bluuebunny/crossref_metadata_2025 huggingface dataset:
An Electrospun Scaffold Loaded with an Enteromorpha Polysaccharide for Accelerated Wound Healing in Diabetic Mice
Authors: Guo Lili , Guan Na , Miao Wenjun , Zhao Wenwen , Li Qiu
Published: January 2022

The design and development of innovative multifunctional wound dressing materials in engineered biomaterials is essential for promoting tissue repair. In this study, nanofibrous wound dressing materials loaded with anti-inflammatory ingredients were manufactured by a promising electrospinning strategy, and their capability for treating diabetic wounds was also investigated. A scaffold blend consisting of an Enteromorpha polysaccharide and polyvinyl alcohol (PVA) was fabricated. The in vitro and in vivo studies confirmed the efficacy of PVA/EPP1 fiber. We found that PVA/EPP1 fiber accelerated the repair of a full-thickness skin wound in diabetic mice. The results suggest that this scaffold could effectively shorten the wound healing time by inhibiting inflammatory activity, which makes it a promising candidate for the treatment of hard-to-heal wounds caused by diabetes.
Novel Antioxidant Peptides from Crassostrea Hongkongensis Improve Photo-Oxidation in UV-Induced HaCaT Cells
Authors: Zhang Chen , Lv Jiatong , Qin Xiaoming , Peng Zhilan , Lin Haisheng
Published: January 2022

Enzymatic hydrolysates from Oysters (OAH) display multiple biological activities. Previously, a 3~5 KDa oyster ultrafiltration component (OUP) showed a high property of preventing skin oxidation. Subsequently, we identified specific peptides with such activity. OUP was fractionated stepwise by Sephadex-G25 and RP-HPLC, and active fractions were screened using UV-irradiated HaCaT cells. The most active fractions (OP5-3) were analyzed by LC-MS/MS and a total of 17 peptides were identified. Results from mass spectrometry showed that OP5-3 consisted of peptides with a molecular weight range of 841.51–1786.92 Da. Six of these peptides were synthesized for validating the activity of resisting skin oxidation in the same cell model. All six peptides showed varying degrees of antioxidant activity, while pretreatment of HaCaT cells with AIVAEVNEAAK alleviated UV cytotoxicity, inhibited metalloproteinase 1 (MMP-1) expression, and showed the highest activity to resist UV-induced skin photo-oxidation among these peptides. In addition, results from molecular docking analysis of MMP-1 with AIVAEVNEAAK showed that AIVAEVNEAAK suppresses its enzymatic activity by directly interacting with MMP-1 and thus exhibit anti-photoaging activity.
Chitosan-Based Films with 2-Aminothiophene Derivative: Formulation, Characterization and Potential Antifungal Activity
Authors: Oliveira Verônica da Silva , Cruz Meriângela Miranda da , Bezerra Gabriela Suassuna , Silva Natan Emanuell Sobral e , Nogueira Fernando Henrique Andrade , Chaves Guilherme Maranhão , Sobrinho José Lamartine Soares , Mendonça-Junior Francisco Jaime Bezerra , Damasceno Bolívar Ponciano Goulart de Lima , Converti Attilio , Lima Ádley Antonini Neves de
Published: January 2022

In this study, films of chitosan and 2-amino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carbonitrile (6CN), a 2-aminothiophene derivative with great pharmacological potential, were prepared as a system for a topical formulation. 6CN-chitosan films were characterized by physicochemical analyses, such as Fourier-transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD), and scanning electronic microscopy (SEM). Additionally, the antifungal potential of the films was evaluated in vitro against three species of Candida (C. albicans, C. tropicalis, and C. parapsilosis). The results of the FTIR and thermal analysis showed the incorporation of 6CN in the polymer matrix. In the diffractogram, the 6CN-chitosan films exhibited diffraction halos that were characteristic of amorphous structures, while the micrographs showed that 6CN particles were dispersed in the chitosan matrix, exhibiting pores and cracks on the film surface. In addition, the results of antifungal investigation demonstrated that 6CN-chitosan films were effective against Candida species showing potential for application as a new antifungal drug.
Relation between Handgrip Strength and Quality of Life in Patients with Arthritis in Korea: The Korea National Health and Nutrition Examination Survey, 2015–2018
Authors: Chang So-Youn , Han Byoung-Duck , Han Kyung-Do , Park Hyo-Jin , Kang Seok
Published: January 2022

Background and Objectives: This study aimed to investigate the relationship between handgrip strength (HGS) and quality of life (QOL) in patients diagnosed with osteoarthritis (OA) or rheumatoid arthritis (RA). Materials and Methods: We enrolled 13,966 from the Korea National Health and Nutrition Examination Survey from 2015 to 2018. All participants underwent the health-related QOL assessment using the European Quality of Life Scale-Five dimensions (EQ-5D) and measured the HGS. The weak HGS was defined as the lowest quartile. We investigated the difference in QOL between patients with arthritis and the healthy control group and evaluated the correlation between weak HGS and QOL in arthritis patients. Results: Those diagnosed with OA or RA had significantly lower QOL than healthy controls. The weak HGS was significantly correlated with lower QOL in arthritis patients. Among OA patients, those with weak HGS revealed significantly higher odds ratios for impairment in all dimensions of EQ-5D. RA patients with weak HGS had significantly higher odds ratios for impairment in dimensions of mobility, self-care, usual activity, and pain/discomfort than those with normal HGS. Conclusions: These results suggest that weak HGS is significantly associated with decreased QOL in patients with arthritis.
Effects of Different Drug Therapies and COVID-19 mRNA Vaccination on Semen Quality in a Man with Ankylosing Spondylitis: A Case Report
Authors: Chatzimeletiou Katerina , Fleva Alexandra , Sioga Antonia , Georgiou Ioannis , Nikolopoulos Theodoros-Thomas , Markopoulou Maria , Petrogiannis Nikos , Anifandis George , Patrikiou Antonios , Kolibianakis Efstratios , Giannakou Anastasia , Grimbizis Grigoris
Published: January 2022

