bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic Ablation of the Inducible Form of Nitric Oxide in Male Mice Disrupts Immature Neuron Survival in the Adult Dentate Gyrus
Authors: Fernandes Gabriel G. , Costa Karla C. M. , Scomparin Davi S. , Freire Juliana B. , Guimarães Francisco S. , Campos Alline C.
Published: December 2021

Inducible nitric oxide synthase (iNOS) is an enzyme upregulated in the brain during neuroimmune stimuli which is associated with an oxidative and pro-inflammatory environment in several brain regions, including the hippocampal formation and the prefrontal cortex. The dentate gyrus of the hippocampal formation is the site of a process known as adult hippocampal neurogenesis (AHN). Although many endogenous and extrinsic factors can modulate AHN, the exact participation of specific proinflammatory mediators such as iNOS in these processes remains to be fully elucidated. Here, we investigated how the total genetic ablation of iNOS impacts the hippocampal neurogenic niche and microglial phenotype and if these changes are correlated to the behavioral alterations observed in iNOS knockout (K.O.) mice submitted or not to the chronic unpredictable stress model (CUS - 21 days protocol). Contrary to our initial hypothesis, at control conditions, iNOS K.O. mice displayed no abnormalities on microglial activation in the dentate gyrus. However, they did exhibit impaired newborn cells and immature neuron survival, which was not affected by CUS. The reduction of AHN in iNOS K.O. mice was accompanied by an increased positive coping response in the tail suspension test and facilitation of anxiety-like behaviors in the novelty suppressed feeding. Next, we investigated whether a pro-neurogenic stimulus would rescue the neurogenic capacity of iNOS K.O. mice by administering in control and CUS groups the antidepressant escitalopram (ESC). The chronic treatment with ESC could not rescue the neurogenic capacity or the behavioral changes observed in iNOS K.O. mice. Besides, in the ventromedial prefrontal (vmPFC) cortex there was no change in the expression or the chronic activation of PV neurons (evaluated by double labeling PV with FOSB) in the prelimbic (PrL) or infralimbic subregions. FOSB expression, however, increased in the PrL of iNOS K.O. mice. Our results suggest that iNOS seems essential for the survival of newborn cells and immature neurons in the hippocampus and seem to partially explain the anxiogenic-like behavior observed in iNOS K.O. mice. On the other hand, the iNOS ablation appears to result in increased activity of the PrL which could explain the antidepressant-like behaviors of iNOS K.O mice.
Crosstalk Between Circulating Follicular T Helper Cells and Regulatory B Cells in Children With Extrinsic Atopic Dermatitis
Authors: Jiang Jinqiu , Yan Shi , Zhou Xiaoying , Zhou Jinghua , Bai Xiaoming , Tan Qi , Xia Yunqiu , Wang Hua , Luo Xiaoyan
Published: December 2021

Atopic dermatitis (AD) in early childhood is often the initial manifestation of allergic disease associated with high IgE. Accumulating evidences show that follicular helper T (Tfh) cells play a critical role in promoting B cell differentiation and IgE production, human regulatory B (Breg) cells participate in immunomodulatory processes and inhibition of allergic inflammation. However, the roles and interactions between IL-10-producing Breg cells and Tfh cells in childhood AD are unclear. In this study, we found that the percentage of CD19+IL-10+ Breg cells in children with extrinsic AD was significantly lower than that in age-matched healthy controls, and that it correlated negatively with enhanced CD4+CXCR5+PD-1+ICOS+ circulating Tfh cell responses and increased disease activity; however, there was no significant correlation with serum total IgE levels. A co-culture system revealed that Breg cells from patients with extrinsic AD cannot effectively inhibit differentiation of Tfh cells in an IL-10 dependent manner. Abnormal pSTAT3 signaling induced via Toll-like receptors (TLR), but not the B-cell receptor (BCR) signaling, might contribute to the defect of Breg cells in AD. Taken together, these observations demonstrate an important role for IL-10-producing Breg cells in inhibiting Tfh cell differentiation, and suggest that they may participate in the pathogenesis of AD.
Expression Dynamics of the O-Glycosylated Proteins Recognized by Amaranthus leucocarpus Lectin in T Lymphocytes and Its Relationship With Moesin as an Alternative Mechanism of Cell Activation
Authors: Gómez-Henao Wilton , Saavedra Rafael , Chávez-Sánchez Francisco Raúl , Lascurain Ricardo , Zenteno Edgar , Tenorio Eda Patricia
Published: December 2021

T lymphocyte activation begins with antigen/MHC recognition by the TCR/CD3 complex followed by a costimulatory signal provided by CD28. The search for novel costimulatory molecules has been extensive due to their potential use as immunotherapeutic targets. Although some molecules have been identified, they are unable to provide sustainable signaling to allow for proper T cell activation and proliferation. It has been shown that the Amaranthus leucocarpus lectin (ALL) can be used as an in vitro costimulator of CD4+ lymphocytes in the presence of anti-CD3 mAb; this lectin specifically recognizes O-glycans of the Galβ1-3GalNAc-O-Ser/Thr type, including a 70-kDa moesin-like protein that has been suggested as the costimulatory molecule. However, the identity of this molecule has not been confirmed and such costimulation has not been analyzed in CD8+ lymphocytes. We show herein that the expression kinetics of the glycoproteins recognized by ALL (gpALL) is different in CD4+ and CD8+ T cells, unlike moesin expression. Results from IP experiments demonstrate that the previously described 70-kDa moesin-like protein is an O-glycosylated form of moesin (O-moesin) and that in vitro stimulation with anti-CD3 and anti-moesin mAb induces expression of the activation molecules CD69 and CD25, proliferation and IL-2 production as efficiently as cells costimulated with ALL or anti-CD28. Overall, our results demonstrate that O-moesin is expressed in CD4+ and CD8+ T lymphocytes and that moesin provides a new costimulatory activation signal in both T cell subsets.
A Genome-Wide CRISPR/Cas9-Based Screen Identifies Heparan Sulfate Proteoglycans as Ligands of Killer-Cell Immunoglobulin-Like Receptors
Authors: Klein Klara , Hölzemer Angelique , Wang Tim , Kim Tae-Eun , Dugan Haley L. , Jost Stephanie , Altfeld Marcus , Garcia-Beltran Wilfredo F.
Published: December 2021

