bluuebunny/crossref_metadata_2025 huggingface dataset:
Repressed Central Carbon Metabolism and Its Effect on Related Metabolic Pathways in Cefoperazone/Sulbactam-Resistant Pseudomonas aeruginosa
Authors: Chen Yue-tao , Yang Ke-xin , Dai Zhen-yuan , Yi Huan , Peng Xuan-xian , Li Hui , Chen Zhuang-gui
Published: March 2022

Metabolic shift and antibiotic resistance have been reported in Pseudomonas aeruginosa. However, the global metabolic characteristics remain largely unknown. The present study characterizes the central carbon metabolism and its effect on other metabolic pathways in cefoperazone-sulbactam (SCF)-resistant P. aeruginosa (PA-RSCF). GC-MS-based metabolomics shows a repressed central carbon metabolism in PA-RSCF, which is confirmed by measuring expression of genes and activity of enzymes in the metabolism. Furthermore, expression of the genes that encode the enzymes for the first step of fatty acid biosynthesis, glutamate metabolism, and electron transport chain is reduced, confirmed by their enzymatic activity assay, and the key enzyme for riboflavin metabolism is also reduced, indicating the decreased metabolic flux to the four related metabolic pathways. Moreover, the role of the reduced riboflavin metabolism, being related to ROS generation, in SCF resistance is explored. Exogenous H2O2 potentiates SCF-mediated killing in a dose-dependent manner, suggesting that the decreased ROS resulted from the reduced riboflavin metabolism that contributed to the resistance. These results indicate that the repressed central carbon metabolism and related riboflavin metabolism contribute to SCF resistance, but increasing ROS can restore SCF sensitivity. These findings characterize the repressed central carbon metabolism and its effect on other metabolic pathways as the global metabolic features in PA-RSCF.
Enabling Efficient Genetic Manipulations in a Rare Actinomycete Pseudonocardia alni Shahu
Authors: Li Jie , Wang Baiyang , Yang Qing , Si Han , Zhao Yuting , Zheng Yanli , Peng Wenfang
Published: March 2022

Pseudonocardia species are emerging as important microorganisms of global concern with unique and increasingly significant ecological roles and represent a prominent source of bioactive natural products, but genetic engineering of these organisms for biotechnological applications is greatly hindered due to the limitation of efficient genetic manipulation tools. In this regard, we report here the establishment of an efficient genetic manipulation system for a newly isolated strain, Pseudonocardia alni Shahu, based on plasmid conjugal transfer from Escherichia coli to Pseudonocardia. Conjugants were yielded upon determining the optimal ratio between the donor and recipient cells, and designed genome modifications were efficiently accomplished, including exogenous gene integration based on an integrative plasmid and chromosomal stretch removal by homologous recombination using a suicidal non-replicating vector. Collectively, this work has made the P. alni Shahu accessible for genetic engineering, and provided an important reference for developing genetic manipulation methods in other rare actinomycetes.
The N6-Methylandenosine-Related Gene BIRC5 as a Prognostic Biomarker Correlated With Cell Migration and Immune Cell Infiltrates in Low Grade Glioma
Authors: Jiang Xiulin , Shi Yulin , Chen Xi , Xu Haitao , Huang Xiaobin , Li Lihua , Pu Jun
Published: March 2022

Gliomas account for 75% of all primary malignant brain tumors in adults and are associated with high mortality. Emerging evidence has demonstrated that baculoviral inhibitor of apoptosis repeat containing 5 (BIRC5) plays a critical role in cell apoptosis and the progression of diverse cancers. However, no studies have yet focused on the immunological function and mechanisms of upstream BIRC5 regulation in the progression of low-grade gliomas (LGG). Here, we evaluated BIRC5 expression and clinical characteristics in people with LGG using the Chinese Glioma Genome Atlas, The Cancer Genome Atlas, Gene Expression Omnibus, Rembrandt, and Gravendeel databases. We used Kaplan–Meier statistics and receiver operating characteristic (ROC) curves to analyze the prognostic value of BIRC5 in LGG. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment terms were also explored to identify functional roles of BIRC5. The Tumor Immune Estimation Resource (TIMER) and Tumor Immune System Interaction (TISIDB) databases were used to examine the correlation between BIRC5 expression and immune cell infiltration in LGG. The Genomics of Drug Sensitivity in Cancer (GDSC) and Cancer Therapeutics Response Portal (CTRP) databases were used to examine the potential drugs targeting BIRC5. We used transwell and wound healing assays to determine the biological functions of BIRC5 in glioma cell migration. Our results demonstrated that BIRC5 was highly expressed in LGG and the expression level correlated with tumor grade, prognosis, histological subtype, isocitrate dehydrogenase 1 (IDH1) mutation, 1p/19q chromosomal co-deletion, chemotherapy status, and O[6]-methylguanine-DNA methyltransferase (MGMT) promoter methylation status. GO and KEGG analysis showed that BIRC5 is primarily involved in cell proliferation and immune response-related signaling pathways. We also found that BIRC5 was significantly correlated with m6A modification and diverse drug sensitivity. TIMER and TISIDB database analysis showed that BIRC5 expression is associated with infiltration of diverse immune cells and immune modulation in LGG. BIRC5 knockdown inhibited LGG cell migration. Collectively, our results demonstrate that BIRC5 is correlated with cell migration and immune infiltration in LGG and may be a useful prognostic biomarker.
Molecular Insights Into Binding and Activation of the Human KCNQ2 Channel by Retigabine
Authors: Garofalo Barbara , Bonvin Alexandre M.J.J. , Bosin Andrea , Di Giorgio Francesco P. , Ombrato Rosella , Vargiu Attilio V.
Published: March 2022

