bluuebunny/crossref_metadata_2025 huggingface dataset:
Reduced Albumin Concentration Predicts Weight Gain and Higher Ad Libitum Energy Intake in Humans
Authors: Basolo Alessio , Ando Takafumi , Chang Douglas C. , Hollstein Tim , Krakoff Jonathan , Piaggi Paolo , Votruba Susanne
Published: March 2021

Objective

Circulating albumin is negatively associated with adiposity but whether it is associated with increased energy intake, lower energy expenditure or weight gain has not been examined.

Methods

In study 1 (n=238; 146 men), we evaluated whether fasting albumin concentration was associated with 24-h energy expenditure and ad libitum energy intake. In study 2 (n=325;167 men), we evaluated the association between plasma albumin and change in weight and body composition.

Results

After adjustment for known determinants of energy intake lower plasma albumin concentration was associated with greater total daily energy intake (β= 89.8 kcal/day per 0.1 g/dl difference in plasma albumin, p=0.0047). No associations were observed between plasma albumin concentrations and 24-h energy expenditure or 24-h respiratory quotient (p>0.2). Over 6 years, volunteers gained on average 7.5 ± 11.7 kg (p<0.0001). Lower albumin concentrations were associated with greater weight [β=3.53 kg, p=0.039 (adjusted for age, sex, follow up time), CI 0.16 to 6.21 per 1 g/dl difference albumin concentration] and fat mass (β=2.3 kg, p=0.022), respectively, but not with changes in fat free mass (p=0.06).

Conclusions

Lower albumin concentrations were associated with increased ad libitum food intake and weight gain, indicating albumin as a marker of energy intake regulation.

Clinical Trial Registration

ClinicalTrials.gov, identifiers NCT00340132, NCT00342732.
Effect of Tissue-Selective Estrogen Complex on Hip Structural Geometry in Postmenopausal Women: A 12-Month Study
Authors: Kim Bo Mi , Kim Sung Eun , Lee Dong-Yun , Choi DooSeok
Published: March 2021

Background

Hip structural analysis (HSA) is a method for evaluating bone geometry reflecting bone structural and biomechanical properties. However, tissue-selective estrogen complex (TSEC) treatment effects on HSA have not been investigated.

Objective

This study was performed to evaluate the effect of TSEC treatment on hip geometry in postmenopausal Korean women. The treatment was given for 12 months, and hip geometry was measured by HSA.

Materials and Methods

A total of 40 postmenopausal women who received TSEC containing conjugated estrogen 0.45 mg and bazedoxifene 20 mg for treating vasomotor symptoms were included in this retrospective cohort study. The changes in bone mineral density and parameters of HSA including the outer diameter, cross-sectional area, cross-sectional moment of inertia, cortical thickness, section modulus, and buckling ratio as determined by dual-energy X-ray absorptiometry were compared before and after 12 months of TSEC treatment.

Results

Mean age and years since menopause were 55.1 and 4.5 years, respectively. Total hip bone mineral density significantly increased by 0.74% after treatment (P=0.011). The changes in HSA were mainly demonstrated in the narrow femoral neck: cross-sectional area (P=0.003) and cortical thickness (P<0.001) increased significantly. For the shaft region, only SM decreased significantly after treatment (P=0.009). However, most parameters did not change significantly with TSEC treatment in the intertrochanteric and shaft regions.

Conclusions

Our findings demonstrate that 12 months of TSEC treatment could improve bone geometry as measured by HSA. The findings suggest that TSEC might be an interesting option for the prevention of fracture as well as osteoporosis in postmenopausal women.
RNA-Binding Proteins Play an Important Role in the Prognosis of Patients With Testicular Germ Cell Tumor
Authors: Yao Liangyu , Cong Rong , Ji Chengjian , Zhou Xiang , Luan Jiaochen , Meng Xianghu , Song Ninghong
Published: March 2021

Testicular germ cell tumors (TGCTs) are common urological neoplasms in young adult males. The outcome of TGCT depends on pathologic type and tumor stage. RNA-binding proteins (RBPs) influence numerous cancers via post-transcriptional regulation. The prognostic importance of RBPs in TGCT has not been fully investigated. In this study, we set up a prognostic risk model of TGCT using six significantly differentially expressed RBPs, namely, TRMT61A, POLR2J, DIS3L2, IFIH1, IGHMBP2, and NPM2. The expression profiles were downloaded from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression datasets. We observed by performing least absolute shrinkage and selection operator (LASSO) regression analyses that in the training cohort, the expression of six RBPs was correlated with disease-free survival in patients with TGCT. We assessed the specificity and sensitivity of 1-, 3-, 5-, and 10-year survival status prediction using receiver operating characteristic curve analysis and successfully validated using the test cohorts, the entire TCGA cohort, and Gene Expression Omnibus (GEO) datasets. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and gene set enrichment analyses were carried out to seek the possible signaling pathways related with risk score. We also examined the association between the model based on six RBPs and different clinical characteristics. A nomogram was established for TGCT recurrence prediction. Consensus clustering analysis was carried out to identify the clusters of TGCT with different clinical outcomes. Ultimately, external validations of the six-gene risk score were performed by using the GSE3218 and GSE10783 datasets downloaded from the GEO database. In general, our study constructed a prognostic model based on six RBPs, which could serve as independent risk factor in TGCT, especially in seminoma, and might have brilliant clinical application value.
A Novel Panel of 43 Insertion/Deletion Loci for Human Identifications of Forensic Degraded DNA Samples: Development and Validation
Authors: Jin Rui , Cui Wei , Fang Yating , Jin Xiaoye , Wang Hongdan , Lan Qiong , Guo Yuxin , Chen Chong , Zhang Xingru , Zhu Bofeng
Published: March 2021

