bluuebunny/crossref_metadata_2025 huggingface dataset:
Metagenomics of Virus Diversities in Solid-State Brewing Process of Traditional Chinese Vinegar
Authors: Yu Zhen , Ma Yan , Guan Yingfen , Zhu Yuanyuan , Wang Ke , Wang Yuqin , Liu Peng , Chen Juan , Yu Yongjian
Published: October 2022

Traditional Chinese vinegar offers an exceptional flavor and rich nutrients due to its unique solid-state fermentation process, which is a multiple microbial fermentation system including various bacteria, fungi and viruses. However, few studies on the virus diversities in traditional Chinese vinegar have been reported. In this paper, using Zhenjiang aromatic vinegar as a model system, we systemically explored the viral communities in the solid-state brewing process of traditional Chinese vinegar using bacterial and viral metagenomes. Results showed that the viral diversity in vinegar Pei was extensive and the virus communities varied along with the fermentation process. In addition, there existed some interactions between viral and bacterial communities. Moreover, abundant antibiotic resistance genes were found in viromes, indicating that viruses might protect fermentation bacteria strains from the stress of antibiotics in the fermentation environment. Remarkably, we identified abundant auxiliary carbohydrate metabolic genes (including alcohol oxidases, the key enzymes for acetic acid synthesis) from viromes, implying that viruses might participate in the acetic acid synthesis progress of the host through auxiliary metabolic genes. Taken together, our results indicated the potential roles of viruses in the vinegar brewing process and provided a new perspective for studying the fermentation mechanisms of traditional Chinese vinegar.
Response Surface Modeling and Optimization of Enzymolysis Parameters for the In Vitro Antidiabetic Activities of Peanut Protein Hydrolysates Prepared Using Two Proteases
Authors: AL-Bukhaiti Wedad Q. , Al-Dalali Sam , Noman Anwar , Qiu Silin , Abed Sherif M. , Qiu Sheng-Xiang
Published: October 2022

Optimization of the enzymolysis process for preparing peanut protein hydrolysates using alcalase and trypsin was performed by employing the central composite design (CCD) of response surface methodology (RSM). The independent variables were solid-to-liquid ratio (S/L), enzyme-to-substrate ratio (E/S), pH, and reaction temperature, while the response variables were degree of hydrolysate (DH), α-amylase, and α-glucosidase inhibitory activity. The highest DH (22.84% and 14.63%), α-amylase inhibition (56.78% and 40.80%), and α-glucosidase inhibition (86.37% and 86.51%) were obtained under optimal conditions, which were S/L of 1:26.22 and 1:30 w/v, E/S of 6% and 5.67%, pH of 8.41 and 8.56, and temperature of 56.18 °C and 58.75 °C at 3 h using alcalase (AH) and trypsin (TH), respectively. Molecular weight distributions of peanut protein hydrolysates were characterized by SDS-PAGE, which were mostly ˂10 kDa for both hydrolysates. Lyophilized AH and TH had IC50 values of 6.77 and 5.86 mg/mL for α-amylase inhibitory activity, and 6.28 and 5.64 mg/mL for α-glucosidase inhibitory activity. The IC50 of AH and TH against DPPH radical was achieved at 4.10 and 3.20 mg/mL and against ABTS radical at 2.71 and 2.32 mg/mL, respectively. The obtained hydrolysates with antidiabetic activity could be utilized as natural alternatives to synthetic antidiabetics, particularly in food and pharmaceutical products.
Single, Subsequent, or Simultaneous Treatments to Mitigate Mycotoxins in Solid Foods and Feeds: A Critical Review
Authors: Abou Dib Alaa , Assaf Jean Claude , El Khoury André , El Khatib Sami , Koubaa Mohamed , Louka Nicolas
Published: October 2022

Mycotoxins in solid foods and feeds jeopardize the public health of humans and animals and cause food security issues. The inefficacy of most preventive measures to control the production of fungi in foods and feeds during the pre-harvest and post-harvest stages incited interest in the mitigation of these mycotoxins that can be conducted by the application of various chemical, physical, and/or biological treatments. These treatments are implemented separately or through a combination of two or more treatments simultaneously or subsequently. The reduction rates of the methods differ greatly, as do their effect on the organoleptic attributes, nutritional quality, and the environment. This critical review aims at summarizing the latest studies related to the mitigation of mycotoxins in solid foods and feeds. It discusses and evaluates the single and combined mycotoxin reduction treatments, compares their efficiency, elaborates on their advantages and disadvantages, and sheds light on the treated foods or feeds, as well as on their environmental impact.
Flaxseed Bioactive Compounds: Chemical Composition, Functional Properties, Food Applications and Health Benefits-Related Gut Microbes
Authors: Mueed Abdul , Shibli Sahar , Korma Sameh A. , Madjirebaye Philippe , Esatbeyoglu Tuba , Deng Zeyuan
Published: October 2022