While human leukocyte antigen (HLA) and HLA-like proteins comprise an overwhelming majority of known ligands for NK-cell receptors, the interactions of NK-cell receptors with non-conventional ligands, particularly carbohydrate antigens, is less well described. We previously found through a bead-based HLA screen that KIR3DS1, a formerly orphan member of the killer-cell immunoglobulin-like receptor (KIR) family, binds to HLA-F. In this study, we assessed the ligand binding profile of KIR3DS1 to cell lines using Fc fusion constructs, and discovered that KIR3DS1-Fc exhibited binding to several human cell lines including ones devoid of HLA. To identify these non-HLA ligands, we developed a magnetic enrichment-based genome-wide CRISPR/Cas9 knock-out screen approach, and identified enzymes involved in the biosynthesis of heparan sulfate as crucial for the binding of KIR3DS1-Fc to K562 cells. This interaction between KIR3DS1 and heparan sulfate was confirmed via surface plasmon resonance, and removal of heparan sulfate proteoglycans from cell surfaces abolished KIR3DS1-Fc binding. Testing of additional KIR-Fc constructs demonstrated that KIR family members containing a D0 domain (KIR3DS1, KIR3DL1, KIR3DL2, KIR2DL4, and KIR2DL5) bound to heparan sulfate, while those without a D0 domain (KIR2DL1, KIR2DL2, KIR2DL3, and KIR2DS4) did not. Overall, this study demonstrates the use of a genome-wide CRISPR/Cas9 knock-out strategy to unbiasedly identify unconventional ligands of NK-cell receptors. Furthermore, we uncover a previously underrecognized binding of various activating and inhibitory KIRs to heparan sulfate proteoglycans that may play a role in NK-cell receptor signaling and target-cell recognition.
Immune Evaluation of Recombinant Lactobacillus plantarum With Surface Display of HA1-DCpep in Mice
Authors: Niu Hui , Xing Jun-Hong , Zou Bo-Shi , Shi Chun-Wei , Huang Hai-Bin , Jiang Yan-Long , Wang Jian-Zhong , Cao Xin , Wang Nan , Zeng Yan , Yang Wen-Tao , Yang Gui-Lian , Wang Chun-Feng
Published: December 2021

Avian influenza viruses can be efficiently transmitted through mucous membranes, and conventional vaccines are not effective in protecting against mucosal infection by influenza viruses. To induce multiple immune responses in an organism, we constructed a recombinant Lactobacillus plantarum expressing the influenza virus antigen HA1 with the adjuvant dendritic cell-targeting peptide (DCpep). The recombinant L. plantarum strains NC8Δ-pWCF-HA1 and NC8Δ-pWCF-HA1-DCpep were used to immunize mice via oral administration, and the humoral, cellular and mucosal immune responses were evaluated. In addition, the serum levels of specific antibodies and hemagglutination inhibition (HI) levels were also measured. Our results showed that recombinant L. plantarum activated dendritic cells in Peyer’s patches (PPs), increased the numbers of CD4+IFN-γ+ and CD8+IFN-γ+ cells in the spleen and mesenteric lymph nodes (MLNs), and affected the ability of CD4+ and CD8+ cells to proliferate in the spleen and MLNs. Additionally, recombinant L. plantarum increased the number of B220+IgA+ cells in PPs and the level of IgA in the lungs and different intestinal segments. In addition, specific IgG, IgG1 and IgG2a antibodies were induced at high levels in the mice serum, specific IgA antibodies were induced at high levels in the mice feces, and HI potency was significantly increased. Thus, the recombinant L. plantarum strains NC8Δ-pWCF-HA1 and NC8Δ-pWCF-HA1-DCpep have potential as vaccine candidates for avian influenza virus.
Application of a Multidisciplinary Weight of Evidence Approach as a Tool for Monitoring the Ecological Risk of Dredging Activities
Authors: d'Errico Giuseppe , Nardi Alessandro , Benedetti Maura , Mezzelani Marica , Fattorini Daniele , Di Carlo Marta , Pittura Lucia , Giuliani Maria Elisa , Macchia Simona , Vitiello Valentina , Sartori Davide , Scuderi Alice , Morroni Lorenzo , Chiaretti Gianluca , Gorbi Stefania , Pellegrini David , Regoli Francesco
Published: December 2021

The use of multidisciplinary investigations for the evaluation of aquatic ecosystems status is recommended by the European Directives, but it is still a challenging practice. In this study, we apply a quantitative weight of evidence (WOE) approach (Sediqualsoft) for the integration of extensive data obtained from different typologies of investigations, obtained over a 4-year monitoring study of dredging activities in the harbor of Leghorn (Italy). During different phases of such operations, selected sites have been characterized in terms of levels of trace metals and polycyclic aromatic hydrocarbons in sediments, bioaccumulation of contaminants, and a wide battery of biomarkers in transplanted mussels, ecotoxicological effects of sediments through a battery of bioassays (algal growth inhibition, bioluminescence inhibition, and embryotoxicity tests), and the status of benthic communities. Each typology of data, line of evidence (LOE), has been initially elaborated through dedicated logical flowcharts and algorithms providing specific hazard indices, followed by their overall integration based on different weights assigned to each LOE. This approach allowed to summarize more than 10,000 results, reaching robust conclusions on environmental impact during various phases of dredging operations. This approach was confirmed as a useful tool for monitoring the risk, supporting a “site-oriented” decision making process by providing stakeholders simple interpretation of complex data.
Influences of Salinity on Embryonic and Larval Development of Striped Catfish Pangasianodon hypophthalmus
Authors: Hossain Farzana , Islam S. M. Majharul , Ashaf-Ud-Doulah Mohammad , Ali Md Shaha , Islam Md Sadiqul , Brown Christopher , Shahjahan Md
Published: December 2021

