Oleraceins are a class of indoline amide glycosides found in Portulaca oleracea L. (Portulacaceae), or purslane. These compounds are characterized by 5,6-dihydroxyindoline-2-carboxylic acid N-acylated with cinnamic acid derivatives, and many are glucosylated. Herein, hydromethanolic extracts of the aerial parts of purslane were subjected to UHPLC-Orbitrap-MS analysis, in negative ionization mode. Diagnostic ion filtering (DIF), followed by diagnostic difference filtering (DDF), were utilized to automatically filter out MS data and select plausible oleracein structures. After an in-depth MS2 analysis, a total of 51 oleracein compounds were tentatively identified. Of them, 26 had structures, matching one of the already known oleracein, and the other 25 were new, undescribed in the literature compounds, belonging to the oleracein class. Moreover, based on selected diagnostic fragment ions, clustering algorithms and visualizations were utilized. As we demonstrate, clustering methods provide valuable insights into the mass fragmentation elucidation of natural compounds in complex mixtures.
Influence of Silica Specific Surface Area on the Viscoelastic and Fatigue Behaviors of Silica-Filled SBR Composites
Authors: Padmanathan Hiron Raja , Federico Carlos Eloy , Addiego Frédéric , Rommel Robert , Kotecký Ondřej , Westermann Stephan , Fleming Yves
Published: September 2021

This work aimed at studying the effect of a silica specific surface area (SSA), as determined by the nitrogen adsorption method, on the viscoelastic and fatigue behaviors of silica-filled styrene–butadiene rubber (SBR) composites. In particular, silica fillers with an SSA of 125 m2/g, 165 m2/g, and 200 m2/g were selected. Micro-computed X-ray tomography (µCT) was utilized to analyze the 3D morphology of the fillers within an SBR matrix prior to mechanical testing. It was found with this technique that the volume density of the agglomerates drastically decreased with decreasing silica SSA, indicating an increase in the silica dispersion state. The viscoelastic behavior was evaluated by dynamic mechanical analysis (DMA) and hysteresis loss experiments. The fatigue behavior was studied by cyclic tensile loading until rupture enabled the generation of Wöhler curves. Digital image correlation (DIC) was used to evaluate the volume strain upon deformation, whereas µCT was used to evaluate the volume fraction of the fatigue-induced cracks. Last, scanning electron microscopy (SEM) was used to characterize, in detail, crack mechanisms. The main results indicate that fatigue life increased with decreasing silica SSA, which was also accompanied by a decrease in hysteresis loss and storage modulus. SEM investigations showed that filler–matrix debonding and filler fracture were the mechanisms at the origin of crack initiation. Both the volume fraction of the cracks obtained by µCT and the volume strain acquired from the DIC increased with increasing SSA of silica. The results are discussed based on the prominent role of the filler network on the viscoelastic and fatigue damage behaviors of SBR composites.
A Hybrid Cooling Model Based on the Use of Newly Designed Fluted Conformal Cooling Channels and Fastcool Inserts for Green Molds
Authors: Torres-Alba Abelardo , Mercado-Colmenero Jorge Manuel , Caballero-Garcia Juan De Dios , Martin-Doñate Cristina
Published: September 2021

The paper presents a hybrid cooling model based on the use of newly designed fluted conformal cooling channels in combination with inserts manufactured with Fastcool material. The hybrid cooling design was applied to an industrial part with complex geometry, high rates of thickness, and deep internal concavities. The geometry of the industrial part, besides the ejection system requirements of the mold, makes it impossible to cool it adequately using traditional or conformal standard methods. The addition of helical flutes in the circular conformal cooling channel surfaces generates a high number of vortexes and turbulences in the coolant flow, fostering the thermal exchange between the flow and the plastic part. The use of a Fastcool insert allows an optimal transfer of the heat flow in the slender core of the plastic part. An additional conformal cooling channel layout was required, not for the cooling of the plastic part, but for cooling the Fastcool insert, improving the thermal exchange between the Fastcool insert and the coolant flow. In this way, it is possible to maintain a constant heat exchange throughout the manufacturing cycle of the plastic part. A transient numerical analysis validated the improvements of the hybrid design presented, obtaining reductions in cycle time for the analyzed part by 27.442% in comparison with traditional cooling systems. The design of the 1 mm helical fluted conformal cooling channels and the use of the Fastcool insert cooled by a conformal cooling channel improves by 4334.9% the thermal exchange between the cooling elements and the plastic part. Additionally, it improves by 51.666% the uniformity and the gradient of the temperature map in comparison with the traditional cooling solution. The results obtained in this paper are in line with the sustainability criteria of green molds, centered on reducing the cycle time and improving the quality of the complex molded parts.
The Surface Characterisation of Fused Filament Fabricated (FFF) 3D Printed PEEK/Hydroxyapatite Composites
Authors: Rodzeń Krzysztof , McIvor Mary Josephine , Sharma Preetam K. , Acheson Jonathan G. , McIlhagger Alistair , Mokhtari Mozaffar , McFerran Aoife , Ward Joanna , Meenan Brian J. , Boyd Adrian R.
Published: September 2021