Flaxseed (Linum usitatissimum L.) has gained worldwide recognition as a health food because of its abundance in diverse nutrients and bioactive compounds such as oil, fatty acids, proteins, peptides, fiber, lignans, carbohydrates, mucilage, and micronutrients. These constituents attribute a multitude of beneficial properties to flaxseed that makes its use possible in various applications, such as nutraceuticals, food products, cosmetics, and biomaterials. The importance of these flaxseed components has also increased in modern times because of the newer trend among consumers of greater reliance on a plant-based diet for fulfilling their nutritional requirements, which is perceived to be hypoallergenic, more environmentally friendly, sustainable, and humane. The role of flaxseed substances in the maintenance of a healthy composition of the gut microbiome, prevention, and management of multiple diseases has recently been elucidated in various studies, which have highlighted its importance further as a powerful nutritional remedy. Many articles previously reported the nutritive and health benefits of flaxseed, but no review paper has been published reporting the use of individual flaxseed components in a manner to improve the techno-functional properties of foods. This review summarizes almost all possible applications of flaxseed ingredients in food products from an extensive online literature survey; moreover, it also outlines the way forward to make this utilization even better.
Effect of Autochthonous Lactic Acid Bacteria-Enhanced Fermentation on the Quality of Suancai
Authors: Cao Xinying , Zhao Mingwei , Zou Sibo , Li Zhigao , Wu Yuzheng , Ji Chaofan , Chen Yingxi , Dong Liang , Zhang Sufang , Liang Huipeng
Published: October 2022

The lactic acid bacteria (LABs) used for fermentation have an extremely vital impact on the quality of Suancai, a fermented vegetable. The bacterial diversity and metabolites of inoculated Suancai with LABs, including Lactiplantibacillus plantarum (Lb. plantarum), Levilactabacillus brevis (Lb. brevis), and Leuconostoc mesenteroides (Leu. mesenteroides), were investigated. The inoculation of LABs significantly decreased the pH and the content of nitrite. The Suancai inoculated with LABs had a higher content of the total titratable acidity (TTA) and organic acids than spontaneous fermentation. The LABs inoculation significantly influenced the bacterial community structures, which directly or indirectly caused changes of metabolites. The bacterial community profiles of Suancai inoculated with Lb. plantarum were more similar to spontaneous fermentation. The inoculation of Lb. plantarum, Lb. brevis, and Leu. mesenteroides could increase its abundance in Suancai. Whatever the species inoculated, Lb. plantarum was always the predominant bacterium in Suancai after fermentation. The inoculated LABs were positively correlated with most volatile compounds and amino acids. The inoculated LABs significantly improved the volatile compounds and amino acid content of Suancai. This study could contribute to understanding the function of starters in Suancai fermentation and promote the selection of applicable starters for high-quality Suancai production.
Ready-to-Eat Fish Cake Processing Methods and the Impacts on Quality and Flavor
Authors: Jiang Caiyan , Chen Yao , Li Shuang , Shang Shan , Fu Baoshang , Wang Lina , Dong Xiuping , Jiang Pengfei
Published: October 2022

This study aimed to compare tilapia fish cake drying and sterilization conditions (105, 115, and 121 °C) on the quality of the cakes. The impacts of volatile flavor substances, the chroma value, quality and structure characteristics, microscopic structure, and the types and content of volatile flavor substances were also analyzed. The results showed that after drying and sterilization, the L* value, W value and delta-E value of fish cakes decreased significantly from 77.12 to 64.77, 66.21 to 52.57, 10.46 to 24.50, respectively. However, a* value and b* value increased significantly from 0.30 to 6.97 and 24.85 to 30.89, respectively. The elasticity, hardness, and chewiness increased significantly with the drying process but decreased significantly with the increased sterilization temperature. Scanning electron microscopy results showed that the internal pores of the fish cakes became smaller, and the tissue structure was closer after drying. Gas chromatography—ion mobile spectrometry analysis identified a total of 36 volatile flavor compounds. Among these, ketones comprised the largest content, aldehydes represented the largest variety, and all volatile compounds contributed significantly to the flavor of fish cake. PCA results and nearest-neighbor fingerprint analysis showed that there were obvious differences in volatile flavor compounds between different treatments. In summary, this study conducted a detailed comparative analysis of the quality and flavor of fish cakes subjected to different processing methods. These findings contribute suggestions for sterilization temperatures in industrial production processes.
Dietary Phenolic Compounds as Anticancer Natural Drugs: Recent Update on Molecular Mechanisms and Clinical Trials
Authors: Bakrim Saad , El Omari Nasreddine , El Hachlafi Naoufal , Bakri Youssef , Lee Learn-Han , Bouyahya Abdelhakim
Published: October 2022

Given the stochastic complexity of cancer diseases, the development of chemotherapeutic drugs is almost limited by problems of selectivity and side effects. Furthermore, an increasing number of protective approaches have been recently considered as the main way to limit these pathologies. Natural bioactive compounds, and particularly dietary phenolic compounds, showed major protective and therapeutic effects against different types of human cancers. Indeed, phenolic substances have functional groups that allow them to exert several anti-cancer mechanisms, such as the induction of apoptosis, autophagy, cell cycle arrest at different stages, and the inhibition of telomerase. In addition, in vivo studies show that these phenolic compounds also have anti-angiogenic effects via the inhibition of invasion and angiogenesis. Moreover, clinical studies have already highlighted certain phenolic compounds producing clinical effects alone, or in combination with drugs used in chemotherapy. In the present work, we present a major advance in research concerning the mechanisms of action of the different phenolic compounds that are contained in food medicinal plants, as well as evidence from the clinical trials that focus on them.
The Effect of Stabilized Rice Bran Addition on Physicochemical, Sensory, and Techno-Functional Properties of Bread
Authors: Espinales Cindy , Cuesta Adriana , Tapia Javier , Palacios-Ponce Sócrates , Peñas Elena , Martínez-Villaluenga Cristina , Espinoza Alexander , Cáceres Patricio
Published: October 2022