Salinity intrusion in coastal areas due to climate change is alarming. In this study, the effects of salinity on embryonic and larval development of striped catfish (Pangasianodon hypophthalmus) were studied experimentally. Embryos and larvae were exposed to seven salinity treatments (0, 2, 4, 6, 8, 10, and 12 ppt), each with three replications. Considerable survivability of embryos was recorded up to 6 ppt salinity. Mortality of embryos significantly increased at 8 and 10 ppt salinity, and 100% mortality was displayed within 12 h of exposure at 12 ppt salinity. The rate of hatching was significantly reduced at 8 and 10 ppt salinity. The 24 h lethal concentration (LC50) value of salinity for embryo was 11.24 ppt. Different types of deformities, such as undeveloped yolk sac, elongated gastrula yolk sac, and yolk sac bud, were highest at 10 ppt salinity. Similar to the embryo, considerable survivability of larvae was recorded up to 6 ppt salinity, and 100% mortalities were found within 24 h of exposure at 12 ppt salinity. The 24 and 48 h LC50 values of salinity for larvae were 10.63 and 8.48 ppt, respectively. Several types of deformities, including yolk sac ulceration, spine scoliosis, tail bent, yolk sac edema, and compromised swim bladder inflation, were highest at 10 ppt salinity after 48 h of exposure. Within 24 h of exposure, about 80% yolk sac of the larvae was absorbed at 8 and 10 ppt salinity, while 30%–50% yolk sac was absorbed at 0–6 ppt salinity. Growth rates in terms of length and weight were higher at 0, 2, and 4 ppt salinity and moderate at 6 and 8 ppt salinity. Overall, the current findings define the limits to optimize hatchery procedures for the culture of this species in low saline brackish water.
Non-invasive Early Prediction of Septic Acute Kidney Injury by Doppler-Based Renal Resistive Indexes Combined With Echocardiographic Parameters: An Experimental Study
Authors: Zhang Ying , Zhu Jianing , Zhang Chuyue , Xiao Jing , Liu Chao , Wang Shuo , Zhao Ping , Zhu Yaqiong , Wang Li , Li Qiuyang , Luo Yukun
Published: December 2021

Non-invasive early prediction of septic acute kidney injury (S-AKI) is still urgent and challenging. Increased Doppler-based renal resistive index (RRI) has been shown to be associated with S-AKI, but its clinical use is limited, which may be explained by the complex effects of systemic circulation. Echocardiogram allows non-invasive assessment of systemic circulation, which may provide an effective supplement to RRI. To find the value of RRI combined with echocardiographic parameters in the non-invasive early prediction of S-AKI, we designed this experiment with repeated measurements of ultrasonographic parameters in the early stage of sepsis (3, 6, 12, and 24 h) in cecum ligation and puncture (CLP) rats (divided into AKI and non-AKI groups at 24 h based on serum creatinine), with sham-operated group serving as controls. Our results found that RRI alone could not effectively predict S-AKI, but when combined with echocardiographic parameters (heart rate, left ventricular end-diastolic internal diameter, and left ventricular end-systolic internal diameter), the predictive value was significantly improved, especially in the early stage of sepsis (3 h, AUC: 0.948, 95% CI 0.839–0.992, P < 0.001), and far earlier than the conventional renal function indicators (serum creatinine and blood urea nitrogen), which only significantly elevated at 24 h. Our method showed novel advances and potential in the early detection of S-AKI.
Quantitative Brain Positron Emission Tomography in Female 5XFAD Alzheimer Mice: Pathological Features and Sex-Specific Alterations
Authors: Bouter Caroline , Irwin Caroline , Franke Timon N. , Beindorff Nicola , Bouter Yvonne
Published: December 2021

Successful back-translating clinical biomarkers and molecular imaging methods of Alzheimer's disease (AD), including positron emission tomography (PET), are very valuable for the evaluation of new therapeutic strategies and increase the quality of preclinical studies. 18F-Fluorodeoxyglucose (FDG)–PET and 18F-Florbetaben–PET are clinically established biomarkers capturing two key pathological features of AD. However, the suitability of 18F-FDG– and amyloid–PET in the widely used 5XFAD mouse model of AD is still unclear. Furthermore, only data on male 5XFAD mice have been published so far, whereas studies in female mice and possible sex differences in 18F-FDG and 18F-Florbetaben uptake are missing. The aim of this study was to evaluate the suitability of 18F-FDG– and 18F-Florbetaben–PET in 7-month-old female 5XFAD and to assess possible sex differences between male and female 5XFAD mice. We could demonstrate that female 5XFAD mice showed a significant reduction in brain glucose metabolism and increased cerebral amyloid deposition compared with wild type animals, in accordance with the pathology seen in AD patients. Furthermore, we showed for the first time that the hypometabolism in 5XFAD mice is gender-dependent and more pronounced in female mice. Therefore, these results support the feasibility of small animal PET imaging with 18F-FDG- and 18F-Florbetaben in 5XFAD mice in both, male and female animals. Moreover, our findings highlight the need to account for sex differences in studies working with 5XFAD mice.
Assessment of Prognostic Value of Aspartate Aminotransferase-to-Platelet Ratio Index in Patients With Hepatocellular Carcinoma: Meta-Analysis of 28 Cohort Studies
Authors: Zhang XinYue , Svn Zhen , Liv MengSi , Liu MengNan , Zhang YiHan , Sun Qin
Published: December 2021

Background: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors globally; it is valuable to predict its prognosis after treatment. Aspartate aminotransferase-to-platelet index (APRI), a non-invasive biomarker consists of two routine test parameters easily available in all the patients. Our study aimed to investigate whether APRI can serve as an independent prognostic marker in the patients with HCC.