Insertion/deletion polymorphism is a promising genetic marker in the forensic genetic fields, especially in the forensic application of degraded sample at crime scene. In this research, a novel five-dye multiplex amplification panel containing 43 highly polymorphic Insertion/deletion (InDel) loci and one Amelogenin gene locus is designed and constructed in-house for the individual identification in East Asian populations. The amplicon sizes of 43 InDel loci are less than 200 bp, which help to ensure that full allele profiles can be obtained from degraded DNA sample. A series of optimizations and developmental validations including optimization of PCR conditions, detection efficiency of the degraded and casework samples, sensitivity, reproducibility, precision, tolerance for inhibitors, species specificity and DNA mixtures are performed according to the Scientific Working Group on DNA Analysis Methods (SWGDAM) guideline. The results of the internal validation demonstrated that this novel InDel panel was a reliable, sensitive and accurate system with good tolerances to different inhibitors, and performed the considerable detection efficiency for the degraded or mixed samples, which could be used in the forensic applications.
Diet-Dependent Changes of the DNA Methylome Using a Göttingen Minipig Model for Obesity
Authors: Feng Y. , Cirera S. , Taşöz E. , Liu Y. , Olsen L. H. , Christoffersen B. Ø. , Pedersen H. D. , Ludvigsen T. P. , Kirk R. K. , Schumacher-Petersen C. , Deng Y. , Fredholm M. , Gao F.
Published: March 2021

Objective: Environmental factors can influence obesity by epigenetic mechanisms. The aim of this study was to investigate obesity-related epigenetic changes and the potential for reversal of these changes in the liver of Göttingen minipigs subjected to diet interventions.

Methods: High-throughput liquid hybridization capture-based bisulfite sequencing (LHC-BS) was used to quantify the methylation status of gene promotor regions in liver tissue in three groups of male castrated Göttingen minipigs: a standard chow group (SD, N = 7); a group fed high fat/fructose/cholesterol diet (FFC, N = 10) and a group fed high fat/fructose/cholesterol diet during 7 months and reversed to standard diet for 6 months (FFC/SD, N = 12). Expression profiling by qPCR of selected metabolically relevant genes was performed in liver tissue from all pigs.

Results: The pigs in the FFC diet group became morbidly obese. The FFC/SD diet did not result in a complete reversal of the body weight to the same weight as in the SD group, but it resulted in reversal of all lipid related metabolic parameters. Here we identified widespread differences in the patterning of cytosine methylation of promoters between the different feeding groups. By combining detection of differentially methylated genes with a rank-based hypergeometric overlap algorithm, we identified 160 genes showing differential methylation in corresponding promoter regions in the FFC diet group when comparing with both the SD and FFC/SD groups. As expected, this differential methylation under FFC diet intervention induced de-regulation of several metabolically-related genes involved in lipid/cholesterol metabolism, inflammatory response and fibrosis generation. Moreover, five genes, of which one is a fibrosis-related gene (MMP9), were still perturbed after diet reversion.

Conclusion: Our findings highlight the potential of exploring diet-epigenome interactions for treatment of obesity.
Immunological Hallmarks of Inflammatory Status in Vaso-Occlusive Crisis of Sickle Cell Anemia Patients
Authors: Silva-Junior Alexander Leonardo , Garcia Nadja Pinto , Cardoso Evilázio Cunha , Dias Stephanny , Tarragô Andrea Monteiro , Fraiji Nelson Abrahim , Gomes Matheus Souza , Amaral Laurence Rodrigues , Teixeira-Carvalho Andréa , Martins-Filho Olindo Assis , De Paula Erich Vinicius , Costa Allyson Guimarães , Malheiro Adriana
Published: March 2021

Sickle Cell Anemia (SCA) is the most common genetic disorder around the world. The mutation in the β-globin gene is responsible for a higher hemolysis rate, with further involvement of immunological molecules, especially cytokines, chemokines, growth factors, and anaphylatoxins. These molecules are responsible for inducing and attracting immune cells into circulation, thus contributing to increases in leukocytes and other pro-inflammatory mediators, and can culminate in a vaso-occlusive crisis (VOC). This study aimed to characterize the levels of these molecules in SCA patients in different clinical conditions in order to identify potential hallmarks of inflammation in these patients. An analytical prospective study was conducted using the serum of SCA patients in steady-state (StSt; n = 27) and VOC (n = 22), along with 53 healthy donors (HD). Samples from the VOC group were obtained on admission and on discharge, in the convalescent phase (CV). Levels of chemokines (CXCL8, CXCL10, CL2, CLL3, CCL4, CL5, and CCL11), cytokines (IL-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-10, IL-12p70, IL-13, IL-17A, TNF-α, and IFN-γ) and growth factors (VEGF, FGFb, PDGF-BB, GM-CSF, and G-CSF) were measured using a Luminex assay, and anaphylatoxins (C3a, C4a, and C5a) were measured using Cytometric Bead Array. SCA patients in StSt showed a pro-inflammatory profile, and were indicated as being higher producers of CCL2, IL-1β, IL-12p70, IFN-γ, IL-17A, and GM-CSF, while VOC is highlighted by molecules IL-4 and IL-5, but also IL-2, IL-7, PDGF-BB, and G-CSF. PDGF-BB and IL-1ra seemed to be two important hallmarks for the acute-to-chronic stage, due to their significant decrease after crisis inflammation and statistical difference in VOC and CV groups. These molecules show higher levels and a strong correlation with other molecules in VOC. Furthermore, they remain at higher levels even after crisis recovery, which suggest their importance in the role of inflammation during crisis and participation in immune cell adhesion and activation. These results support a relevant role of cytokines, neutrophil and monocytes, since these may act as markers of VOC inflammation in SCA patients.
Tenascin-C Deficiency Is Associated With Reduced Bacterial Outgrowth During Klebsiella pneumoniae-Evoked Pneumosepsis in Mice
Authors: Meijer Mariska T. , de Vos Alex F. , Scicluna Brendon P. , Roelofs Joris J. , Abou Fayçal Chérine , Orend Gertraud , Uhel Fabrice , van der Poll Tom
Published: March 2021