Background and Objectives: Ankylosing spondylitis (AS) is a condition that affects 0.1% to 0.5% of the adult population. The aim of this case report was to investigate the possible effects of the drugs taken for treatment of AS as well as mRNA vaccination for COVID-19 on semen quality by performing a highly detailed analysis. Materials and Methods: Sperm characteristics were examined by light microscopy, DNA fragmentation (DFI) was analysed by flow cytometry and morphology was evaluated by transmission electron microscopy (TEM). Results: Semen analysis under therapy with (1) celecoxib and sulphasalazine showed: concentration 47 million/mL, 53% progressive motility, 7% normal morphology and 9.6% DFI, (2) Golimumab and before mRNA Vaccination showed: concentration 108 million/mL, 82% progressive motility, 1% normal morphology and 7.6% DFI, and (3) Golimumab and after 3 doses of mRNA Vaccination showed: concentration 142 million/mL, 85% progressive motility, 1% normal morphology and 6.8% DFI. TEM revealed head, neck and tail abnormalities, as well as the presence of cells with incomplete spermiogenesis white cells and phagocytes in the sample under therapy with celecoxib and sulphasalazine. Golimumab treatment lead to an increased incidence of elongated heads but in general reduced inflammation as no white cells were evident in TEM. Conclusion: The anti-inflamatory drugs celecoxib and sulphasalazine had no adverse effect on sperm quality as all parameters were within normal limits and the patient achieved under that treatment 2 pregnancies following natural conception that lead to the birth of a healthy boy and girl respectively. Anti-TNFa treatment with Golimumab exerted a negative effect on morphology but not on concentration, motility and DFI. After 3 doses of mRNA Vaccination, sperm concentration increased while motility, morphology and DFI remained similar to the values before vaccination suggesting no negative effect of the mRNA vaccine for COVID-19 on sperm quality.
Combined Positive Score for Programmed Death Ligand-1 Expression and Inflammatory Microenvironment in Gastrointestinal Stromal Tumors
Authors: Herlea Vlad , Roșulescu Alexandra , Calotă Violeta Claudia , Croitoru Vlad , Stoica Mustafa Elena , Vasilescu Cătălin , Alexandrescu Sorin , Dumitrașcu Traian , Popescu Irinel , Dima Simona Olimpia , Sajin Maria
Published: January 2022

Background and Objectives: GISTs are the most frequent type of mesenchymal neoplasm of the digestive tract. The prognosis is mainly determined by tumor dimensions, mitotic rate and location, but other less well-documented factors can influence evolution and survival. The immune microenvironment and checkpoint molecule expression were proven to impact the prognosis in different types of cancer. The aim of this study was to determine PD-L1 expression in GISTs and to evaluate the level of intratumoral immune infiltration in relation to prognostic variables and survival. Materials and Methods: Sixty-five GISTs diagnosed in the same institution between 2015 and 2018 were immunohistochemically tested for PD-L1 and evaluated using CPS. Immune cells were emphasized, with CD3, CD4, CD8, CD20 and CD68 antibodies and quantified. All data were processed using statistical tools. Results: The median age was 61 years (range, 28–78) and 36 patients (55.4%) were males. The location of the tumors was predominantly gastric (46%), followed by the small bowel (17%) and colorectal (6%). In addition, 11% were EGISTs and 20% were secondary tumors (11% metastases and 9% local recurrences). PD-L1 had a variable expression in tumor and inflammatory cells, with a CPS ranging from 0 to 100. Moreover, 64.6% of cases were PD-L1 positive with no significant differences among categories of variables, such as the age and the sex of the patient, tumor location, the primary or secondary character of the tumor, dimensions, mitotic rate, the risk of disease progression and tumor cell type. Immune cells had a variable distribution throughout the tumors. CD3+ lymphocytes were the most frequent type. CD20+ cells were identified in a larger number in tumors ≤5 cm (p = 0.038). PD-L1-positive tumors had a higher number of immune cells, particularly CD3+, CD20+ and CD68+, in comparison to PD-L1-negative ones (p = 0.032, p = 0.051, p = 0.008). Epithelioid and mixed cell-type tumors had a higher number of CD68+ cells. Survival was not influenced by PD-L1 expression; instead, it was decreased in multifocal tumors (p = 0.0001) and in cases with Ki67 ≥ 50% (p = 0.008). Conclusions: PD-L1-positive expression and the presence of different immune cell types, in variable quantities, can contribute to a better understanding of the complex interactions between tumor cells and the microenvironment, with a possible therapeutic role in GISTs.
The Sex Difference in 6-h Ultra-Marathon Running—The Worldwide Trends from 1982 to 2020
Authors: Knechtle Beat , Weiss Katja , Villiger Elias , Scheer Volker , Gomes Thayse Natacha , Gajda Robert , Ouerghi Nejmeddine , Chtourou Hamdi , Nikolaidis Pantelis T. , Rosemann Thomas , Thuany Mabliny
Published: January 2022