Methods: We extensively searched PubMed, Embase, and Web of Science databases on June 20, 2021 to determine all relevant literature. The studies that explored the association between the APRI levels and prognosis of patients with HCC and reported risk estimate data were included. The Newcastle-Ottawa Scale was used to assess the quality of the included studies.

Results: A total of 1,097 articles were initially identified, of which 28 studies involving 11,041 patients met the eligibility criteria for the meta-analysis. The pooled hazard ratios (HRs) for overall survival (OS) and disease-free survival (DFS) were 1.77 (95% CI: 1.53–2.05, P < 0.001) and 1.59 (95% CI: 1.47–1.71, P < 0.001), respectively, suggesting a significant correlation between the increased APRI levels and poor prognosis in the patients with HCC. In the subgroup analyses, statistical significance of the correlation disappeared in the Korean and Japanese population and in the patients undergoing transarterial chemoembolization (TACE). Of note, the current results may be overestimated due to publication bias, but the conclusion remained unchanged when the bias was adjusted.

Conclusion: High APRI levels are associated with poor OS and DFS in the patients with HCC. In most cases, pretreatment APRI can be used as an independent prognostic factor, but it is necessary to incorporate other predictive prognostic systems to ensure accuracy. Further studies are needed to determine the specific beneficiary population and the optimal cutoff value.
The Prevalence and Determinants of Fusidic Acid Resistance Among Methicillin-Resistant Staphylococcus aureus Clinical Isolates in China
Authors: Zhao Huilin , Wang Xinyi , Wang Bingjie , Xu Yanlei , Rao Lulin , Wan Baoshan , Guo Yinjuan , Wu Xiaocui , Yu Jingyi , Chen Liang , Li Meilan , Yu Fangyou
Published: December 2021

The significant increase in resistance of methicillin-resistant Staphylococcus aureus (MRSA) to fusidic acid (FA) is a worrying public concern. However, the data on the prevalence of FA-resistant MRSA isolates in China is still limited. This study aims to investigate the prevalence of FA resistance and resistance determinants among MRSA isolates from six tertiary hospitals in different regions of China between 2016 and 2020. The antimicrobial susceptibility of MRSA isolates was performed by disk diffusion test and broth microdilution method. Whole-genome sequencing was conducted to evaluate the determinants of FA resistance and molecular characterization of FA-resistant MRSA isolates. In this study, a total of 74 (74/457, 16.2%) isolates were identified to be FA-resistant among 457 non-duplicate MRSA isolates. The prevalence of 74 FA-resistant isolates was as follows: Hubei (28/70, 40%), Shanghai (18/84, 21.4%), Jiangxi (7/58, 12.1%), Inner Mongolia Autonomous Region (6/38, 15.8%), Guangdong (12/112, 10.7%), and Sichuan (3/95, 3.2%). The mutations in fusA were present in 79.7% (59/74) of FA-resistant MRSA isolates, with 54 (54/74, 73%) having L461K mutation and conferring high-level resistance [Minimum Inhibitory Concentration (MIC)>128 μg/ml]. Acquired gene, fusB, with low-level resistance (MIC <16 μg/ml) was found in 20.3% (15/74) FA-resistant MRSA isolates. ST5-MRSA-II-t2460 was the most prevalence clone with high-level resistance, accounting for 51.4% (38/74), which was distributed in Hubei (24/28, 85.7%), Inner Mongolia Autonomous Region (4/6, 66.7%), Shanghai (7/18, 38.9%), and Guangdong (3/12, 25%). ST630-t4549 MRSA isolates with low-level resistance were the most common in Jiangxi (3/7, 42.9%) and Sichuan (2/3, 66.7%). In brief, the prevalence of FA resistance among MRSA isolates in China was relatively high with geographic differences. High-level FA resistance was associated mostly with fusA mutations, especially the L461K mutation, whereas fusB usually conferred the low-level resistance to FA. The spread of ST5-MRSA-II-t2460 clone with high-level resistance to FA contributed greatly to the increase of FA-resistant MRSA isolates in most regions, especially in Hubei.
Estimated Annual Economic Burden of Dry Eye Disease Based on a Multi-Center Analysis in China: A Retrospective Study
Authors: Yang Wanju , Luo Yanzhu , Wu Shangcao , Niu Xiaoxia , Yan Yanshuang , Qiao Chen , Ming Wei , Zhang Ying , Wang Haoyu , Chen Dan , Qi Mengying , Ke Lan , Wang Ying , Li Liping , Li Shaowei , Zeng Qingyan
Published: December 2021

Purpose: To conduct a multi-center analysis and assess the economic burden due to dry eye disease (DED) in China.

Design: A retrospective and cross-sectional study.

Methods: Patients (n = 598) with diagnosed DED were recruited from 3 eye centers (in central, southeast, and northeast China) from 1 January 2018 to 31 December 2018. Data were collected regarding the examination, pharmacological therapy, and non-pharmacological therapy fees. Sub-group analyses were stratified by eye center, DED severity, types of DED, number of visits to physicians, and residential area. A logistic regression analysis was conducted to investigate the variables influencing total costs.