Rice bran (RB) is a valuable byproduct derived from rice milling that represents an excellent opportunity for dietary inclusion. Bioactive components with antioxidant potential have been reported in RB, gaining the considerable attention of researchers. However, RB requires a stabilization process after milling to prevent it from becoming rancid and promote its commercial consumption. The aim of this study was to evaluate the effects of substituting stabilized rice bran (SRB) for wheat flour at levels of 10, 15, 20 and 25% on the proximate composition, dietary fiber, dough rheology, antioxidant properties, content of bioactive compounds, and sensory attributes of white wheat-based bread. Results indicated that the incorporation of SRB increased the bread’s insoluble dietary fiber, phytic acid, total polyphenol content, γ-oryzanol, γ-aminobutyric acid, and antioxidant properties, while decreased its water absorption capacity, elasticity, volume, β-glucans, and soluble dietary fiber content. Moreover, substituting wheat flour for SRB at levels higher than 15% affected sensory attributes, such as color, odor, flavor, and softness. This study highlights the potential application of SRB flour in bread-making to increase nutritional, and functional properties of white wheat bread.
Characterization and Immunological Activity of Exopolysaccharide from Lacticaseibacillus paracasei GL1 Isolated from Tibetan Kefir Grains
Authors: Wang Xiaomeng , Tian Juanjuan , Zhang Xueliang , Tang Nanyu , Rui Xin , Zhang Qiuqin , Dong Mingsheng , Li Wei
Published: October 2022

Two exopolysaccharide fractions (GL1-E1 and GL1-E2) of Lacticaseibacillus paracasei GL1 were isolated with the molecular weights of 3.9 × 105 Da and 8.2 × 105 Da, respectively. Both fractions possessed mannose, glucose, and galactose in molar ratios of 1.16:1.00:0.1, and 3.81:1.00:0.12, respectively. A structural arrangement of two fractions was proposed by methylation, one-dimensional and two-dimensional nuclear magnetic resonance experiments. The backbone of GL1-E1 consisted of →4)-α-D-Glcp(1→, →3,4)-α-D-Manp(1→, →3,6)-α-D-Manp(1→, →6)-α-D-Manp(1→, and →6)-α-D-Galp(1→ with α-D-Glcp at branching point. The backbone of GL1-E2 consisted of →4)-α-D-Glcp(1→, →3,4)-α-D-Manp(1→, →3,6)-α-D-Manp(1→, →6)-α-D-Manp(1→, →6)-α-D-Galp(1→, and →4)-β-D-Manp(1→, and the side chain also consisted of α-D-Manp residue. In addition, the differential scanning calorimetry (DSC) analysis indicated that both GL1-E1 and GL1-E2 had good thermal stability. Furthermore, the two fractions could promote the viability of RAW264.7 cells and exert an immunomodulatory role by enhancing phagocytosis, increasing nitric oxide (NO) release and promoting the expression of cytokines.
Effects of Dietary L-malic Acid Supplementation on Meat Quality, Antioxidant Capacity and Muscle Fiber Characteristics of Finishing Pigs
Authors: Yan Enfa , Wang Yubo , He Linjuan , Guo Jianxin , Zhang Xin , Yin Jingdong
Published: October 2022

L-malic acid is a vital intermediate in the citric acid cycle and has been reported to improve the antioxidant capacity and aerobic oxidation of weaned piglets; however, its application in finishing pigs is limited at present. This study explored the effects of dietary L-malic acid supplementation on the carcass traits and meat quality of finishing pigs. In a 45-day experiment, 192 Duroc × Landrace × Yorkshire pigs (75.01 ± 0.51 kg) were divided into four treatments, i.e., a basal diet supplemented with 0, 0.5%, 1%, and 2% L-malic acid complex. The results showed that L-malic acid supplementation had no effects on the growth performance of finishing pigs. Importantly, L-malic acid significantly increased redness (a*) value at 24-h postmortem (quadratic, p < 0.05) and tended to increase the proportion of oxymyoglobin (OMb) (quadratic, p = 0.10), as well as the total antioxidant capacity (T-AOC) activity (quadratic, p = 0.08) in the longissimus dorsi (LD) muscle. Further, dietary supplementation of 1% L-malic acid markedly increased the protein expression level of slow skeletal myosin heavy chain (MyHC) in the LD muscle (p < 0.05). Moreover, 0.5% and 2% L-malic acid supplementation significantly increased carcass length and loin eye area (p < 0.05). In conclusion, dietary L-malic acid could effectively improve the meat color and carcass traits of finishing pigs.
Soliton Solution of the Peyrard–Bishop–Dauxois Model of DNA Dynamics with M-Truncated and β-Fractional Derivatives Using Kudryashov’s R Function Method
Authors: Wang Xiaoming , Akram Ghazala , Sadaf Maasoomah , Mariyam Hajra , Abbas Muhammad
Published: October 2022

In this paper, the Peyrard–Bishop–Dauxois model of DNA dynamics is discussed along with the fractional effects of the M-truncated derivative and β-derivative. The Kudryashov’s R method was applied to the model in order to obtain a solitary wave solution. The obtained solution is explained graphically and the fractional effects of the β and M-truncated derivatives are also shown for a better understanding of the model.
Modal Shifted Fifth-Kind Chebyshev Tau Integral Approach for Solving Heat Conduction Equation
Authors: Atta Ahmed Gamal , Abd-Elhameed Waleed Mohamed , Moatimid Galal Mahrous , Youssri Youssri Hassan
Published: October 2022