Polyetheretherketone (PEEK) is a high-performance thermoplastic polymer which has found increasing application in orthopaedics and has shown a lot of promise for ‘made-to-measure’ implants via additive manufacturing approaches. However, PEEK is bioinert and needs to undergo surface modification to make it at least osteoconductive to ensure a more rapid, improved, and stable fixation that will last longer in vivo. One approach to solving this issue is to modify PEEK with bioactive agents such as hydroxyapatite (HA). The work reported in this study demonstrates the direct 3D printing of PEEK/HA composites of up to 30 weight percent (wt%) HA using a Fused Filament Fabrication (FFF) approach. The surface characteristics and in vitro properties of the composite materials were investigated. X-ray diffraction revealed the samples to be semi-crystalline in nature, with X-ray Photoelectron Spectroscopy and Time-of-Flight Secondary Ion Mass Spectrometry revealing HA materials were available in the uppermost surface of all the 3D printed samples. In vitro testing of the samples at 7 days demonstrated that the PEEK/HA composite surfaces supported the adherence and growth of viable U-2 OS osteoblast like cells. These results demonstrate that FFF can deliver bioactive HA on the surface of PEEK bio-composites in a one-step 3D printing process.
Removal of Nitrate Nitrogen by Rhodotorula graminis Immobilized in Alginate Gel for Groundwater Treatment
Authors: Nishida Masashi , Matsuo Syuhei , Yamanari Karin , Iwahara Masayoshi , Kusakabe Katsuki
Published: September 2021

Groundwater is the source of all tap water in Kumamoto City, Japan. However, the concentration of nitrate nitrogen (NO3−-N) tends to increase every year due to the influences of overfertilization, field disposal of livestock manure, and inflow of domestic wastewater. A heterotrophic nitrification–aerobic denitrification (HN-AD) system is an attractive approach for nitrate-nitrogen removal. In this study, Rhodotorula graminis NBRC0190, a naturally occurring red yeast that shows high nitrogen removal performance in glucose, was immobilized on calcium alginate hydrogel beads. NO3−-N removal efficiency exceeded 98% in the region of NO3−-N concentration below 10 mg/L in the model groundwater. Even after the same treatment was repeated five times, the denitrification performance of the R. gra immobilized alginate hydrogel beads was maintained. Finally, when this treatment method was applied to actual groundwater in Kumamoto City, it was possible to make the water of even higher quality.
Reduction of Volatile Organic Compounds (VOCs) Emissions from Laundry Dry-Cleaning by an Integrated Treatment Process of Condensation and Adsorption
Authors: Song Mugeun , Kim Kyunghoon , Cho Changmin , Kim Daekeun
Published: September 2021

Volatile organic compounds (VOCs) are intermittently emitted at high concentrations (tens of thousands of ppmv) from small-scale laundry shops in urban areas, affecting the urban atmospheric environment. In this study, we suggested integrating VOC treatment processes incorporating condensation and adsorption in series to remove VOCs released from small-scale laundry dryers (laundry weighing less than 30 kg). We designed two different processes depending on regeneration modes for adsorber beds; an open-circuit flow process and a closed-loop flow process in regeneration mode. Our VOC treatment processes enable sustainable operation via the regeneration of adsorbers on a regular basis. Before applying the VOC treatment processes, average concentration of total volatile organic compounds (TVOCs) was 4099 ppmv (12,000 ppmv of the peak concentration) during the drying operation. After applying our closed-loop flow process, TVOC concentration decreased to 58 ppmv, leading to 98.5% removal efficiency. We also verified the robustness of our process performance in a continuous operation (30 cycles) by using a process simulation program. Lastly, we observed that our integrated treatment process can contribute to reductions in ozone and secondary organic aerosol generation by 90.4% and 95.9%, respectively. We concluded that our integrated VOC treatment processes are applicable to small-scale laundry shops releasing high-concentration VOCs intermittently, and are beneficial to the atmospheric environment.
An Intelligent Process Fault Diagnosis System Based on Neural Networks and Andrews Plot
Authors: Wang Shengkai , Zhang Jie
Published: September 2021

This paper proposes a neural network-based process fault diagnosis system with Andrews plot for information pre-processing to enhance the performance of online process fault diagnosis. By using features extracted from Andrews plot as the inputs to a neural network, as a classifier, the diagnosis speed and reliability are improved. A method for determining the important features in the Andrews function is proposed. The proposed fault diagnosis system is applied to a simulated continuous stirred tank reactor process and is compared with two conventional neural network-based fault diagnosis systems: scheme B where the monitored measurements are directly fed to a neural network after scaling and scheme C where the monitored measurements are converted to qualitative trend data before feeding to a neural network. Of all the considered faults, the proposed fault diagnosis system diagnosed the abrupt faults on average 5.45 s and 2.66 s earlier than schemes B and C, respectively and diagnosed the incipient faults on average 13.82 s and 5.09 s earlier than schemes B and C, respectively. The results reveal that Andrews plot method utilized in online process monitoring has a great potential in industrial process monitoring.