Results: The per capita costs devoted to DED at the 3 centers were 422.6, 391.3, and 265.4 USD, respectively. The costs of non-pharmacological therapy accounted the largest part in three centers (75.6, 76.4, 76.5%, respectively). Patients with severe DED sustained the largest economic burden. Patients with mixed type of DED spent the most comparing to patients with either evaporative or aqueous-deficient types of DED. Patients spent more during the first visit compared with subsequent visits. Patients living in urban areas spent significantly more than did those living in rural areas (P = 0.001). The logistics regression analysis showed that total costs were significantly influenced by DED severity, number of visits to physicians, and area of residence (beta = 2.83, 0.83, 1.48; P < 0.0001).

Conclusions: DED is a chronic ocular disease that timely non-cost counseling, early diagnosis, and efficacious treatment can reduce its economic burden on patients and the society.
Biofloc Microbiome With Bioremediation and Health Benefits
Authors: Kumar Vikash , Roy Suvra , Behera Bijay Kumar , Swain Himanshu Sekhar , Das Basanta Kumar
Published: December 2021

The biofloc system has recently attracted great attention as a cost-effective, sustainable, and environmentally friendly technology and expected to contribute toward human food security (Zero Hunger SDG 2). It is also expected that this endeavor can be adopted widely because of its characteristics of zero water exchange and reduced artificial feeding features. In the biofloc system, the flocs which are generally formed by aggregation of heterotrophic microorganisms, serve as natural bioremediation candidates. These microbes effectively maintain water quality by utilizing the nutrient wastes, mostly originated from digested, unconsumed, and metabolic processes of feed. Additionally, the flocs are important sources of nutrients, mainly a protein source, and when these are consumed by aquaculture animals they improve the growth performance, immunity, and disease tolerance of host against pathogenic microbial infection. Here in this review, we focus on recent advances that could provide a mechanistic insight on how the microbial community developed in the biofloc system helps in the bioremediation process and enhances the overall health of the host. We have also tried to address the possible role of these microbial communities against growth and virulence of pathogenic microbes.
Essential Oil Nanoemulsion as Eco-Friendly and Safe Preservative: Bioefficacy Against Microbial Food Deterioration and Toxin Secretion, Mode of Action, and Future Opportunities
Authors: Maurya Akash , Singh Vipin Kumar , Das Somenath , Prasad Jitendra , Kedia Akash , Upadhyay Neha , Dubey Nawal Kishore , Dwivedy Abhishek Kumar
Published: December 2021

Microbes are the biggest shareholder for the quantitative and qualitative deterioration of food commodities at different stages of production, transportation, and storage, along with the secretion of toxic secondary metabolites. Indiscriminate application of synthetic preservatives may develop resistance in microbial strains and associated complications in human health with broad-spectrum environmental non-sustainability. The application of essential oils (EOs) as a natural antimicrobial and their efficacy for the preservation of foods has been of present interest and growing consumer demand in the current generation. However, the loss in bioactivity of EOs from fluctuating environmental conditions is a major limitation during their practical application, which could be overcome by encapsulating them in a suitable biodegradable and biocompatible polymer matrix with enhancement to their efficacy and stability. Among different nanoencapsulated systems, nanoemulsions effectively contribute to the practical applications of EOs by expanding their dispersibility and foster their controlled delivery in food systems. In line with the above background, this review aims to present the practical application of nanoemulsions (a) by addressing their direct and indirect (EO nanoemulsion coating leading to active packaging) consistent support in a real food system, (b) biochemical actions related to antimicrobial mechanisms, (c) effectiveness of nanoemulsion as bio-nanosensor with large scale practical applicability, (d) critical evaluation of toxicity, safety, and regulatory issues, and (e) market demand of nanoemulsion in pharmaceuticals and nutraceuticals along with the current challenges and future opportunities.
Genetic Diversity and Natural Selection of Plasmodium vivax Duffy Binding Protein-II From China-Myanmar Border of Yunnan Province, China
Authors: Shi Tian-Qi , Shen Hai-Mo , Chen Shen-Bo , Kassegne Kokouvi , Cui Yan-Bing , Xu Bin , Chen Jun-Hu , Zheng Bin , Wang Yue
Published: December 2021

Malaria incidence has declined dramatically over the past decade and China was certified malaria-free in 2021. However, the presence of malaria in border areas and the importation of cases of malaria parasites are major challenges for the consolidation of the achievements made by China. Plasmodium vivax Duffy binding protein (PvDBP) performs a significant role in erythrocyte invasion, and is considered a promising P. vivax vaccine. However, the highly polymorphic region of PvDBP (PvDBP-II) impedes the development of blood-stage vaccine against P. vivax. In this study, we investigated the genetic diversity and natural selection of PvDBP-II among 124 P. vivax isolates collected from the China-Myanmar border (CMB) in Yunnan Province, China, during 2009–2011. To compare genetic diversity, natural selection, and population structure with CMB isolates, 85 pvdbp-II sequences of eastern Myanmar isolates were obtained from GenBank. In addition, global sequences of pvdbp-II were retrieved from GenBank to establish genetic differentiation relationships and networks with the CMB isolates. In total, 22 single nucleotide polymorphisms reflected in 20 non-synonymous and two synonymous mutations were identified. The overall nucleotide diversity of PvDBP-II from the 124 CMB isolates was 0.0059 with 21 haplotypes identified (Hd = 0.91). The high ratio of non-synonymous to synonymous mutations suggests that PvDBP-II had evolved under positive selection. Population structure analysis of the CMB and eastern Myanmar isolates were optimally grouped into five sub-populations (K = 5). Polymorphisms of PvDBP-II display that CMB isolates were genetically diverse. Mutation, recombination, and positive selection promote polymorphism of PvDBP-II of P. vivax population. Although low-level genetic differentiation in eastern Myanmar was identified along with the more effective malaria control measures, the complexity of population structure in malaria parasites has maintained. In conclusion, findings from this study advance knowledge of the understanding of the dynamic of P. vivax population, which will contribute to guiding the rational design of a PvDBP-II based vaccine.
Manganese Stress Adaptation Mechanisms of Bacillus safensis Strain ST7 From Mine Soil
Authors: Ran Xueqin , Zhu Zhongmei , Long Hong , Tian Qun , You Longjiang , Wu Xingdiao , Liu Qin , Huang Shihui , Li Sheng , Niu Xi , Wang Jiafu
Published: December 2021