In this study, a spectral tau solution to the heat conduction equation is introduced. As basis functions, the orthogonal polynomials, namely, the shifted fifth-kind Chebyshev polynomials (5CPs), are used. The proposed method’s derivation is based on solving the integral equation that corresponds to the original problem. The tau approach and some theoretical findings serve to transform the problem with its underlying conditions into a suitable system of equations that can be successfully solved by the Gaussian elimination method. For the applicability and precision of our suggested algorithm, some numerical examples are given.
Diverse Polarimetric Features of AGN Jets from Various Viewing Angles: Towards a Unified View
Authors: Tsunetoe Yuh , Mineshige Shin , Kawashima Tomohisa , Ohsuga Ken , Akiyama Kazunori , Takahashi Hiroyuki R.
Published: October 2022

Here, we demonstrate that polarization properties show a wide diversity depending on viewing angles. To simulate images of a supermassive black hole and surrounding plasma, we performed a full-polarimetric general relativistic radiative transfer based on three-dimensional general relativistic magnetohydrodynamics models with moderate magnetic strengths. Under an assumption of a hot-jet and cold-disk in the electron temperature prescription, we confirmed a typical scenario where polarized synchrotron emissions from the funnel jet experience Faraday rotation and conversion in the equatorial disk. Further, we found that linear polarization vectors are inevitably depolarized for edge-on-like observers, whereas a portion of vectors survive and reach the observers in face-on-like cases. We also found that circular polarization components have persistent signs in the face-on cases, and changing signs in the edge-on cases. It is confirmed that these features are smoothly connected via intermediate viewing-angle cases. These results are due to Faraday rotation/conversion for different viewing angles, and suggest that a combination of linear and circular polarimetry can give a constraint on the inclination between the observer and black hole’s (and/or disk’s) rotating-axis and plasma properties in the jet–disk structure. These can also lead to a more statistical and unified interpretation for a diversity of emissions from active galactic nuclei.
Printing Structurally Anisotropic Biocompatible Fibrillar Hydrogel for Guided Cell Alignment
Authors: Chen Zhengkun , Khuu Nancy , Xu Fei , Kheiri Sina , Yakavets Ilya , Rakhshani Faeze , Morozova Sofia , Kumacheva Eugenia
Published: October 2022

Many fibrous biological tissues exhibit structural anisotropy due to the alignment of fibers in the extracellular matrix. To study the impact of such anisotropy on cell proliferation, orientation, and mobility, it is important to recapitulate and achieve control over the structure of man-made hydrogel scaffolds for cell culture. Here, we report a chemically crosslinked fibrous hydrogel due to the reaction between aldehyde-modified cellulose nanofibers and gelatin. We explored two ways to induce structural anisotropy in this gel by extruding the hydrogel precursor through two different printheads. The cellulose nanofibers in the hydrogel ink underwent shear-induced alignment during extrusion and retained it in the chemically crosslinked hydrogel. The degree of anisotropy was controlled by the ink composition and extrusion flow rate. The structural anisotropy of the hydrogel extruded through a nozzle affected the orientation of human dermal fibroblasts that were either seeded on the hydrogel surface or encapsulated in the extruded hydrogel. The reported straightforward approach to constructing fibrillar hydrogel scaffolds with structural anisotropy can be used in studies of the biological impact of tissue anisotropy.
Imaging Arctic Permafrost: Modeling for Choice of Geophysical Methods
Authors: Buddo Igor , Misyurkeeva Natalya , Shelokhov Ivan , Chuvilin Evgeny , Chernikh Alexey , Smirnov Alexander
Published: October 2022

Knowledge of permafrost structure, with accumulations of free natural gas and gas hydrates, is indispensable for coping with spontaneous gas emission and other problems related to exploration and production drilling in Arctic petroleum provinces. The existing geophysical methods have different potentialities for imaging the permafrost base and geometry, vertical fluid conduits (permeable zones), taliks, gas pockets, and gas hydrate accumulations in the continental Arctic areas. The synthesis of data on cryological and geological conditions was the basis for a geophysical–geological model of northern West Siberia to a depth of 400 m, which includes modern permafrost, lenses of relict permafrost with hypothetical gas hydrates, and a permeable zone that may be a path for the migration of gas–water fluids. The model was used to model synthetic seismic, electrical resistivity tomography (ERT), and transient electromagnetic (TEM) data, thus testing the advantages and drawbacks of the three methods. Electrical resistivity tomography has insufficient penetration to resolve all features and can run only in the summer season. Seismic surveys have limitations in mapping fluid conduits, though they can image a horizontally layered structure in any season. Shallow transient electromagnetic (sTEM) soundings can image any type of features included into the geological model and work all year round. Thus, the best strategy is to use TEM surveys as the main method, combined with seismic and ERT data. Each specific method is chosen proceeding from economic viability and feasibility in the specific physiographic conditions of mountain and river systems.
Hospital Staffing during the COVID-19 Pandemic in Sweden
Authors: Rosenbäck Ritva , Lantz Björn , Rosén Peter
Published: October 2022

Staff management challenges in the healthcare system are inherently different during pandemic conditions than under normal circumstances. Surge capacity must be rapidly increased, particularly in the intensive care units (ICU), to handle the increased pressure, without depleting the rest of the system. In addition, sickness or fatigue among the staff can become a critical issue. This study explores the lessons learned by first- and second-line managers in Sweden with regard to staff management during the COVID-19 pandemic. A mixed-methods approach was used, with preliminary qualitative interview (n = 38) and principal quantitative questionnaire (n = 272) studies, based on principal component and multiple regression analyses. The results revealed that the pandemic created four types of challenges relating to staff management: staff movement within hospitals; addition of external staff; addition of hours for existing staff through overtime and new shift schedules; and avoidance of lost hours due to sickness or fatigue. Furthermore, the effects of these managerial challenges were different in the first wave than in later waves, and they significantly differed between the ICU and other units. Therefore, a greater proactive focus on staff management would be beneficial in future pandemic situations.
Small Extracellular Vesicles from Hypoxic Triple-Negative Breast Cancer Cells Induce Oxygen-Dependent Cell Invasion
Authors: Pachane Bianca Cruz , Nunes Ana Carolina Caetano , Cataldi Thais Regiani , Micocci Kelli Cristina , Moreira Bianca Caruso , Labate Carlos Alberto , Selistre-de-Araujo Heloisa Sobreiro , Altei Wanessa Fernanda
Published: October 2022