Background and Objectives: The 6-h ultra-marathon is the shortest time-limited ultra-marathon race, but little has been investigated regarding this race format. Previously, only the age of peak performance in the context of longer time-limited ultra-marathons was determined. The purpose of this study was to investigate the trends in 6-h ultra-marathon races from 1982 to 2020 for female and male ultra-runners, the participation and performance by countries, the age of peak performance, and the differences in performance regarding countries. Materials and Methods: The sample included 23,203 female ultra-runners, aged 18–83 years, and 87,264 male ultra-runners, aged 18–85 years, who were finishers in a 6-h ultra-marathon held between 1982 and 2020. The age of peak performance was tested using the Kruskal–Wallis test, followed by the Bonferroni Correction. The difference in performance by countries was verified using a linear regression model with the fastest runners from Russia in women, and Tunisia in men, used as reference. Results: Over the years, the men-to-women ratio decreased. The mean age was 43.20 ± 9.30 years for female and 46.09 ± 10.17 years for male runners. Athletes in younger age groups were faster than athletes in older age groups. Most female and male participants originated from Germany. Women from Russia (10.01 ± 1.28 km/h) and men from Tunisia (12.16 ± 1.46 km/h) were the fastest. Conclusions: In summary, in 6-h ultra-marathons held between 1982 and 2020, the participation for both women and men increased, while the men-to-women ratio decreased. The mean age was higher in men compared to women. Most female and male runners originated from Germany, but the fastest women were from Russia, and the fastest men from Tunisia. Future studies need to investigate whether Russian women and Tunisian men are also the best in other distance-limited ultra-marathon races, such as 12-h and 24-h.
The Prevalence of Potential Drug-Drug Interactions in CKD-A Retrospective Observational Study of Cerrahpasa Nephrology Unit
Authors: Shahzadi Andleeb , Sonmez Ikbal , Kose Cagla , Oktan Burhaneddin , Alagoz Selma , Sonmez Haktan , Hussain Adil , Akkan Ahmet Gokhan
Published: January 2022

Background and Objectives: Chronic kidney disease (CKD) is usually linked with polypharmacy and patients are invariably at risk of complex medication regimens. The present study was designed to estimate the potential drug-drug interactions (pDDIs) through the prescription patterns provided to patients of the Nephrology Transplant Unit of Cerrahpasa Medical Faculty patients. Materials and Methods: 96 patients were included in the study. pDDIs among every combination of the prescribed drug were analyzed using the Thomson Reuters Micromedex. Results: We found 149 pDDIs making 2.16 interactions per prescription with incidence rates of 69.7%. Approximately 4.1% of interactions were of significant severity, 75.1% moderate severity, and 20.8% were classified as minor pDDIs. The most frequent interactions were found between iron and aluminum, calcium or magnesium-containing products (21.37%), calcium channel blockers and beta-blockers (8.96%); and aspirin and aluminum, calcium, or magnesium-containing products (7.58%). We identified 42 drug pairs with probability of clinical significance. The most commonly reported clinical outcomes of the pDDIs were hypo- or hypertension (39.24%), decreased drug efficacy (24.05%), and arrhythmia (9.49%). Aluminum, calcium, or magnesium-containing drug products (33.10%) constituted the primary class of drugs involved in interactions. Conclusions: This study showed pharmacodynamics (49%), pharmacokinetics (42.94%) interactions, polypharmacy and gender as determinant of pDDIs. A comprehensive multicenter research is required to decrease the morbidity and ease the state burden.
Splenectomy for Visceral Leishmaniasis Out of an Endemic Region: A Case Report and Literature Review
Authors: Lekic Nebojsa , Tadic Boris , Djordjevic Vladimir , Basaric Dragan , Micev Marjan , Vucelic Dragica , Mitrovic Milica , Grubor Nikola
Published: January 2022

Visceral leishmaniasis (also known as kala-azar) is characterized by fever, weight loss, swelling of the spleen and liver, and pancytopenia. If it is not treated, the fatality rate in developing countries can be as high as 100% within 2 years. In a high risk situation for perioperative bleeding due to severe thrombocytopenia/coagulopathy, we present a rare challenge for urgent splenectomy in a patient with previously undiagnosed visceral leishmaniasis. A histologic examination of the spleen revealed a visceral leishmaniasis, and the patient was successfully treated with amphotericin B.
Effective Balloon Pulmonary Angioplasty in a Patient with Chronic Thromboembolic Complications after Ventriculoatrial Shunt for Hydrocephalus in von Hippel–Lindau Disease
Authors: Gościniak Piotr , Larysz Michał , Sagan Leszek , Larysz Barbara , Syrenicz Anhelli , Kurzyna Marcin
Published: January 2022

Von Hippel–Lindau (VHL) disease along with chronic thromboembolic pulmonary hypertension (CTEPH) is a unique and unusual severe complication of ventriculoatrial (VA) shunt implantation in the treatment of hydrocephalus. To the best of our knowledge, this can be the first reported case of an effective treatment with balloon pulmonary angioplasty in a patient with VHL after VA shunt placement. The patient underwent six balloon pulmonary angioplasty procedures. All invasive procedures resulted in haemodynamic and functional improvement.
Temporary Trends Concerning the Extent and Efficacy of Atrial Fibrillation Ablation Using Radiofrequency Energy in a Polish Single-Center Experience
Authors: Myrda Krzysztof , Buchta Piotr , Błachut Aleksandra , Skrzypek Michał , Gąsior Mariusz
Published: January 2022