The mechanism of bacterial adaption to manganese-polluted environments was explored using 50 manganese-tolerant strains of bacteria isolated from soil of the largest manganese mine in China. Efficiency of manganese removal by the isolated strains was investigated using atomic absorption spectrophotometry. Bacillus safensis strain ST7 was the most effective manganese-oxidizing bacteria among the tested isolates, achieving up to 82% removal at a Mn(II) concentration of 2,200 mg/L. Bacteria-mediated manganese oxide precipitates and high motility were observed, and the growth of strain ST7 was inhibited while its biofilm formation was promoted by the presence of Mn(II). In addition, strain ST7 could grow in the presence of high concentrations of Al(III), Cr(VI), and Fe(III). Genome-wide analysis of the gene expression profile of strain ST7 using the RNA-seq method revealed that 2,580 genes were differently expressed under Mn(II) exposure, and there were more downregulated genes (n = 2,021) than upregulated genes (n = 559) induced by Mn stress. KAAS analysis indicated that these differently expressed genes were mainly enriched in material metabolisms, cellular processes, organism systems, and genetic and environmental information processing pathways. A total of twenty-six genes from the transcriptome of strain ST7 were involved in lignocellulosic degradation. Furthermore, after 15 genes were knocked out by homologous recombination technology, it was observed that the transporters, multicopper oxidase, and proteins involved in sporulation and flagellogenesis contributed to the removal of Mn(II) in strain ST7. In summary, B. safensis ST7 adapted to Mn exposure by changing its metabolism, upregulating cation transporters, inhibiting sporulation and flagellogenesis, and activating an alternative stress-related sigB pathway. This bacterial strain could potentially be used to restore soil polluted by multiple heavy metals and is a candidate to support the consolidated bioprocessing community.
Ancient Metabolisms of a Thermophilic Subseafloor Bacterium
Authors: Smith Amy R. , Mueller Ryan , Fisk Martin R. , Colwell Frederick S.
Published: December 2021

The ancient origins of metabolism may be rooted deep in oceanic crust, and these early metabolisms may have persisted in the habitable thermal anoxic aquifer where conditions remain similar to those when they first appeared. The Wood–Ljungdahl pathway for acetogenesis is a key early biosynthetic pathway with the potential to influence ocean chemistry and productivity, but its contemporary role in oceanic crust is not well established. Here, we describe the genome of a novel acetogen from a thermal suboceanic aquifer olivine biofilm in the basaltic crust of the Juan de Fuca Ridge (JdFR) whose genome suggests it may utilize an ancient chemosynthetic lifestyle. This organism encodes the genes for the complete canonical Wood–Ljungdahl pathway, but is potentially unable to use sulfate and certain organic carbon sources such as lipids and carbohydrates to supplement its energy requirements, unlike other known acetogens. Instead, this organism may use peptides and amino acids for energy or as organic carbon sources. Additionally, genes involved in surface adhesion, the import of metallic cations found in Fe-bearing minerals, and use of molecular hydrogen, a product of serpentinization reactions between water and olivine, are prevalent within the genome. These adaptations are likely a reflection of local environmental micro-niches, where cells are adapted to life in biofilms using ancient chemosynthetic metabolisms dependent on H2 and iron minerals. Since this organism is phylogenetically distinct from a related acetogenic group of Clostridiales, we propose it as a new species, Candidatus Acetocimmeria pyornia.
Development of an In-House Rapid Antimicrobial Susceptibility Testing Protocol for Positive Blood Culture and Its Implementation in Routine Microbiology Laboratories
Authors: Cao Min , Huang Lin , Hu Yanyan , Fang Yinfei , Zhang Rong , Chen Gongxiang
Published: December 2021

Bloodstream infections (BSI) are associated with high morbidity and mortality and remain a leading cause of death. Blood culture (BC) including the identification and the antimicrobial susceptibility testing of the causative microorganisms should be performed as soon as possible. In this study, we developed an in-house rapid antimicrobial susceptibility testing (rAST) protocol for positive BC. First, the rAST was performed in the simulated positive BC of standard strains (Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, and Pseudomonas aeruginosa ATCC 27853) at three different times to assess the reproducibility and operability by dispensing four drops of BC broth onto a Mueller–Hinton agar plate after a positive signal. Furthermore, the rAST was performed in clinical positive BCs. The results of rAST at 4, 6, 8, and 18 h of incubation were compared with results of the standard 16- to 20-h disk diffusion method, and the preliminary breakpoints of the rAST method were established according to the inhibition diameter of sensitive strains and resistant strains. Finally, the rAST was performed in the simulated positive BC of clinical strains to evaluate the availability of the preliminary breakpoints. The rAST results of standard strains were distributed evenly at three different times. Among the 202 clinical strains used to establish the preliminary breakpoints, the number of zone diameters that could be read and interpreted (60, 87, 98, and 100%) increased with incubation time (4, 6, 8, and 18 h), and the categorical agreement was acceptable, with total error rates of 3.0, 2.3, 2.1, and 1.3% at 4, 6, 8, and 18 h of incubation, respectively. In conclusion, the in-house rAST protocol for positive BC can be implemented in routine laboratories. It provides reliable antimicrobial susceptibility testing results for BSI pathogens after 4–6 h of incubation.
Suppression and Activation of Intracellular Immune Response in Initial Severe Acute Respiratory Syndrome Coronavirus 2 Infection
Authors: Jia Lijia , Chen Zhen , Zhang Yecheng , Ma Li , Wang Liying , Hu Xiao , Liu Haizhou , Chen Jianjun , Liu Di , Guan Wuxiang
Published: December 2021