Tenascin C (TNC) is an extracellular matrix glycoprotein that recently emerged as an immunomodulator. TNC-deficient (TNC−/−) mice were reported to have a reduced inflammatory response upon systemic administration of lipopolysaccharide, the toxic component of gram-negative bacteria. Here, we investigated the role of TNC during gram-negative pneumonia derived sepsis. TNC+/+ and TNC−/− mice were infected with Klebsiella pneumoniae via the airways and sacrificed 24 and 42 h thereafter for further analysis. Pulmonary TNC protein levels were elevated 42 h after infection in TNC+/+ mice and remained undetectable in TNC−/− mice. TNC−/− mice showed modestly lower bacterial loads in lungs and blood, and a somewhat reduced local—but not systemic—inflammatory response. Moreover, TNC−/− and TNC+/+ mice did not differ with regard to neutrophil recruitment, lung pathology or plasma markers of distal organ injury. These results suggest that while TNC shapes the immune response during lipopolysaccharide-induced inflammation, this role may be superseded during pneumosepsis caused by a common gram-negative pathogen.
Identification and Phenotype of MAIT Cells in Cattle and Their Response to Bacterial Infections
Authors: Edmans Matthew D. , Connelley Timothy K. , Jayaraman Siddharth , Vrettou Christina , Vordermeier Martin , Mak Jeffrey Y. W. , Liu Ligong , Fairlie David P. , Maze Emmanuel Atangana , Chrun Tiphany , Klenerman Paul , Eckle Sidonia B. G. , Tchilian Elma , Benedictus Lindert
Published: March 2021

Mucosal-associated invariant T (MAIT) cells are a population of innate-like T cells that utilize a semi-invariant T cell receptor (TCR) α chain and are restricted by the highly conserved antigen presenting molecule MR1. MR1 presents microbial riboflavin biosynthesis derived metabolites produced by bacteria and fungi. Consistent with their ability to sense ligands derived from bacterial sources, MAIT cells have been associated with the immune response to a variety of bacterial infections, such asMycobacterium spp., Salmonella spp. and Escherichia coli. To date, MAIT cells have been studied in humans, non-human primates and mice. However, they have only been putatively identified in cattle by PCR based methods; no phenotypic or functional analyses have been performed. Here, we identified a MAIT cell population in cattle utilizing MR1 tetramers and high-throughput TCR sequencing. Phenotypic analysis of cattle MAIT cells revealed features highly analogous to those of MAIT cells in humans and mice, including expression of an orthologous TRAV1-TRAJ33 TCR α chain, an effector memory phenotype irrespective of tissue localization, and expression of the transcription factors PLZF and EOMES. We determined the frequency of MAIT cells in peripheral blood and multiple tissues, finding that cattle MAIT cells are enriched in mucosal tissues as well as in the mesenteric lymph node. Cattle MAIT cells were responsive to stimulation by 5-OP-RU and riboflavin biosynthesis competent bacteriain vitro. Furthermore, MAIT cells in milk increased in frequency in cows with mastitis. Following challenge with virulentMycobacterium bovis, a causative agent of bovine tuberculosis and a zoonosis, peripheral blood MAIT cells expressed higher levels of perforin. Thus, MAIT cells are implicated in the immune response to two major bacterial infections in cattle. These data suggest that MAIT cells are functionally highly conserved and that cattle are an excellent large animal model to study the role of MAIT cells in important zoonotic infections.
Previous History of American Tegumentary Leishmaniasis Alters Susceptibility and Immune Response Against Schistosoma mansoni Infection in Humans
Authors: Miranda Guilherme Silva , Resende Samira Diniz , Cardoso Diogo Tavares , Camelo Genil Mororó Araújo , Silva Jeferson Kelvin Alves Oliveira , Castro Vanessa Normandio de , Geiger Stefan Michael , Carneiro Mariângela , Negrão-Corrêa Deborah
Published: March 2021