Background and Objectives: Atrial fibrillation (AF) is the most common supraventricular arrhythmia. Currently, catheter ablation is a preferred treatment strategy. The main objective of our study was a temporary trends analysis of patients’ data undergoing a single AF ablation procedure using radiofrequency energy (RF). The efficacy of the procedure underwent assessment during a 12-month follow-up. Materials and Methods: We analyzed 585 consecutive patients with symptomatic, recurrent, and drug-refractory AF hospitalized in our department between 2013 and 2018 who underwent RF ablation supported by a 3D electroanatomical system. The baseline characteristics, periprocedural parameters, and efficacy of the procedure at 6-, 9- and 12-month follow-ups were analyzed over the years. Results: The number of patients undergoing ablation increased. Patients with paroxysmal AF predominated (71.5%). However, the number of patients with the persistent type of arrhythmia increased over the years. The percentage of patients with chronic heart failure (CHF) increased to 27.5% in 2018, and patients presented with increasingly larger left atria (LA). In all patients, circumferential pulmonary vein isolation was performed. The percentage of patients who underwent arrhythmogenic substrate modification and cavotricuspid isthmus ablation increased. Over the years, the efficacy of a single procedure at the 12-month follow-up remained without significant differences between the years (72.0%, 69.6%, 75.5%, 74.8%, 71.7%, 71.7%). Conclusions: The rate of patients with CHF and advanced LA disease undergoing more extensive ablation increased over the years. The efficacy of a single procedure remained without significant differences between the years.
Evaluation of Dose-Dependent Obesity and Diabetes-Related Complications of Water Chestnut (Fruit of Trapa japonica) Extracts in Type II Obese Diabetic Mice Induced by 45% Kcal High-Fat Diet
Authors: Kang Hyun-Gu , Bashir Khawaja Muhammad Imran , Kim Ki-Young , Shin Su , Choi Min-Woo , Hong Eun-Jin , Choi Seong-Hun , Kim Joo-Wan , Choi Jae-Suk , Ku Sae-Kwang
Published: January 2022

Background and Objectives: The currently used pharmacological agents for metabolic disorders such as type II diabetes have several limitations and adverse effects; thus, there is a need for alternative therapeutic drugs and health functional foods. Materials and Methods: This study investigated the pharmacological effects of water chestnut (fruit of Trapa japonica) extracts (WC: 50–200 mg/kg) for type II diabetes using a 45% Kcal high-fat diet (HFD)-fed type II obese diabetic mice model for a period of 84 days, and the effects were compared to those of metformin (250 mg/kg). Results: Increases in body weight, serum biochemical indices such as triglycerides, low-density lipoprotein, and blood urea nitrogen, increases in antioxidant defense system enzymes such as catalase, superoxide dismutase, and glutathione, and mRNA expressions (such as AMPKα1 and AMPKα2) in the liver tissue and mRNA expressions (such as AMPKα2 mRNA, leptin, and C/EBPα) in the adipose tissue were observed in the HFD control group. The WC (50 mg/kg)-administered group showed no significant improvements in diabetic complications. However, HFD-induced obesity and diabetes-related complications such as hyperlipidemia, diabetic nephropathy, nonalcoholic fatty liver disease (NAFLD), oxidative stress, activity of antioxidant defense systems, and gene expressions were significantly and dose-dependently inhibited and/or normalized by oral administration of WC (100 mg/kg and 200 mg/kg), particularly at a dose of 100 mg/kg. Conclusions: The results of this study suggest that WC at an appropriate dose could be used to develop an effective therapeutic drug or functional food for type II diabetes and various associated complications, including NAFLD.
In Vitro and In Silico Antimalarial Evaluation of FM-AZ, a New Artemisinin Derivative
Authors: Tsamesidis Ioannis , Mousavizadeh Farnoush , Egwu Chinedu , Amanatidou Dionysia , Pantaleo Antonella , Benoit-Vical Françoise , Reybier Karine , Giannis Athanassios
Published: January 2022

Artemisinin-based Combination Therapies (ACTs) are currently the frontline treatment against Plasmodium falciparum malaria, but parasite resistance to artemisinin (ART) and its derivatives, core components of ACTs, is spreading in the Mekong countries. In this study, we report the synthesis of several novel artemisinin derivatives and evaluate their in vitro and in silico capacity to counteract Plasmodium falciparum artemisinin resistance. Furthermore, recognizing that the malaria parasite devotes considerable resources to minimizing the oxidative stress that it creates during its rapid consumption of hemoglobin and the release of heme, we sought to explore whether further augmentation of this oxidative toxicity might constitute an important addition to artemisinins. The present report demonstrates, in vitro, that FM-AZ, a newly synthesized artemisinin derivative, has a lower IC50 than artemisinin in P. falciparum and a rapid action in killing the parasites. The docking studies for important parasite protein targets, PfATP6 and PfHDP, complemented the in vitro results, explaining the superior IC50 values of FM-AZ in comparison with ART obtained for the ART-resistant strain. However, cross-resistance between FM-AZ and artemisinins was evidenced in vitro.
The Study of Ion Transport Parameters in MC-Based Electrolyte Membranes Using EIS and Their Applications for EDLC Devices
Authors: Aziz Shujahadeen , Dannoun Elham , Abdulwahid Rebar , Kadir Mohd , Nofal Muaffaq , Al-Saeedi Sameerah , Murad Ary
Published: January 2022