Hypoxia, a condition of low oxygenation frequently found in triple-negative breast tumors (TNBC), promotes extracellular vesicle (EV) secretion and favors cell invasion, a complex process in which cell morphology is altered, dynamic focal adhesion spots are created, and ECM is remodeled. Here, we investigated the invasive properties triggered by TNBC-derived hypoxic small EV (SEVh) in vitro in cells cultured under hypoxic (1% O2) and normoxic (20% O2) conditions, using phenotypical and proteomic approaches. SEVh characterization demonstrated increased protein abundance and diversity over normoxic SEV (SEVn), with enrichment in pro-invasive pathways. In normoxic cells, SEVh promotes invasive behavior through pro-migratory morphology, invadopodia development, ECM degradation, and matrix metalloprotease (MMP) secretion. The proteome profiling of 20% O2-cultured cells exposed to SEVh determined enrichment in metabolic processes and cell cycles, modulating cell health to escape apoptotic pathways. In hypoxia, SEVh was responsible for proteolytic and catabolic pathway inducement, interfering with integrin availability and gelatinase expression. Overall, our results demonstrate the importance of hypoxic signaling via SEV in tumors for the early establishment of metastasis.
Using Computational Drug-Gene Analysis to Identify Novel Therapeutic Candidates for Retinal Neuroprotection
Authors: Xie Edward , Nadeem Urooba , Xie Bingqing , D’Souza Mark , Sulakhe Dinanath , Skondra Dimitra
Published: October 2022

Retinal cell death is responsible for irreversible vision loss in many retinal disorders. No commercially approved treatments are currently available to attenuate retinal cell loss and preserve vision. We seek to identify chemicals/drugs with thoroughly-studied biological functions that possess neuroprotective effects in the retina using a computational bioinformatics approach. We queried the National Center for Biotechnology Information (NCBI) to identify genes associated with retinal neuroprotection. Enrichment analysis was performed using ToppGene to identify compounds related to the identified genes. This analysis constructs a Pharmacome from multiple drug-gene interaction databases to predict compounds with statistically significant associations to genes involved in retinal neuroprotection. Compounds with known deleterious effects (e.g., asbestos, ethanol) or with no clinical indications (e.g., paraquat, ozone) were manually filtered. We identified numerous drug/chemical classes associated to multiple genes implicated in retinal neuroprotection using a systematic computational approach. Anti-diabetics, lipid-lowering medicines, and antioxidants are among the treatments anticipated by this analysis, and many of these drugs could be readily repurposed for retinal neuroprotection. Our technique serves as an unbiased tool that can be utilized in the future to lead focused preclinical and clinical investigations for complex processes such as neuroprotection, as well as a wide range of other ocular pathologies.
Upgrading Monocytes Therapy for Critical Limb Ischemia Patient Treatment: Pre-Clinical and GMP-Validation Aspects
Authors: Rusconi Giulio , Cusumano Giuseppe , Mariotta Luca , Canevascini Reto , Gola Mauro , Gornati Rosalba , Soldati Gianni
Published: October 2022

Advanced cell therapy medicinal products (ATMP) are at the forefront of a new range of biopharmaceuticals. The use of ATMP has evolved and increased in the last decades, representing a new approach to treating diseases that are not effectively managed with conventional treatments. The standard worldwide recognized for drug production is the Good Manufacturing Practices (GMP), widely used in the pharma production of synthesized drugs but applying also to ATMP. GMP guidelines are worldwide recognized standards to manufacture medicinal products to guarantee high quality, safety, and efficacy. In this report, we describe the pre-clinical and the GMP upgrade of peripheral blood mononuclear cell (PBMC) preparation, starting from peripheral blood and ending up with a GMP-grade clinical product ready to be used in patients with critical limb ischemia (CLI). We also evaluated production in hypoxic conditions to increase PBMC functional activity and angiogenic potential. Furthermore, we extensively analyzed the storage and transport conditions of the final product as required by the regulatory body for ATMPs. Altogether, results suggest that the whole manufacturing process can be performed for clinical application. Peripheral blood collected by a physician should be transported at room temperature, and PBMCs should be isolated in a clean room within 8 h of venipuncture. Frozen cells can be stored in nitrogen vapors and thawed for up to 12 months. PBMCs resuspended in 5% human albumin solution should be stored and transported at 4 °C before injection in patients within 24 h to thawing. Hypoxic conditioning of PBMCs should be implemented for clinical application, as it showed a significant enhancement of PBMC functional activity, in particular with increased adhesion, migration, and oxidative stress resistance. We demonstrated the feasibility and the quality of a GMP-enriched suspension of monocytes as an ATMP, tested in a clean room facility for all aspects related to production in respect of all the GMP criteria that allow its use as an ATMP. We think that these results could ease the way to the clinical application of ATMPs.
The Effect of Angiogenesis-Based Scaffold of MesoporousBioactive Glass Nanofiber on Osteogenesis
Authors: Zheng Weijia , Bai Zhenzu , Huang Shan , Jiang Kai , Liu Long , Wang Xiaoyan
Published: October 2022