Schistosomiasis and Leishmaniasis are chronic parasitic diseases with high prevalence in some tropical regions and, due to their wide distribution, a risk of co-infections is present in some areas. Nevertheless, the impact of this interaction on human populations is still poorly understood. Thus, the current study evaluated the effect of previous American Tegumentary Leishmaniasis (ATL) on the susceptibility and immune response toSchistosoma mansoniinfection in residents from a rural community in Northern of Minas Gerais state, Brazil, an area endemic for both parasitic infections. The participants answered a socioeconomic questionnaire and provided stool and blood samples for parasitological and immunological evaluations. Stool samples were examined by a combination of parasitological techniques to identify helminth infections, especiallyS. mansonieggs. Blood samples were used for hemograms and to measure the serum levels of cytokines and chemokines. Reports on previous ATL were obtained through interviews, clinical evaluation forms, and medical records.S. mansoniinfection was the most prevalent parasitic infection in the study population (46%), and the majority of the infected individuals had a very low parasite burden. In the same population, 93 individuals (36.2%) reported previous ATL, and the prevalence ofS. mansoniinfection among these individuals was significantly higher than among individuals with no ATL history. A multiple logistic regression model revealed thatS. mansoniinfection was positively associated with higher levels of CCL3 and CCL17, and a higher frequency of IL-17 responders. Moreover, this model demonstrated that individuals with an ATL history had a 2-fold higher probability to be infected withS. mansoni(OR = 2.0; 95% CI 1.04–3.68). AmongS. mansoni-infected individuals, the logistic regression demonstrated that a previous ATL history was negatively associated with the frequency of IL-17 responders and CXCL10 higher responders, but positively associated with higher IL-27 responders. Altogether, our data suggest that previous ATL may alter the susceptibility and the immune response inS. mansoni-infected individuals, which may likely affect the outcome of schistosomiasis and the severity of the disease in humans.
A Novel Sample Selection Approach to Aid the Identification of Factors That Correlate With the Control of HIV-1 Infection
Authors: Makinde Julia , Nduati Eunice W. , Freni-Sterrantino Anna , Streatfield Claire , Kibirige Catherine , Dalel Jama , Black S. Lucas , Hayes Peter , Macharia Gladys , Hare Jonathan , McGowan Edward , Abel Brian , King Deborah , Joseph Sarah ,   The IAVI Protocol C Investigators, Hunter Eric , Sanders Eduard J. , Price Matt , Gilmour Jill
Published: March 2021

Individuals infected with HIV display varying rates of viral control and disease progression, with a small percentage of individuals being able to spontaneously control infection in the absence of treatment. In attempting to define the correlates associated with natural protection against HIV, extreme heterogeneity in the datasets generated from systems methodologies can be further complicated by the inherent variability encountered at the population, individual, cellular and molecular levels. Furthermore, such studies have been limited by the paucity of well-characterised samples and linked epidemiological data, including duration of infection and clinical outcomes. To address this, we selected 10 volunteers who rapidly and persistently controlled HIV, and 10 volunteers each, from two control groups who failed to control (based on set point viral loads) from an acute and early HIV prospective cohort from East and Southern Africa. A propensity score matching approach was applied to control for the influence of five factors (age, risk group, virus subtype, gender, and country) known to influence disease progression on causal observations. Fifty-two plasma proteins were assessed at two timepoints in the 1st year of infection. We independently confirmed factors known to influence disease progression such as the B*57 HLA Class I allele, and infecting virus Subtype. We demonstrated associations between circulating levels of MIP-1α and IL-17C, and the ability to control infection. IL-17C has not been described previously within the context of HIV control, making it an interesting target for future studies to understand HIV infection and transmission. An in-depth systems analysis is now underway to fully characterise host, viral and immunological factors contributing to control.
Development of a Colloidal Gold-Based Immunochromatographic Strip for Rapid Detection of Severe Acute Respiratory Syndrome Coronavirus 2 Spike Protein
Authors: Li Ge , Wang Aiping , Chen Yumei , Sun Yaning , Du Yongkun , Wang Xun , Ding Peiyang , Jia Rui , Wang Yanwei , Zhang Gaiping
Published: March 2021

The outbreak and worldwide pandemic of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have a significant impact on global economy and human health. In order to reduce the disease spread, 16 monoclonal antibodies (McAbs) again SARS-CoV-2 were generated by immunized mice with the spike protein receptor binding domain (RBD), which was expressed in Chinese hamster ovary cell (CHO). A colloidal gold-based immunochromatographic strip was developed with two McAbs to detect SARS-CoV-2 spike protein, which can play a potential role in monitoring vaccine quality. The strip is highly specific, detecting only SARS-CoV-2 spike protein, and does not show any non-specific reactions with syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV) and other coronavirus and influenza viruses. The strip detected subunit vaccine in our laboratory with a detection limit of spike protein of 62.5 ng/mL. This strip provides an effective method in monitoring vaccine quality by detecting the antigen content of spike protein.
Immune Memory in Mild COVID-19 Patients and Unexposed Donors Reveals Persistent T Cell Responses After SARS-CoV-2 Infection
Authors: Ansari Asgar , Arya Rakesh , Sachan Shilpa , Jha Someshwar Nath , Kalia Anurag , Lall Anupam , Sette Alessandro , Grifoni Alba , Weiskopf Daniela , Coshic Poonam , Sharma Ashok , Gupta Nimesh
Published: March 2021

Understanding the causes of the diverse outcome of COVID-19 pandemic in different geographical locations is important for the worldwide vaccine implementation and pandemic control responses. We analyzed 42 unexposed healthy donors and 28 mild COVID-19 subjects up to 5 months from the recovery for SARS-CoV-2 specific immunological memory. Using HLA class II predicted peptide megapools, we identified SARS-CoV-2 cross-reactive CD4+ T cells in around 66% of the unexposed individuals. Moreover, we found detectable immune memory in mild COVID-19 patients several months after recovery in the crucial arms of protective adaptive immunity; CD4+ T cells and B cells, with a minimal contribution from CD8+ T cells. Interestingly, the persistent immune memory in COVID-19 patients is predominantly targeted towards the Spike glycoprotein of the SARS-CoV-2. This study provides the evidence of both high magnitude pre-existing and persistent immune memory in Indian population. By providing the knowledge on cellular immune responses to SARS-CoV-2, our work has implication for the development and implementation of vaccines against COVID-19.
Complement Genetic Variants and FH Desialylation in S. pneumoniae-Haemolytic Uraemic Syndrome
Authors: Gómez Delgado Irene , Corvillo Fernando , Nozal Pilar , Arjona Emilia , Madrid Álvaro , Melgosa Marta , Bravo Juan , Szilágyi Ágnes , Csuka Dorottya , Veszeli Nóra , Prohászka Zoltán , Sánchez-Corral Pilar
Published: March 2021