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is currently the most important emerging pathogen worldwide, but its early transcriptional dynamics and host immune response remain unclear. Herein, the expression profiles of viral interactions with different types of hosts were comprehensively dissected to shed light on the early infection strategy of SARS-CoV-2 and the host immune response against infection. SARS-CoV-2 was found to exhibit a two-stage transcriptional strategy within the first 24 h of infection, comprising a lag phase that ends with the virus being paused and a log phase that starts when the viral load increases rapidly. Interestingly, the host innate immune response was found not to be activated (latent period) until the virus entered the log stage. Noteworthy, when intracellular immunity is suppressed, SARS-CoV-2 shows a correlation with dysregulation of metal ion homeostasis. Herein, the inhibitory activity of copper ions against SARS-CoV-2 was further validated in in vitro experiments. Coronavirus disease 2019-related genes (including CD38, PTX3, and TCN1) were also identified, which may serve as candidate host-restricted factors for interventional therapy. Collectively, these results confirm that the two-stage strategy of SARS-CoV-2 effectively aids its survival in early infection by regulating the host intracellular immunity, highlighting the key role of interferon in viral infection and potential therapeutic candidates for further investigations on antiviral strategies.
Potential Use of Microbial Enzymes for the Conversion of Plastic Waste Into Value-Added Products: A Viable Solution
Authors: Tamoor Muhammad , Samak Nadia A. , Jia Yunpu , Mushtaq Muhammad Umar , Sher Hassan , Bibi Maryam , Xing Jianmin
Published: December 2021

The widespread use of commercial polymers composed of a mixture of polylactic acid and polyethene terephthalate (PLA-PET) in bottles and other packaging materials has caused a massive environmental crisis. The valorization of these contaminants via cost-effective technologies is urgently needed to achieve a circular economy. The enzymatic hydrolysis of PLA-PET contaminants plays a vital role in environmentally friendly strategies for plastic waste recycling and degradation. In this review, the potential roles of microbial enzymes for solving this critical problem are highlighted. Various enzymes involved in PLA-PET recycling and bioconversion, such as PETase and MHETase produced by Ideonella sakaiensis; esterases produced by Bacillus and Nocardia; lipases produced by Thermomyces lanuginosus, Candida antarctica, Triticum aestivum, and Burkholderia spp.; and leaf-branch compost cutinases are critically discussed. Strategies for the utilization of PLA-PET’s carbon content as C1 building blocks were investigated for the production of new plastic monomers and different value-added products, such as cyclic acetals, 1,3-propanediol, and vanillin. The bioconversion of PET-PLA degradation monomers to polyhydroxyalkanoate biopolymers by Pseudomonas and Halomonas strains was addressed in detail. Different solutions to the production of biodegradable plastics from food waste, agricultural residues, and polyhydroxybutyrate (PHB)-accumulating bacteria were discussed. Fuel oil production via PLA-PET thermal pyrolysis and possible hybrid integration techniques for the incorporation of thermostable plastic degradation enzymes for the conversion into fuel oil is explained in detail.
Development of a Novel Avian Vaccine Vector Derived From the Emerging Fowl Adenovirus 4
Authors: Pan Qing , Zhang Yu , Liu Aijing , Cui Hongyu , Gao Yulong , Qi Xiaole , Liu Changjun , Zhang Yanping , Li Kai , Gao Li , Wang Xiaomei
Published: December 2021

Severe hepatitis-hydropericardium syndrome (HHS) associated with a novel viral genotype, fowl adenovirus 4 (FAdV-4), has emerged and widely spread in China since 2015, causing severe economic losses to the poultry industry. We previously reported that the hexon gene is responsible for pathogenicity and obtained a non-pathogenic hexon-replacement rHN20 strain; however, the lack of information about the non-essential regions for virus replication limits the development of a FAdV-4 vector. This study first established an enhanced green fluorescent protein (EGFP)-indicator virus based on the FAdV-4 reverse genetic technique, effective for batch operations in the virus genome. Based on this, 10 open reading frames (ORFs) at the left end and 13 ORFs at the right end of the novel FAdV-4 genome were deleted separately and identified as non-essential genes for viral replication, providing preliminary insertion sites for foreign genes. To further improve its feasibility as a vaccine vector, seven combinations of ORFs were successfully replaced with EGFP without affecting the immunogenicity of the vector backbone. Finally, a recombinant rHN20-vvIBDV-VP2 strain, expressing the VP2 protein of very virulent infectious bursa disease virus (vvIBDV), was rescued and showed complete protection against FAdV-4 and vvIBDV. Thus, the novel FAdV-4 vector could provide sufficient protection for HHS and efficient exogenous gene delivery. Overall, our findings systemically identified 23 non-essential ORFs for FAdV-4 replication and seven foreign gene insertion regions, providing valuable information for an in-depth understanding of the novel FAdV-4 pathogenesis and development of multivalent vaccines.
In-situ and Real-Time Monitoring of the Interaction Between Lysins and Staphylococcus aureus Biofilm by Surface Plasmon Resonance
Authors: Hong Wei , Nyaruaba Raphael , Li Xiaohong , Liu Huan , Yang Hang , Wei Hongping
Published: December 2021