There is still an urgent need for more efficient biological scaffolds to promote the healing of bone defects. Vessels can accelerate bone growth and regeneration by transporting nutrients, which is an excellent method to jointly increase osteogenesis and angiogenesis in bone regeneration. Therefore, we aimed to prepare a composite scaffold that could promote osteogenesis with angiogenesis to enhance bone defect repair. Here, we report that scaffolds were prepared by coaxial electrospinning with mesoporous bioactive glass modified with amino (MBG-NH2) adsorbing insulin-like growth factor-1 (IGF-1) as the core and silk fibroin (SF) adsorbing vascular endothelial growth factor (VEGF) as the shell. These scaffolds were named MBG-NH2/IGF@SF/VEGF and might be used as repair materials to promote bone defect repair. Interestingly, we found that the MBG-NH2/IGF@SF/VEGF scaffolds had nano-scale morphology and high porosity, as well as enough mechanical strength to support the tissue. Moreover, MBG-NH2 could sustain the release of IGF-1 to achieve long-term repair. Additionally, the MBG-NH2/IGF@SF/VEGF scaffolds could significantly promote the mRNA expression levels of osteogenic marker genes and the protein expression levels of Bmp2 and Runx2 in bone marrow mesenchymal stem cells (BMSCs). Meanwhile, the MBG-NH2/IGF@SF/VEGF scaffolds promoted osteogenesis by simulating Runx2 transcription activity through the phosphorylated Erk1/2-activated pathway. Intriguingly, the MBG-NH2/IGF@SF/VEGF scaffolds could also significantly promote the mRNA expression level of angiogenesis marker genes and the protein expression level of CD31. Furthermore, RNA sequencing verified that the MBG-NH2/IGF@SF/VEGF scaffolds had excellent performance in promoting bone defect repair and angiogenesis. Consistent with these observations, we found that the MBG-NH2/IGF@SF/VEGF scaffolds demonstrated a good repair effect on a critical skull defect in mice in vivo, which not only promoted the formation of blood vessels in the haversian canal but also accelerated the bone repair process. We concluded that these MBG-NH2/IGF@SF/VEGF scaffolds could promote bone defect repair under accelerating angiogenesis. Our finding provides a new potential biomaterial for bone tissue engineering.
Lipopolysaccharides (LPSs) as Potent Neurotoxic Glycolipids in Alzheimer’s Disease (AD)
Authors: Zhao Yuhai , Jaber Vivian R. , Pogue Aileen I. , Sharfman Nathan M. , Taylor Christopher , Lukiw Walter J.
Published: October 2022

Lipopolysaccharides (LPSs) are microbiome-derived glycolipids that are among the most potent pro-inflammatory neurotoxins known. In Homo sapiens, the major sources of LPSs are gastrointestinal (GI)-tract-resident facultative anaerobic Gram-negative bacilli, including Bacteroides fragilis and Escherichia coli. LPSs have been abundantly detected in aged human brain by multiple independent research investigators, and an increased abundance of LPSs around and within Alzheimer’s disease (AD)-affected neurons has been found. Microbiome-generated LPSs and other endotoxins cross GI-tract biophysiological barriers into the systemic circulation and across the blood–brain barrier into the brain, a pathological process that increases during aging and in vascular disorders, including ‘leaky gut syndrome’. Further evidence indicates that LPSs up-regulate pro-inflammatory transcription factor complex NF-kB (p50/p65) and subsequently a set of NF-kB-sensitive microRNAs, including miRNA-30b, miRNA-34a, miRNA-146a and miRNA-155. These up-regulated miRNAs in turn down-regulate a family of neurodegeneration-associated messenger RNA (mRNA) targets, including the mRNA encoding the neuron-specific neurofilament light (NF-L) chain protein. While NF-L has been reported to be up-regulated in peripheral biofluids in AD and other progressive and lethal pro-inflammatory neurodegenerative disorders, NF-L is significantly down-regulated within neocortical neurons, and this may account for neuronal atrophy, loss of axonal caliber and alterations in neuronal cell shape, modified synaptic architecture and network deficits in neuronal signaling capacity. This paper reviews and reveals the most current findings on the neurotoxic aspects of LPSs and how these pro-inflammatory glycolipids contribute to the biological mechanism of progressive, age-related and ultimately lethal neurodegenerative disorders. This recently discovered gut-microbiota-derived LPS–NF-kB–miRNA-30b–NF-L pathological signaling network: (i) underscores a direct positive pathological link between the LPSs of GI-tract microbes and the inflammatory neuropathology, disordered cytoskeleton, and disrupted synaptic-signaling of the AD brain and stressed human brain cells in primary culture; and (ii) is the first example of a microbiome-derived neurotoxic glycolipid having significant detrimental miRNA-mediated actions on the expression of NF-L, an abundant filamentous protein known to be important in the maintenance of neuronal and synaptic homeostasis.
Isomelezitose Overproduction by Alginate-Entrapped Recombinant E. coli Cells and In Vitro Evaluation of Its Potential Prebiotic Effect
Authors: Garcia-Gonzalez Martin , Cervantes Fadia V. , Ipiales Ricardo P. , de la Rubia Angeles , Plou Francisco J. , Fernández-Lobato María
Published: October 2022