Haemolytic Uraemic Syndrome associated withStreptococcus pneumoniaeinfections (SP-HUS) is a clinically well-known entity that generally affects infants, and could have a worse prognosis than HUS associated toE. coliinfections. It has been assumed that complement genetic variants associated with primary atypical HUS cases (aHUS) do not contribute to SP-HUS, which is solely attributed to the action of the pneumococcal neuraminidase on the host cellular surfaces. We previously identified complement pathogenic variants and risk polymorphisms in a few Hungarian SP-HUS patients, and have now extended these studies to a cohort of 13 Spanish SP-HUS patients. Five patients presented rare complement variants of unknown significance, but the frequency of the risk haplotypes in theCFH-CFHR3-CFHR1region was similar to the observed in aHUS. Moreover, we observed desialylation of Factor H (FH) and the FH-Related proteins in plasma samples from 2 Spanish and 4 Hungarian SP-HUS patients. To analyze the functional relevance of this finding, we compared the ability of native and “in vitro” desialylated FH in: (a) binding to C3b-coated microtiter plates; (b) proteolysis of fluid-phase and surface-bound C3b by Factor I; (c) dissociation of surface bound-C3bBb convertase; (d) haemolytic assays on sheep erythrocytes. We found that desialylated FH had reduced capacity to control complement activation on sheep erythrocytes, suggesting a role for FH sialic acids on binding to cellular surfaces. We conclude that aHUS-risk variants in theCFH-CFHR3-CFHR1region could also contribute to disease-predisposition to SP-HUS, and that transient desialylation of complement FH by the pneumococcal neuraminidase may have a role in disease pathogenesis.
αB-Crystallin Alleviates Endotoxin-Induced Retinal Inflammation and Inhibits Microglial Activation and Autophagy
Authors: Wang Fangyu , Jiang Zhaoxin , Lou Bingsheng , Duan Fang , Qiu Suo , Cheng Zhixing , Ma Xinqi , Yang Yao , Lin Xiaofeng
Published: March 2021

αB-Crystallin, a member of the small heat shock protein (sHSP) family, plays an immunomodulatory and neuroprotective role by inhibiting microglial activation in several diseases. However, its effect on endotoxin-induced uveitis (EIU) is unclear. Autophagy may be associated with microglial activation, and αB-crystallin is involved in the regulation of autophagy in some cells. The role of αB-crystallin in microglial autophagy is unknown. This study aimed to explore the role of αB-crystallin on retinal microglial autophagy, microglial activation, and neuroinflammation in both cultured BV2 cells and the EIU mouse model. Our results show that αB-crystallin reduced the release of typical proinflammatory cytokines at both the mRNA and protein level, inhibited microglial activation in morphology, and suppressed the expression of autophagy-related molecules and the number of autophagolysosomes in vitro. In the EIU mouse model, αB-crystallin treatment alleviated the release of ocular inflammatory cytokines and the representative signs of inflammation, reduced the apoptosis of ganglion cells, and rescued retinal inflammatory structural and functional damage, as evaluated by optical coherence tomographic and electroretinography. Taken together, these results indicate that αB-crystallin inhibits the activation of microglia and supresses microglial autophagy, ultimately reducing endotoxin-induced neuroinflammation. In conclusion, αB-crystallin provides a novel and promising option for affecting microglial autophagy and alleviating symptoms of ocular inflammatory diseases.
Parameters of Capsulorrhexis and Intraocular Lens Decentration After Femtosecond and Manual Capsulotomies in High Myopic Patients With Cataracts
Authors: Zhu Yanan , Shi Kexin , Yao Ke , Wang Yuyan , Zheng Sifan , Xu Wen , Chen Peiqing , Yu Yibo , Shentu Xingchao
Published: March 2021

Purpose: To compare the parameters of capsulorrhexis and intraocular lens decentration after femtosecond laser capsulotomy and manual continuous curvilinear capsulorrhexis in high myopic patients with cataracts.

Methods: This is a prospective consecutive non-randomized comparative cohort study. Selected patients with axial length > 26.0 mm were divided into femtosecond laser capsulotomy (FS) group and manual continuous curvilinear capsulorrhexis (CCC) group. Five experienced phacoemulsification surgeons conducted all surgeries. Intraoperative complications and post-operative anterior segment photography were recorded. Intraocular lens decentration, area of capsulorrhexis, circularity, and capsule overlap were measured at 1 week, 1 month, and 2 years after surgery. Between group differences of parameters were determined with independent-sample t-test or the Mann–Whitney U-test, analysis of variance test, Pearson chi-square test, and Spearman rank correlation test.

Results: The study included 142 eyes (108 patients), 68 eyes in the FS group, and 74 eyes in the CCC group. At 1 week, 1 month, and 2 years after surgery, the area of capsulorrhexis in the CCC group was significantly larger than in the FS group (P < 0.05), while no significant difference was noted in circularity values. The complete overlap ratio in the FS group was significantly higher than that in the CCC group (P < 0.05) at each measured timepoint. Significant correlations were noted between the anterior chamber depth and the area of capsulorrhexis in the CCC group (R = 0.25, P = 0.04), but did not correlate in the FS group (P > 0.05). In patients with an anterior chamber depth >3 mm, the capsule-intraocular lens (IOL) overlap of the CCC group was less than that of the FS group at all measured timepoints after surgery (P < 0.05). Meanwhile, the IOL decentration in the CCC group was significantly greater than that of the FS group in those patients at 2 years after surgery (P < 0.05).