A solution cast technique was utilized to create a plasticized biopolymer-based electrolyte system. The system was prepared from methylcellulose (MC) polymer as the hosting material and potassium iodide (KI) salt as the ionic source. The electrolyte produced with sufficient conductivity was evaluated in an electrochemical double-layer capacitor (EDLC). Electrolyte systems’ electrical, structural, and electrochemical properties have been examined using various electrochemical and FTIR spectroscopic techniques. From the electrochemical impedance spectroscopy (EIS), a maximum ionic conductivity of 5.14 × 10−4 S cm−1 for the system with 50% plasticizer was recorded. From the EEC modeling, the ion transport parameters were evaluated. The extent of interaction between the components of the prepared electrolyte was investigated using Fourier transformed infrared spectroscopy (FTIR). For the electrolyte system (MC-KI-glycerol), the tion and electrochemical windows were 0.964 and 2.2 V, respectively. Another electrochemical property of electrolytes is transference number measurement (TNM), in which the ion predominantly responsibility was examined in an attempt to track the transport mechanism. The non-Faradaic nature of charge storing was proved from the absence of a redox peak in the cyclic voltammetry profile (CV). Several decisive parameters have been specified, such as specific capacitance (Cs), coulombic efficiency (η), energy density (Ed), and power density (Pd) at the first cycle, which were 68 F g−1, 67%, 7.88 Wh kg−1, and 1360 Wh kg−1, respectively. Ultimately, during the 400th cycle, the series resistance ESR varied from 70 to 310 ohms.
Effect of Vacancies on Dynamic Response and Spallation in Single-Crystal Magnesium by Molecular Dynamic Simulation
Authors: Jiang Chenying , Jian Zhiyong , Xiao Shifang , Li Xiaofan , Wang Kun , Deng Huiqiu , Hu Wangyu
Published: January 2022

The effect of vacancies on dynamic response and spallation in single-crystal magnesium (Mg) is investigated by nonequilibrium molecular dynamics simulations. The initial vacancy concentration (Cv) ranges from 0% to 2.0%, and the shock loading is applied along [0001] and [10–10] directions. The simulation results show that the effects of vacancy defects are strongly dependent on the shock directions. For shock along the [0001] direction, vacancy defects have a negligible effect on compression-induced plasticity, but play a role in increasing spall damage. In contrast, for shock along the [10–10] orientation, vacancy defects not only provide the nucleation sites for compression-induced plasticity, which mainly involves crystallographic reorientation, phase transition, and stacking faults, but also significantly reduce spall damage. The degree of spall damage is probably determined by a competitive mechanism between energy absorption and stress attenuation induced by plastic deformation. Void evolution during spallation is mainly based on the emission mechanism of dislocations. The {11–22} <11–23> pyramidal dislocation facilitates the nucleation of void in the [0001] shock, as well as the {1–100} <11–20> prismatic dislocation in the [10–10] shock. We also investigated the variation of spall strength between perfect and defective Mg at different shock velocities. The relevant results can provide a reference for future investigations on spall damage.
Effects of the Protein Concentration and Quality in a Canned Diet on the Fecal Microbiota of Healthy Adult Cats
Authors: Paßlack Nadine , Thies Louisa Verena , Vahjen Wilfried , Zentek Jürgen
Published: January 2022

Feline diets can markedly differ in their protein concentration and quality, which might also affect the intestinal microbiota of cats. In the present study, 6 canned diets, differing in their protein quality (high/low, achieved by varying amounts of meat and collagen-rich ingredients) and concentration (high quality/low quality: 36.2/36.7% in dry matter (DM), 43.3/45.0% in DM and 54.9/56.1% in DM), were fed to 10 healthy adult cats for 6 weeks each. At the end of the feeding periods, fecal samples were collected to analyze the microbiota (16S rDNA sequencing) and bacterial metabolites. Increasing dietary protein concentrations increased the relative abundance of Fusobacterium and Bacteroides as well as the concentrations of ammonium and n-valerate in the feces of the cats, independently of the dietary protein quality. A lower dietary protein quality was accompanied by a higher evenness index and a higher relative abundance of Fusobacteria and Bacteroidetes in the feces when compared to the feeding of the high protein quality diets. A promotion of bacterial proteolytic activity and, in particular, increased intestinal ammonium concentrations might be undesired effects of high protein intakes in cats. Whether the long-term feeding of those diets could be critical for feline health requires further investigation.
Bioinformatics Modelling and Metabolic Engineering of the Branched Chain Amino Acid Pathway for Specific Production of Mycosubtilin Isoforms in Bacillus subtilis
Authors: Guez Jean-Sébastien , Coucheney Françoise , Guy Joany , Béchet Max , Fontanille Pierre , Chihib Nour-Eddine , Niehren Joachim , Coutte François , Jacques Philippe
Published: January 2022

Mycosubtilin belongs to the family of lipopeptides. Different isoforms with various antifungal activities can be obtained according to the length and the isomery of the fatty acid. In this work, the activities of the mycosubtilin isoforms were first studied against the pathogen Aspergillus niger, revealing the high activity of the anteiso-C17 isoform. Modification of the mycosubtilin isoform patterns during cultures of the natural strain Bacillus subtilis ATCC 6633 was then investigated through amino acid feeding experiments. In parallel, single-gene knockouts and single-gene overexpression, leading to the overproduction of the anteiso-C15 fatty acid chains, were predicted using informatics tools which provide logical reasoning with formal models of reaction networks. In this way, it was in silico predicted that the single overexpression of the ilvA gene as well as the single knockout of the codY gene may lead to the overproduction of anteiso-C15 fatty acid chains. For the first time, it has been demonstrated that overexpression of ilvA helps to enhance the furniture of odd anteiso fatty acids leading to a favored mycosubtilin anteiso-C17 production pattern (+41%). Alternatively, a knock-out codY mutant led to a higher furniture of even iso fatty acids, leading to a favored mycosubtilin iso-C16 production pattern (+180%). These results showed that increased selective synthesis of particular isoforms of mycosubtilin through metabolic engineering is feasible, disclosing the interest of these approaches for future development of lipopeptide-producing strains.
A Metabolomics-Based Screening Proposal for Colorectal Cancer
Authors: Troisi Jacopo , Tafuro Maria , Lombardi Martina , Scala Giovanni , Richards Sean M. , Symes Steven J. K. , Ascierto Paolo Antonio , Delrio Paolo , Tatangelo Fabiana , Buonerba Carlo , Pierri Biancamaria , Cerino Pellegrino
Published: January 2022