In this work, the trisaccharide isomelezitose was overproduced from sucrose using a biocatalyst based on immobilized Escherichia coli cells harbouring the α-glucosidase from the yeast Metschnikowia reukaufii, the best native producer of this sugar described to date. The overall process for isomelezitose production and purification was performed in three simple steps: (i) oligosaccharides synthesis by alginate-entrapped E. coli; (ii) elimination of monosaccharides (glucose and fructose) using alginate-entrapped Komagataella phaffii cells; and (iii) semi-preparative high performance liquid chromatography under isocratic conditions. As result, approximately 2.15 g of isomelezitose (purity exceeding 95%) was obtained from 15 g of sucrose. The potential prebiotic effect of this sugar on probiotic bacteria (Lactobacillus casei, Lactobacillus rhamnosus and Enterococcus faecium) was analysed using in vitro assays for the first time. The growth of all probiotic bacteria cultures supplemented with isomelezitose was significantly improved and was similar to that of cultures supplemented with a commercial mixture of fructo-oligosaccharides. In addition, when isomelezitose was added to the bacteria cultures, the production of organic acids (mainly butyrate) was significantly promoted. Therefore, these results confirm that isomelezitose is a potential novel prebiotic that could be included in healthier foodstuffs designed for human gastrointestinal balance maintenance.
Cheminformatics Study on Structural and Bactericidal Activity of Latest Generation β-Lactams on Widespread Pathogens
Authors: Gherman Ana Maria Raluca , Dina Nicoleta Elena , Chiș Vasile
Published: October 2022

Raman spectra of oxacillin (OXN), carbenicillin (CBC), and azlocillin (AZL) are reported for the first time together with their full assignment of the normal modes, as calculated using Density Functional Theory (DFT) methods with the B3LYP exchange-correlation functional coupled to the 6-31G(d) and 6-311+G(2d,p) basis sets. Molecular docking studies were performed on five penicillins, including OXN, CBC, and AZL. Subsequently, their chemical reactivity and correlated efficiency towards specific pathogenic strains were revealed by combining frontier molecular orbital (FMO) data with molecular electrostatic potential (MEP) surfaces. Their bactericidal activity was tested and confirmed on a couple of species, both Gram-positive and Gram-negative, by using the disk diffusion method. Additionally, a surface-enhanced Raman spectroscopy (SERS)—principal component analysis (PCA)-based resistogram of A. hydrophila is proposed as a clinically relevant insight resulting from the synergistic cheminformatics and vibrational study on CBC and AZL.
Epigenetics and Pregnancy: Conditional Snapshot or Rolling Event
Authors: Andrawus Mariana , Sharvit Lital , Atzmon Gil
Published: October 2022

Epigenetics modification such as DNA methylation can affect maternal health during the gestation period. Furthermore, pregnancy can drive a range of physiological and molecular changes that have the potential to contribute to pathological conditions. Pregnancy-related risk factors include multiple environmental, behavioral, and hereditary factors that can impact maternal DNA methylation with long-lasting consequences. Identification of the epigenetic patterns linked to poor pregnancy outcomes is crucial since changes in DNA methylation patterns can have long-term effects. In this review, we provide an overview of the epigenetic changes that influence pregnancy-related molecular programming such as gestational diabetes, immune response, and pre-eclampsia, in an effort to close the gap in current understanding regarding interactions between the environment, the genetics of the fetus, and the pregnant woman.
A Xanthohumol-Rich Hop Extract Diminishes Endotoxin-Induced Activation of TLR4 Signaling in Human Peripheral Blood Mononuclear Cells: A Study in Healthy Women
Authors: Jung Finn , Staltner Raphaela , Baumann Anja , Burger Katharina , Halilbasic Emina , Hellerbrand Claus , Bergheim Ina
Published: October 2022

Infections with Gram-negative bacteria are still among the leading causes of infection-related deaths. Several studies suggest that the chalcone xanthohumol (XN) found in hop (Humulus lupulus) possesses anti-inflammatory effects. In a single-blinded, placebo controlled randomized cross-over design study we assessed if the oral intake of a single low dose of 0.125 mg of a XN derived through a XN-rich hop extract (75% XN) affects lipopolysaccharide (LPS)-induced immune responses in peripheral blood mononuclear cells (PBMCs) ex vivo in normal weight healthy women (n = 9) (clinicaltrials.gov: NCT04847193) and determined associated molecular mechanisms. LPS-stimulation of PBMCs isolated from participants 1 h after the intake of the placebo for 2 h resulted in a significant induction of pro-inflammatory cytokine release which was significantly attenuated when participants had consumed XN. The XN-dependent attenuation of proinflammatory cytokine release was less pronounced 6 h after the LPS stimulation while the release of sCD14 was significantly reduced at this timepoint. The LPS-dependent activation of hTLR4 transfected HEK293 cells was significantly and dose-dependently suppressed by the XN-rich hop extract which was attenuated when cells were co-challenged with sCD14. Taken together, our results suggest even a one-time intake of low doses of XN consumed in a XN-rich hop extract can suppress LPS-dependent stimulation of PBMCs and that this is related to the interaction of the hop compound with the CD14/TLR4 signaling cascade.
The Mutation Hotspots at UGT1A Locus May Be Associated with Gilbert’s Syndrome Affecting the Taiwanese Population
Authors: Hsu Paul Wei-Che , Liao Po-Cheng , Kao Yu-Hsiang , Lin Xin-Yu , Chien Rong-Nan , Yeh Chau-Ting , Lai Chi-Chun , Shyu Yu-Chiau , Lin Chih-Lang
Published: October 2022