Conclusion: In high myopic patients with cataracts, with anterior chamber depth more than 3 mm, femtosecond laser capsulotomy can achieve better capsulorrhexis sizing and centering. Due to more precise capsulotomy and a better capsule-IOL overlap in the FS group, femtosecond laser capsulotomy resulted in better long-term centration of the IOL.
Age Matters: Community Assembly in the Pig Fecal Microbiome in the First Month of Life
Authors: Jurburg Stephanie D. , Bossers Alex
Published: March 2021

Despite the wealth of research into strategies for microbiome modulation, studies of microbiome management in pig hosts have found mixed results. A refined understanding of the patterns of microbiome assembly during the host’s early life, when management strategies are most commonly applied, is necessary for the development of successful management practices. Here, we study the development of the pig gut microbial community in a monitoring experiment, sampling the microbiome of pigs in a commercial farm intensively during the first month of life. We found that the community’s taxonomic richness increased linearly with host age. Furthermore, rapid changes across communities occurred in stages, and non-linear patterns in relative abundance were commonly observed among dominant taxa across host age, consistent with primary succession. Our results highlight the importance of understanding the patterns of microbiome assembly during host development, and identify successional stages as windows of opportunity for future research.
Multifaceted Interaction Between Hepatitis B Virus Infection and Lipid Metabolism in Hepatocytes: A Potential Target of Antiviral Therapy for Chronic Hepatitis B
Authors: Zhang Jiaxuan , Ling Ning , Lei Yu , Peng Mingli , Hu Peng , Chen Min
Published: March 2021

Hepatitis B virus (HBV) is considered a “metabolic virus” and affects many hepatic metabolic pathways. However, how HBV affects lipid metabolism in hepatocytes remains uncertain yet. Accumulating clinical studies suggested that compared to non-HBV-infected controls, chronic HBV infection was associated with lower levels of serum total cholesterol and triglycerides and a lower prevalence of hepatic steatosis. In patients with chronic HBV infection, high ALT level, high body mass index, male gender, or old age was found to be positively correlated with hepatic steatosis. Furthermore, mechanisms of how HBV infection affected hepatic lipid metabolism had also been explored in a number of studies based on cell lines and mouse models. These results demonstrated that HBV replication or expression induced extensive and diverse changes in hepatic lipid metabolism, by not only activating expression of some critical lipogenesis and cholesterolgenesis-related proteins but also upregulating fatty acid oxidation and bile acid synthesis. Moreover, increasing studies found some potential targets to inhibit HBV replication or expression by decreasing or enhancing certain lipid metabolism-related proteins or metabolites. Therefore, in this article, we comprehensively reviewed these publications and revealed the connections between clinical observations and experimental findings to better understand the interaction between hepatic lipid metabolism and HBV infection. However, the available data are far from conclusive, and there is still a long way to go before clarifying the complex interaction between HBV infection and hepatic lipid metabolism.
Effects of Essential Fatty Acid Supplementation on in vitro Fermentation Indices, Greenhouse Gas, Microbes, and Fatty Acid Profiles in the Rumen
Authors: Amanullah Sardar Muhammad , Kim Dong Hyeon , Paradhipta Dimas Hand Vidya , Lee Hyuk Jun , Joo Young Hoo , Lee Seong Shin , Kim Eun Tae , Kim Sam Churl
Published: March 2021

This study estimated the effect of essential fatty acid (FA) supplementation on fermentation indices, greenhouse gases, microbes, and FA profiles in the rumen. The treatments used pure FAs consisting of C18:2n-6 FA (LA), C18:3n-3 FA (LNA), or a mixture of these FAs at 1:1 ratio (Combo). In vitro rumen incubation was performed in 50 mL glass serum bottles containing 2 mg of pure FAs, 15 mL of rumen buffer (rumen fluid+anaerobe culture medium = 1:2), and 150 mg of synthetic diet (411 g cellulose, 411 g starch, and 178 g casein/kg dry matter) at 39°C for 8 h with five replications and three blanks. In rumen fermentation indices, LA exhibited highest (P < 0.05) ammonia-N and total gas volume after 8 h of incubation. Furthermore, LA presented lower (P < 0.05) pH with higher (P < 0.05) total volatile fatty acid (P = 0.034) than Combo, while LNA was not different compared with those in the other treatments. Additionally, Combo produced highest (P < 0.05) CO2 with lowest (P < 0.05) CH4. In the early hours of incubation, LA improved (P < 0.005) Fibrobacter succinogenes and Ruminococcus flavefaciens, while LNA improved (P < 0.005) Ruminococcus albus. After 8 h of incubation, LNA had lower (P < 0.05) methanogenic archaea than LA and Combo but had higher (P < 0.05) rumen ciliates than LA. R. albus was higher (P < 0.05) in LA than in LNA and Combo. It was observed that the rate of biohydrogenation of n-6 and n-3 FAs was comparatively lowest (P < 0.05) in Combo, characterized by higher C18:2n-6 and/or C18:3n-3 FA and polyunsaturated FA (PUFA) concentrations with lower (P < 0.05) concentrations of C18:0 and saturated FA and the ratio of saturated FAs to PUFAs. Therefore, this study concluded that dietary C18:2n-6 could improve populations of fibrolytic bacteria and rumen fermentation indices, but dietary mixture of pure C18:2n-6 and C18:3n-3 is recommended because it is effective in reducing enteric methane emissions and resisting biohydrogenation in the rumen with less effect on rumen microbes.
Identification and Characterization of EvpQ, a Novel T6SS Effector Encoded on a Mobile Genetic Element in Edwardsiella piscicida
Authors: Li Duan You , Liu Ying Li , Liao Xiao Jian , He Tian Tian , Sun Shan Shan , Nie Pin , Xie Hai Xia
Published: March 2021