Colorectal cancer (CRC) is a high incidence disease, characterized by high morbidity and mortality rates. Early diagnosis remains challenging because fecal occult blood screening tests have performed sub-optimally, especially due to hemorrhoidal, inflammatory, and vascular diseases, while colonoscopy is invasive and requires a medical setting to be performed. The objective of the present study was to determine if serum metabolomic profiles could be used to develop a novel screening approach for colorectal cancer. Furthermore, the study evaluated the metabolic alterations associated with the disease. Untargeted serum metabolomic profiles were collected from 100 CRC subjects, 50 healthy controls, and 50 individuals with benign colorectal disease. Different machine learning models, as well as an ensemble model based on a voting scheme, were built to discern CRC patients from CTRLs. The ensemble model correctly classified all CRC and CTRL subjects (accuracy = 100%) using a random subset of the cohort as a test set. Relevant metabolites were examined in a metabolite-set enrichment analysis, revealing differences in patients and controls primarily associated with cell glucose metabolism. These results support a potential use of the metabolomic signature as a non-invasive screening tool for CRC. Moreover, metabolic pathway analysis can provide valuable information to enhance understanding of the pathophysiological mechanisms underlying cancer. Further studies with larger cohorts, including blind trials, could potentially validate the reported results.
Low Cost, Easily-Assembled Centrifugal Buoyancy-Based Emulsification and Digital PCR
Authors: Zhou Wuping , Liu Cong , Zhang Tao , Jiang Keming , Li Haiwen , Zhang Zhiqiang , Tang Yuguo
Published: January 2022

Microfluidic-based droplet generation approaches require the design of microfluidic chips and a precise lithography process, which require skilled technicians and a long manufacturing time. Here we developed a centrifugal buoyancy-based emulsification (CBbE) method for producing droplets with high efficiency and minimal fabrication time. Our approach is to fabricate a droplet generation module that can be easily assembled using syringe needles and PCR tubes. With this module and a common centrifuge, high-throughput droplet generation with controllable droplet size could be realized in a few minutes. Experiments showed that the droplet diameter depended mainly on centrifugal speed, and droplets with controllable diameter from 206 to 158 μm could be generated under a centrifugal acceleration range from 14 to 171.9 g. Excellent droplet uniformity was achieved (CV < 3%) when centrifugal acceleration was greater than 108 g. We performed digital PCR tests through the CBbE approach and demonstrated that this cost-effective method not only eliminates the usage of complex microfluidic devices and control systems but also greatly suppresses the loss of materials and cross-contamination. CBbE-enabled droplet generation combines both easiness and robustness, and breaks the technical challenges by using conventional lab equipment and supplies.
A Novel Mini-Channel Heat Sink Design with Arc-Type Design Domain by Topology Optimization
Authors: Zhou Tao , Guo Chao , Shao Xiaodong
Published: January 2022

Based on the advantages of the topology optimization method, a new mini-channel heat sink with arc-type design domain topology design is proposed in this work. This arc-type design domain is used to realize the flow distribution uniformity. Two dual objective optimization functions were selected to complete the topology design, and two topology optimization mini-channel models M1 and M2 were obtained. The aim of M1 is to achieve minimum average temperature and fluid dissipation of the heat source area. The fluid dissipation was used to characterize the pressure drop characteristics. The aim of M2 is to achieve minimum temperature difference and fluid dissipation of the heat source area. Then, the fluid and heat transfer characteristics of M1, M2, and the traditional straight heat sink M3 were analyzed by numerical simulation. Compared with straight mini-channel heat sink M3, the temperature differences of the mini-channel heat sink designed M1 and the mini-channel heat sink designed M2 were decreased by 31.6% and 42.48%, respectively. Compared with M3, the pressure drops of M1 and M2 were decreased by 22.7% and 30.9%, respectively. Moreover, the Nusselt number of the mini-channel heat sink designed M1 was increased by 34.43%. In comparison, that of the mini-channel heat sink-designed M2 increased by 15.86%. The thermal performance evaluation criteria (PEC) showed that the PEC value of M1 was greater than 1.4, while the PEC value of M2 was less than 1.14. Finally, experiments were conducted for M1 to verify the accuracy of the numerical simulation. It was found that the simulation results agreed well with the experimental results.
Phylogenetic Revision and Patterns of Host Specificity in the Fungal Subphylum Entomophthoromycotina
Authors: Möckel Lars , Meusemann Karen , Misof Bernhard , Schwartze Volker U. , De Fine Licht Henrik H. , Voigt Kerstin , Stielow Benjamin , de Hoog Sybren , Beutel Rolf G. , Buellesbach Jan
Published: January 2022