Gilbert’s syndrome is mainly diagnosed through genetic analysis and is primarily detected through a mutation in the promoter region of the UGT1A1 gene. However, most of the research has been conducted on Caucasian populations. In this study, we studied the Han population in Taiwan to investigate the possibility of other mutations that could cause Gilbert’s syndrome. This study comprised a test group of 45 Taiwanese individuals with Gilbert’s syndrome and 180 healthy Taiwanese individuals as a control group. We extracted DNA from the blood samples and then used Axiom Genome-Wide TWB 2.0 array plates for genotyping. Out of 302,771 single nucleotide polymorphisms (SNPs) from 225 subjects, we detected 57 SNPs with the most significant shift in allele frequency; 27 SNPs among them were located in the UGT1A region. Most of the detected SNPs highly correlated with each other and are located near the first exon of UGT1A1, UGT1A3, UGT1A6, and UGT1A7. We used these SNPs as an input for the machine learning algorithms and developed prediction models. Our study reveals a good association between the 27 SNPs detected and Gilbert’s syndrome. Hence, this study provides a reference for diagnosing Gilbert’s syndrome in the Taiwanese population in the future.
Necroptosis: A Pathogenic Negotiator in Human Diseases
Authors: Chaouhan Hitesh Singh , Vinod Ch , Mahapatra Nikita , Yu Shao-Hua , Wang I-Kuan , Chen Kuen-Bao , Yu Tung-Min , Li Chi-Yuan
Published: October 2022

Over the past few decades, mechanisms of programmed cell death have attracted the scientific community because they are involved in diverse human diseases. Initially, apoptosis was considered as a crucial mechanistic pathway for programmed cell death; recently, an alternative regulated mode of cell death was identified, mimicking the features of both apoptosis and necrosis. Several lines of evidence have revealed that dysregulation of necroptosis leads to pathological diseases such as cancer, cardiovascular, lung, renal, hepatic, neurodegenerative, and inflammatory diseases. Regulated forms of necrosis are executed by death receptor ligands through the activation of receptor-interacting protein kinase (RIPK)-1/3 and mixed-lineage kinase domain-like (MLKL), resulting in the formation of a necrosome complex. Many papers based on genetic and pharmacological studies have shown that RIPKs and MLKL are the key regulatory effectors during the progression of multiple pathological diseases. This review focused on illuminating the mechanisms underlying necroptosis, the functions of necroptosis-associated proteins, and their influences on disease progression. We also discuss numerous natural and chemical compounds and novel targeted therapies that elicit beneficial roles of necroptotic cell death in malignant cells to bypass apoptosis and drug resistance and to provide suggestions for further research in this field.
Changes in Polychlorinated Biphenyl Residues in Milk during Lactation: Levels of Contamination, Influencing Factors, and Infant Risk Assessment
Authors: Witczak Agata , Pohoryło Anna , Aftyka Aleksandra , Pokorska-Niewiada Kamila , Witczak Grzegorz
Published: October 2022

Given the importance of breastfeeding infants, the contamination of human milk is a significant public concern. The aim of this study was to assess the contamination of human milk with dioxin-like PCBs (dl-PCBs) and non-dioxin-like PCBs (ndl-PCBs) in relation to the duration of lactation and other influencing factors, especially the frequency of the consumption of selected foods during pregnancy. Based on this, the health risk to infants was assessed and compared to the tolerable daily intake (TDI). PCB determinations were performed using gas chromatography/mass spectrometry. The ∑ndl-PCB content ranged from 0.008 to 0.897 ng/g w.w., at an average of 0.552 ng/g wet weight, which was 55% of the maximum level according to the EU guidelines for foods for infants and young children. The toxic equivalent (TEQ) was in the range of 0.033–5.67 pg-TEQ/g w.w. The content of non-ortho, mono-ortho, and ndl-PCBs in human milk decreased the longer lactation continued. Moreover, when pregnant women smoked tobacco, this correlated significantly with increases in the concentrations of PCB congeners 156, 118, and 189 in human milk. The human milk contents of PCB congeners 77, 81, 186, 118, and 189 were strongly positively correlated with the amount of fish consumed. The content of stable congeners PCB 135 and PCB 153 increased with age.
Genetic and Transcriptome Analysis of Leaf Trichome Development in Chinese Cabbage (Brassica rapa L. subsp. pekinensis) and Molecular Marker Development
Authors: Li Jingjuan , Wang Hongxia , Zhou Dandan , Li Cheng , Ding Qian , Yang Xiaogang , Wang Fengde , Zheng Han , Gao Jianwei
Published: October 2022

Chinese cabbage (Brassica rapa L. subsp. pekinensis) is one of the vegetables with the largest cultivated area in China and has been a great addition to the daily diet of Chinese people. A genetic map has been constructed in our previous study using the F2 population of two inbred lines of Chinese cabbage, namely “G291” (a hairy line) and “ZHB” (a hairless line), based on which a candidate gene related to trichome traits was identified on chromosome A06 with a phenotypic variance of 47%. A molecular marker was found to co-segregate with the trichome traits of the F2 population, which is in the 5′-flanking region of BrGL1, and a corresponding patent has been granted (NO. CN 108545775 B). Transcriptome analysis was carried out on the cotyledon, the first true leaf and the leaf closest to each inflorescence of F2 individuals of “G291 × ZHB” with or without trichomes, respectively. Ten pathways, including 189 DEGs, were identified to be involved in the development of trichomes in Chinese cabbage, which may be specifically related to the development of leaf trichomes. Most of the pathways were related to the biosynthesis of the secondary metabolites, which may help plants to adapt to the ever-changing external environment. DEGs also enriched the “plant-pathogen interaction” pathway, which is consistent with the conclusion that trichomes are related to the disease resistance of plants. Our study provides a basis for future research on the occurrence and development of trichomes in Chinese cabbage.