In this study, a hypothetical protein (ORF02740) secreted by Edwardsiella piscicida was identified. We renamed the ORF02740 protein as EvpQ, which is encoded by a mobile genetic element (MGE) in E. piscicida genome. The evpQ gene is spaced by 513 genes from type VI secretion system (T6SS) gene cluster. Low GC content, three tRNA, and three transposase genes nearby evpQ define this MGE that evpQ localizes as a genomic island. Sequence analysis reveals that EvpQ shares a conserved domain of C70 family cysteine protease and shares 23.91% identity with T3SS effector AvrRpt2 of phytopathogenic Erwinia amylovora. Instead, EvpQ of E. piscicida is proved to be secreted at a T6SS-dependent manner, and it can be translocated into host cells. EvpQ is thereof a novel T6SS effector. Significantly decreased competitive index of ΔevpQ strain in blue gourami fish (0.53 ± 0.27 in head kidney and 0.44 ± 0.19 in spleen) indicates that EvpQ contributes to the pathogenesis of E. piscicida. At 8-, 18-, and 24-h post-subculture into DMEM, the transcription of evpQ was found to be negatively regulated by Fur and positively regulated by EsrC, and the steady-state protein levels of EvpQ are negatively controlled by RpoS. Our study lays a foundation for further understanding the pathogenic role of T6SS in edwardsiellosis.
Direct Observation of Conversion From Walled Cells to Wall-Deficient L-Form and Vice Versa in Escherichia coli Indicates the Essentiality of the Outer Membrane for Proliferation of L-Form Cells
Authors: Chikada Taiki , Kanai Tomomi , Hayashi Masafumi , Kasai Taishi , Oshima Taku , Shiomi Daisuke
Published: March 2021

Gram-negative bacteria such as Escherichia coli are surrounded by an outer membrane, which encloses a peptidoglycan layer. Even if thinner than in many Gram-positive bacteria, the peptidoglycan in E. coli allows cells to withstand turgor pressure in hypotonic medium. In hypertonic medium, E. coli treated with a cell wall synthesis inhibitor such as penicillin G form wall-deficient cells. These so-called L-form cells grow well under anaerobic conditions (i.e., in the absence of oxidative stress), becoming deformed and dividing as L-form. Upon removal of the inhibitor, they return to the walled rod-shaped state. Recently, the outer membrane was reported to provide rigidity to Gram-negative bacteria and to strengthen wall-deficient cells. However, it remains unclear why L-form cells need the outer membrane for growth. Using a microfluidic system, we found that, upon treatment with the outer membrane-disrupting drugs polymyxin B and polymyxin B nonapeptide or with the outer membrane synthesis inhibitor CHIR-090, the cells lysed during cell deformation and division, indicating that the outer membrane was important even in hypertonic medium. L-form cells could return to rod-shaped when trapped in a narrow space, but not in a wide space, likely due to insufficient physical force. Outer membrane rigidity could be compromised by lack of outer membrane proteins; Lpp, OmpA, or Pal. Deletion of lpp caused cells to lyse during cell deformation and cell division. In contrast, ompA and pal mutants could be deformed and return to small oval cells even when less physical force was exerted. These results strongly suggest that wall-deficient E. coli cells require a rigid outer membrane to survive, but not too rigid to prevent them from changing cell shape.
Prediction of Function Determining and Buried Residues Through Analysis of Saturation Mutagenesis Datasets
Authors: Bhasin Munmun , Varadarajan Raghavan
Published: March 2021

Mutational scanning can be used to probe effects of large numbers of point mutations on protein function. Positions affected by mutation are primarily at either buried or at exposed residues directly involved in function, hereafter designated as active-site residues. In the absence of prior structural information, it has not been easy to distinguish between these two categories of residues. We curated and analyzed a set of twelve published deep mutational scanning datasets. The analysis revealed differential patterns of mutational sensitivity and substitution preferences at buried and exposed positions. Prediction of buried-sites solely from the mutational sensitivity data was facilitated by incorporating predicted sequence-based accessibility values. For active-site residues we observed mean sensitivity, specificity and accuracy of 61, 90 and 88% respectively. For buried residues the corresponding figures were 59, 90 and 84% while for exposed non active-site residues these were 98, 44 and 82% respectively. We also identified positions which did not follow these general trends and might require further experimental re-validation. This analysis highlights the ability of deep mutational scans to provide important structural and functional insights, even in the absence of three-dimensional structures determined using conventional structure determination techniques, and also discuss some limitations of the methodology.
Cell Division Cycle Associated Genes as Diagnostic and Prognostic Biomarkers in Hepatocellular Carcinoma
Authors: Jiang Shan-Shan , Ke Sheng-Jie , Ke Zun-Li , Li Juan , Li Xiang , Xie Xing-Wei
Published: March 2021