The Entomophthoromycotina, a subphylum close to the root of terrestrial fungi with a bias toward insects as their primary hosts, has been notoriously difficult to categorize taxonomically for decades. Here, we reassess the phylogeny of this group based on conserved genes encoding ribosomal RNA and RNA polymerase II subunits, confirming their general monophyly, but challenging previously assumed taxonomic relationships within and between particular clades. Furthermore, for the prominent, partially human-pathogenic taxon Conidiobolus, a new type species C. coronatus is proposed in order to compensate for the unclear, presumably lost previous type species C. utriculosus Brefeld 1884. We also performed an exhaustive survey of the broad host spectrum of the Entomophthoromycotina, which is not restricted to insects alone, and investigated potential patterns of co-evolution across their megadiverse host range. Our results suggest multiple independent origins of parasitism within this subphylum and no apparent co-evolutionary events with any particular host lineage. However, Pterygota (i.e., winged insects) clearly constitute the most dominantly parasitized superordinate host group. This appears to be in accordance with an increased dispersal capacity mediated by the radiation of the Pterygota during insect evolution, which has likely greatly facilitated the spread, infection opportunities, and evolutionary divergence of the Entomophthoromycotina as well.
Host Cyanobacteria Killing by Novel Lytic Cyanophage YongM: A Protein Profiling Analysis
Authors: Zhang Shanshan , Zhao Baohua , Li Jing , Song Xiaofei , Tong Yigang , An Wenlin
Published: January 2022

Cyanobacteria are autotrophic prokaryotes that can proliferate robustly in eutrophic waters through photosynthesis. This can lead to outbreaks of lake “water blooms”, which result in water quality reduction and environmental pollution that seriously affect fisheries and aquaculture. The use of cyanophages to control the growth of cyanobacteria is an important strategy to tackle annual cyanobacterial blooms. YongM is a novel lytic cyanophage with a broad host spectrum and high efficiency in killing its host, cyanobacteria FACHB-596. However, changes in cyanophage protein profile during infestation and killing of the host remains unknown. To characterize the proteins and its regulation networks involved in the killing of host cyanobacteria by YongM and evaluate whether this strain YongM could be used as a chassis for further engineering to be a powerful tool in dealing with cyanobacterial blooms, we herein applied 4D label-free high-throughput quantitative proteomics to analyze differentially expressed proteins (DEPs) involved in cyanobacteria host response infected 1 and 8 h with YongM cyanophage. Metabolic pathways, such as photosynthesis, photosynthesis-antennal protein, oxidative phosphorylation, ribosome, carbon fixation, and glycolysis/glycol-isomerization were significantly altered in the infested host, whereas DEPs were associated with the metabolic processes of photosynthesis, precursor metabolites, energy production, and organic nitrogen compounds. Among these DEPs, key proteins involved in YongM-host interaction may be photosystem I P700 chlorophyll-a apolipoprotein, carbon dioxide concentration mechanism protein, cytochrome B, and some YongM infection lysis-related enzymes. Our results provide comprehensive information of protein profiles during the invasion and killing of host cyanobacteria by its cyanophage, which may shed light on future design and manipulation of artificial cyanophages against water blooms.
To Be or Not to Be an OXA-48 Carbapenemase
Authors: Dabos Laura , Oueslati Saoussen , Bernabeu Sandrine , Bonnin Rémy A. , Dortet Laurent , Naas Thierry
Published: January 2022

Since the first description of OXA-48, more than forty variants have been recovered from Enterobacterales isolates. Whereas some OXA-48-related enzymes have been reported as conferring similar resistance patterns, namely, the hydrolysis of carbapenems and penicillins with very weak or almost no activity against expanded-spectrum cephalosporins, some have reduced carbapenem and temocillin hydrolysis, and others hydrolyze expanded-spectrum cephalosporins and carbapenems only marginally. With such drastic differences in the hydrolytic profile, especially of carbapenems, it becomes urgent to establish hydrolytic cutoffs in order to determine when an OXA-48-like enzyme may be considered as a carbapenemase or not. With this aim, the coefficient of activity for imipenem (kcat/Km) was determined for a total of 30 enzymes, including OXA-48, OXA-48-like natural variants, and OXA-48 synthetic mutants. In addition, six different methods for the detection of carbapenemase-producers were performed. The coefficients of activity for imipenem for all the different enzymes went from 550 mM−1·s−1 to 0.02 mM−1·s−1. In order to match the coefficient of activity results with the biochemical confirmatory tests, we suggest the value of 0.27 mM−1·s−1 as the cutoff above which an OXA-48 variant may be considered a carbapenem-hydrolyzing enzyme.
Recent Advancements in Tracking Bacterial Effector Protein Translocation
Authors: Braet Julie , Catteeuw Dominiek , Van Damme Petra
Published: January 2022

Bacteria-host interactions are characterized by the delivery of bacterial virulence factors, i.e., effectors, into host cells where they counteract host immunity and exploit host responses allowing bacterial survival and spreading. These effectors are translocated into host cells by means of dedicated secretion systems such as the type 3 secretion system (T3SS). A comprehensive understanding of effector translocation in a spatio-temporal manner is of critical importance to gain insights into an effector’s mode of action. Various approaches have been developed to understand timing and order of effector translocation, quantities of translocated effectors and their subcellular localization upon translocation into host cells. Recently, the existing toolset has been expanded by newly developed state-of-the art methods to monitor bacterial effector translocation and dynamics. In this review, we elaborate on reported methods and discuss recent advances and shortcomings in this area of tracking bacterial effector translocation.
Universal Virucidal Activity of Calcium Bicarbonate Mesoscopic Crystals That Provides an Effective and Biosafe Disinfectant
Authors: Kirisawa Rikio , Kato Rika , Furusaki Koichi , Onodera Takashi
Published: January 2022