Staphylococcus aureus can produce a multilayered biofilm embedded in extracellular polymeric matrix. This biofilm is difficult to remove, insensitive to antibiotics, easy to develop drug-resistant strains and causes enormous problems to environments and health. Phage lysin which commonly consists of a catalytic domain (CD) and a cell-wall binding domain (CBD) is a powerful weapon against bacterial biofilm. However, the real-time interaction between lysin and S. aureus biofilm is still not fully understood. In this study, we monitored the interactions of three lysins (ClyF, ClyC, PlySs2) against culture-on-chip S. aureus biofilm, in real-time, based on surface plasmon resonance (SPR). A typical SPR response curve showed that the lysins bound to the biofilm rapidly and the biofilm destruction started at a longer time. By using 1:1 binding model analysis, affinity constants (KD) for ClyF, ClyC, and PlySs2 were found to be 3.18 ± 0.127 μM, 1.12 ± 0.026 μM, and 15.5 ± 0.514 μM, respectively. The fact that ClyF and PlySs2 shared the same CBD but showed different affinity to S. aureus biofilm suggested that, not only CBD, but also CD affects the binding activity of the entire lysin. The SPR platform can be applied to improve our understanding on the complex interactions between lysins and bacterial biofilm including association (adsorption) and disassociation (destruction).
Potential Antimicrobial Activity of Galloyl-Flavonoid Glycosides From Woodfordia uniflora Against Methicillin-Resistant Staphylococcus aureus
Authors: Yu Jae Sik , Kim Ji-Hoon , Rashan Luay , Kim Inseo , Lee Wonsik , Kim Ki Hyun
Published: December 2021

Antibiotic-resistant infections are a growing problem; to combat multi-drug resistant bacterial infections, antibiotics with novel mechanisms of action are needed. Identification of potent bioactive natural products is an attractive avenue for developing novel therapeutic strategies against bacterial infections. As part of our ongoing research to explore bioactive natural products from diverse resources, we investigated the antimicrobial compounds fromWoodfordia uniflora, a flowering shrub unique to the Dhofar region of Oman. The plant has been used as a remedy for skin infections in Oman. However, to date, no study has examined the antimicrobial compounds inW. uniflora. Phytochemical analysis of the methanolic extract ofW. unifloraleaves in combination with LC/MS-based analysis allowed us to isolate and identify four flavonoid-type analogs (1–4), procyanidin B3-3-O-gallate (1), rhamnetin 3-O-(6″-galloyl)-β-D-glucopyranoside (2), rhamnetin 3-O-α-L-rhamnopyranoside (3), and quercetin 3-O-(6″-galloyl)-β-D-glucopyranoside (4). The isolates have a novel mechanism of action; the compounds inhibit biofilm formation in methicillin-resistantStaphylococcus aureus(MRSA) and synergize with methicillin. Our metabolite analysis revealed that this synergizing activity by compounds was achieved by remodeling metabolism including central carbon metabolism and glutamine biosynthesis that resulted in abnormal cell formation and reduction in biofilm formation of MRSA. Taken together, these findings provide experimental evidence that rhamnetin 3-O-(6″-galloyl)-β-D-glucopyranoside (2) and quercetin 3-O-(6″-galloyl)-β-D-glucopyranoside (4) can be considered as potential therapeutic agents for the treatment of methicillin-resistantS. aureus-associated diseases.
HSP27 Protein Dampens Encephalomyocarditis Virus Replication by Stabilizing Melanoma Differentiation-Associated Gene 5
Authors: Li Xiangrong , Ma Ruixian , Wu Bei , Niu Yuhui , Li Hongshan , Li Dianyu , Xie Jingying , Idris Adi , Feng Ruofei
Published: December 2021

Heat shock proteins (HSPs) are a protein family that respond to physiological stress, such as heat, starvation, and infection. As cellular protein chaperones, they play an important role in protein folding, assembly, and degradation. Though it is well known that HSP27 is involved in a range of viral infections, its role during an encephalomyocarditis virus (EMCV) infection is not known. Here, we report that EMCV degrades HSP27 and that EMCV proteins 2Cpro and 3Apro are primarily responsible for its degradation. Consequently, loss of cellular HSP27 augmented EMCV proliferation, an effect that could be reversed upon HSP27 overexpression. Importantly, we found that HSP27 positively regulated EMCV-triggered type I interferon (IFN) production. Moreover, overexpression of 2Cpro and 3Apro significantly blocked type I IFN production. We also found for the first time that HSP27, as a molecular chaperone, can specifically interact with MDA5 and stabilize the expression of MDA5. Collectively, this study shows that HSP27 dampens EMCV infectivity by positively regulating EMCV-triggered retinoic acid-inducible gene (RIG)-I-like receptor (RLR)/melanoma differentiation-associated gene 5 (MDA5) signal pathway, while EMCV proteins 2Cpro and 3Apro interact with HSP27 and degrade HSP27 protein expression to allow EMCV proliferation. Our findings provide further mechanistic evidence for EMCV partaking in immune escape mechanisms, and that 2Cpro and 3Apro could serve as potential antiviral targets.
Uses of Bacteriophages as Bacterial Control Tools and Environmental Safety Indicators
Authors: Rogovski Paula , Cadamuro Rafael Dorighello , da Silva Raphael , de Souza Estêvão Brasiliense , Bonatto Charline , Viancelli Aline , Michelon William , Elmahdy Elmahdy M. , Treichel Helen , Rodríguez-Lázaro David , Fongaro Gislaine
Published: December 2021

Bacteriophages are bacterial-specific viruses and the most abundant biological form on Earth. Each bacterial species possesses one or multiple bacteriophages and the specificity of infection makes them a promising alternative for bacterial control and environmental safety, as a biotechnological tool against pathogenic bacteria, including those resistant to antibiotics. This application can be either directly into foods and food-related environments as biocontrol agents of biofilm formation. In addition, bacteriophages are used for microbial source-tracking and as fecal indicators. The present review will focus on the uses of bacteriophages like bacterial control tools, environmental safety indicators as well as on their contribution to bacterial control in human, animal, and environmental health.