With high mortality and poor prognosis, hepatocellular carcinoma (LIHC) has become the fourth leading cause of cancer-related deaths worldwide. Most of the LIHC patients missed the best treatment period because of the untimely diagnosis. For others, even if they are temporarily cured, they have to face a very low prognostic survival rate and a very high risk of recurrence. Based on the characteristics of abnormal proliferation and uncontrolled growth of tumor cells. Cell Division Cycle Associated (CDCA) family genes, which are responsible for regulating the cell cycle and proliferation, were selected as our research object to explore the mechanism of hepatocarcinogenesis. To this end, we investigated the expression profiles of CDCA family genes in LIHC and corresponding normal tissues, and the effect of CDCAs expression on the survival of prognosis and immune cell infiltration through bioinformatics analysis methods and the publicly accessible online databases. In addition, we also analyzed the expression correlation of CDCAs and screened the neighboring genes related to functional CDCAs. The results revealed that the expression levels of CDCA1/3/5/8 were significantly increased in LIHC, regardless of stage, sex, race, drinking behavior, and other clinical factors. CDCAs expression was significantly correlated with poor prognosis and was positively correlated with the infiltration of dendritic cells, B cells, and macrophages. We also found that the most relevant neighboring genes to CDCAs in LIHC were SGO2, NDC80, BIRC5, INCENP, and PLOD1. In general, our work suggests that CDCA1/3/5/8 has the potential to be a diagnostic gene in hepatocarcinogenesis and prognostic biomarkers for LIHC patients.
Sex-Specific Association of Endogenous PCSK9 With Memory Function in Elderly Subjects at High Cardiovascular Risk
Authors: Simeone Paola G. , Vadini Francesco , Tripaldi Romina , Liani Rossella , Ciotti Sonia , Di Castelnuovo Augusto , Cipollone Francesco , Santilli Francesca
Published: March 2021

Background: Growing evidence indicates that cognitive decline and cardiovascular diseases (CVDs) share common vascular risk factors. Protease proprotein convertase subtilisin/kexin type 9 (PCSK9) is associated with CV disease risk and has been also involved in neuronal differentiation.

Aim: Evaluate whether in patients at high CV risk cognitive function is related to PCSK9 levels.

Methods. One hundred sixty-six patients (67 female) were enrolled. A detailed neuropsychological (NP) assessment was performed. PCSK9 levels were measured with ELISA.

Results: Men had significantly higher short-term memory, executive function, and praxic and mental representation skills, as reflected by Forward Digit Span (FDS) (p = 0.005), Trail Making Test-A (TMT-A) (p = 0.047), Clock Drawing Test (CDT) (0.016). Endogenous PCSK9 levels were higher in female (p = 0.005). On linear regression analysis PCSK9 predicts short term memory only in females (Beta = 0.408, p = 0.001), with an interaction between PCSK9 and gender (p = 0.004 for interaction PCSK9 by sex). The association of PCSK9 with FDS in female was partially mediated by waist circumference (mediation effect 8.5%).

Conclusions: In patients at high CV risk short term memory was directly related to PCSK9 levels only in women, revealing the relevance of sex in this relationship. The association of PCSK9 with memory function may be mediated, at least in part, by waist circumference.
Circulating Inflamma-miRs as Potential Biomarkers of Cognitive Impairment in Patients Affected by Alzheimer’s Disease
Authors: Giuliani Angelica , Gaetani Simona , Sorgentoni Giulia , Agarbati Silvia , Laggetta Maristella , Matacchione Giulia , Gobbi Mirko , Rossi Tommaso , Galeazzi Roberta , Piccinini Gina , Pelliccioni Giuseppe , Bonfigli Anna Rita , Procopio Antonio Domenico , Albertini Maria Cristina , Sabbatinelli Jacopo , Olivieri Fabiola , Fazioli Francesca
Published: March 2021

Alzheimer’s disease (AD), the most prevalent neurodegenerative disease in the growing population of elderly people, is still lacking minimally-invasive circulating biomarkers that could facilitate the diagnosis and the monitoring of disease progression. MicroRNAs (miRNAs) are emerging as tissue-specific and/or circulating biomarkers of several age-related diseases, but evidence on AD is still not conclusive. Since a systemic pro-inflammatory status was associated with an increased risk of AD development and progression, we focused our investigation on a subset of miRNAs modulating the inflammatory process, namely inflamma-miRNAs. The expression of inflamma-miR-17-5p, -21-5p, -126-3p, and -146a-5p was analyzed in plasma samples from 116 patients with AD compared with 41 age-matched healthy control (HC) subjects. MiR-17-5p, miR-21-5p, and miR-126-3p plasma levels were significantly increased in AD patients compared to HC. Importantly, a strong inverse relationship was observed between miR-21-5p and miR-126-3p, and the cognitive impairment, assessed by Mini-Mental State Examination (MMSE). Notably, miR-126-3p was able to discriminate between mild and severe cognitive impairment. Overall, our results reinforce the hypothesis that circulating inflamma-miRNAs could be assessed as minimally invasive tools associated with the development and progression of cognitive impairment in AD.
Expression of Repressor Element 1 Silencing Transcription Factor (REST) in Serotonin Neurons in the Adult Male Nile Tilapia (Oreochromis niloticus)
Authors: Soga Tomoko , Nakajima Shingo , Parhar Ishwar S.
Published: March 2021

Repressor element-1 silencing transcription factor (REST) is highly expressed in the dorsal raphe where serotonin (5-hydroxytryptamine, 5-HT) neurons are located. REST works as a transcription factor for the 5-HT receptor and tryptophan hydroxylase two-gene expression. We hypothesized that REST is co-expressed in 5-HT neurons, which, if demonstrated, would be useful to understand the mechanism of 5-HT dysfunction-related disorders such as negative emotions and depression. Therefore, the present study was designed to examine the expression of the REST gene in the brain (forebrain, midbrain, and hindbrain) of adult male Nile tilapia (Oreochromis niloticus) using rt-PCR. Besides, using immunocytochemistry, co-localization of the REST gene was examined in 5-HT neurons and with neuronal-/glial-cell markers. We found a high expression of the REST gene in the midbrain region of the dorsal raphe, an area of 5-HT neurons. Double-label immunocytochemistry showed neuron-specific expression of REST co-localized in 5-HT neurons in the dorsal and ventral parts of the periventricular pretectal nucleus, paraventricular organ, and dorsal and medial raphe nucleus. Since midbrain 5-HT neurons express REST, we speculate that REST may control 5-HT neuronal activity related to negative emotions, including depression.