We investigated the virucidal effects in solution of a new type of disinfectant, calcium bicarbonate mesoscopic crystals, designated CAC-717, against various types of virus. CAC-717 in solution is alkaline (pH 12.4) and has a self-electromotive force that generates pulsed electrical fields. Upon application to human skin, the pH of the solution becomes 8.4. CAC-717 contains no harmful chemicals and is thus non-irritating and harmless to humans and animals. Its virucidal effects were tested against six types of animal virus: enveloped double-strand (ds)-DNA viruses, non-enveloped ds-DNA viruses, non-enveloped single strand (ss)-DNA viruses, enveloped ss-RNA viruses, non-enveloped ss-RNA viruses, and non-enveloped ds-RNA viruses. The treatment resulted in a reduction in viral titer of at least 3.00 log10 to 6.38 log10. Fetal bovine serum was added as a representative organic substance. When its concentration was ≥20%, the virucidal effect of CAC-717 was reduced. Real-time PCR revealed that CAC-717 did not reduce the quantity of genomic DNA of most of the DNA viruses, but it greatly reduced that of the genomic RNA of most of the RNA viruses. CAC-717 may therefore be a useful biosafe disinfectant for use against a broad range of viruses.
Viral Metagenomic Data Analyses of Five New World Bat Species from Argentina: Identification of 35 Novel DNA Viruses
Authors: Bolatti Elisa M. , Viarengo Gastón , Zorec Tomaz M. , Cerri Agustina , Montani María E. , Hosnjak Lea , Casal Pablo E. , Bortolotto Eugenia , Di Domenica Violeta , Chouhy Diego , Allasia María Belén , Barquez Rubén M. , Poljak Mario , Giri Adriana A.
Published: January 2022

Bats are natural reservoirs of a variety of zoonotic viruses, many of which cause severe human diseases. Characterizing viruses of bats inhabiting different geographical regions is important for understanding their viral diversity and for detecting viral spillovers between animal species. Herein, the diversity of DNA viruses of five arthropodophagous bat species from Argentina was investigated using metagenomics. Fecal samples of 29 individuals from five species (Tadarida brasiliensis, Molossus molossus, Eumops bonariensis, Eumops patagonicus, and Eptesicus diminutus) living at two different geographical locations, were investigated. Enriched viral DNA was sequenced using Illumina MiSeq, and the reads were trimmed and filtered using several bioinformatic approaches. The resulting nucleotide sequences were subjected to viral taxonomic classification. In total, 4,520,370 read pairs were sequestered by sequencing, and 21.1% of them mapped to viral taxa. Circoviridae and Genomoviridae were the most prevalent among vertebrate viral families in all bat species included in this study. Samples from the T. brasiliensis colony exhibited lower viral diversity than samples from other species of New World bats. We characterized 35 complete genome sequences of novel viruses. These findings provide new insights into the global diversity of bat viruses in poorly studied species, contributing to prevention of emerging zoonotic diseases and to conservation policies for endangered species.
The Effect of Recombinant Protein Production in Lactococcus lactis Transcriptome and Proteome
Authors: Monteiro Gabriel A. , Duarte Sofia O. D.
Published: January 2022

Lactococcus lactis is a food-grade, and generally recognized as safe, bacterium, which making it ideal for producing plasmid DNA (pDNA) or recombinant proteins for industrial or pharmaceutical applications. The present paper reviews the major findings from L. lactis transcriptome and proteome studies, with an overexpression of native or recombinant proteins. These studies should provide important insights on how to engineer the plasmid vectors and/or the strains in order to achieve high pDNA or recombinant proteins yields, with high quality standards. L. lactis harboring high copy numbers of plasmids for DNA vaccines production showed altered proteome profiles, when compared with a smaller copy number plasmid. For live mucosal vaccination applications, the cell-wall anchored antigens had shown more promising results, when compared with intracellular or secreted antigens. However, previous transcriptome and proteome studies demonstrated that engineering L. lactis to express membrane proteins, mainly with a eukaryotic background, increases the overall cellular burden. Genome engineering strategies could be used to knockout or overexpress the pinpointed genes, so as to increase the profitability of the process. Studies about the effect of protein overexpression on Escherichia coli and Bacillus subtillis transcriptome and proteome are also included.
A Polyphasic Approach Reveals Novel Genotypes and Updates the Genetic Structure of the Banana Fusarium Wilt Pathogen
Authors: Mostert Diane , Wicker Emmanuel , de Jager Mignon M. , Al Kaabi Saif M. , O’Neill Wayne T. , Perry Suzy , Li Chunyu , Ganyun Yi , Pegg Kenneth G. , Mostert Lizel , Viljoen Altus
Published: January 2022

Fusarium oxysporum f. sp. cubense (Foc) is a soil-borne fungus that causes Fusarium wilt, a destructive plant disease that has resulted in devastating economic losses to banana production worldwide. The fungus has a complex evolutionary history and taxonomic repute and consists of three pathogenic races and at least 24 vegetative compatibility groups (VCGs). Surveys conducted in Asia, Africa, the Sultanate of Oman and Mauritius encountered isolates of F. oxysporum pathogenic to banana that were not compatible to any of the known Foc VCGs. Genetic relatedness between the undescribed and known Foc VCGs were determined using a multi-gene phylogeny and diversity array technology (DArT) sequencing. The presence of putative effector genes, the secreted in xylem (SIX) genes, were also determined. Fourteen novel Foc VCGs and 17 single-member VCGs were identified. The multi-gene tree was congruent with the DArT-seq phylogeny and divided the novel VCGs into three clades. Clustering analysis of the DArT-seq data supported the separation of Foc isolates into eight distinct clusters, with the suite of SIX genes mostly conserved within these clusters. Results from this study indicates that Foc is more diverse than hitherto assumed.