Voltage-gated potassium channels of the Kv7.x family are involved in a plethora of biological processes across many tissues in animals, and their misfunctioning could lead to several pathologies ranging from diseases caused by neuronal hyperexcitability, such as epilepsy, or traumatic injuries and painful diabetic neuropathy to autoimmune disorders. Among the members of this family, the Kv7.2 channel can form hetero-tetramers together with Kv7.3, forming the so-called M-channels, which are primary regulators of intrinsic electrical properties of neurons and of their responsiveness to synaptic inputs. Here, prompted by the similarity between the M-current and that in Kv7.2 alone, we perform a computational-based characterization of this channel in its different conformational states and in complex with the modulator retigabine. After validation of the structural models of the channel by comparison with experimental data, we investigate the effect of retigabine binding on the two extreme states of Kv7.2 (resting-closed and activated-open). Our results suggest that binding, so far structurally characterized only in the intermediate activated-closed state, is possible also in the other two functional states. Moreover, we show that some effects of this binding, such as increased flexibility of voltage sensing domains and propensity of the pore for open conformations, are virtually independent on the conformational state of the protein. Overall, our results provide new structural and dynamic insights into the functioning and the modulation of Kv7.2 and related channels.
The Consistency of Gastropod Identified Neurons Distinguishes Intra-Individual Plasticity From Inter-Individual Variability in Neural Circuits
Authors: Tamvacakis Arianna N. , Lillvis Joshua L. , Sakurai Akira , Katz Paul S.
Published: March 2022

Gastropod mollusks are known for their large, individually identifiable neurons, which are amenable to long-term intracellular recordings that can be repeated from animal to animal. The constancy of individual neurons can help distinguish state-dependent or temporal variation within an individual from actual variability between individual animals. Investigations into the circuitry underlying rhythmic swimming movements of the gastropod species,Tritonia exsulansandPleurobranchaea californicahave uncovered intra- and inter-individual variability in synaptic connectivity and serotonergic neuromodulation.Tritoniahas a reliably evoked escape swim behavior that is produced by a central pattern generator (CPG) composed of a small number of identifiable neurons. There is apparent individual variability in some of the connections between neurons that is inconsequential for the production of the swim behavior under normal conditions, but determines whether that individual can swim following a neural lesion. Serotonergic neuromodulation of synaptic strength intrinsic to the CPG creates neural circuit plasticity within an individual and contributes to reorganization of the network during recovery from injury and during learning. InPleurobranchaea, variability over time in the modulatory actions of serotonin and in expression of serotonin receptor genes in an identified neuron directly reflects variation in swimming behavior. Tracking behavior and electrophysiology over hours to days was necessary to identify the functional consequences of these intra-individual, time-dependent variations. This work demonstrates the importance of unambiguous neuron identification, properly assessing the animal and network states, and tracking behavior and physiology over time to distinguish plasticity within the same animal at different times from variability across individual animals.
Failure Of Hearing Acquisition in Mice With Reduced Expression of Connexin 26 Correlates With the Abnormal Phasing of Apoptosis Relative to Autophagy and Defective ATP-Dependent Ca2+ Signaling in Kölliker’s Organ
Authors: Sun Lianhua , Gao Dekun , Chen Junmin , Hou Shule , Li Yue , Huang Yuyu , Mammano Fabio , Chen Jianyong , Yang Jun
Published: March 2022

Mutations in the GJB2 gene that encodes connexin 26 (Cx26) are the predominant cause of prelingual hereditary deafness, and the most frequently encountered variants cause complete loss of protein function. To investigate how Cx26 deficiency induces deafness, we examined the levels of apoptosis and autophagy in Gjb2loxP/loxP; ROSA26CreER mice injected with tamoxifen on the day of birth. After weaning, these mice exhibited severe hearing impairment and reduced Cx26 expression in the cochlear duct. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) positive cells were observed in apical, middle, and basal turns of Kölliker’s organ at postnatal (P) day 1 (P1), associated with increased expression levels of cleaved caspase 3, but decreased levels of autophagy-related proteins LC3-II, P62, and Beclin1. In Kölliker’s organ cells with decreased Cx26 expression, we also found significantly reduced levels of intracellular ATP and hampered Ca2+ responses evoked by extracellular ATP application. These results offer novel insight into the mechanisms that prevent hearing acquisition in mouse models of non-syndromic hearing impairment due to Cx26 loss of function.
Diagnostic Challenges in Late Onset Multiple Acyl-CoA Dehydrogenase Deficiency: Clinical, Morphological, and Genetic Aspects
Authors: Lupica Antonino , Oteri Rosaria , Volta Sara , Ghezzi Daniele , Drago Selene Francesca Anna , Rodolico Carmelo , Musumeci Olimpia , Toscano Antonio
Published: March 2022

Background

Multiple acyl-CoA dehydrogenase deficiency (MADD) is an autosomal recessive disorder of fatty acid oxidation due to deficiency of the mitochondrial electron transfer chain. The late-onset form is characterized by exercise intolerance, muscle weakness, and lipid storage in myofibers. Most MADD patients greatly benefit from riboflavin supplementation.

Patients and methods

A retrospective study was conducted on patients with a diagnosis of vacuolar myopathy with lipid storage followed in our neuromuscular unit in the last 20 years. We selected 10 unrelated patients with the diagnosis of MADD according to clinical, morphological, and biochemical aspects. Clinical features, blood tests including serum acylcarnitines, EMG, and ENG were revised. Muscle biopsy was performed in all, and one individual underwent also a sural nerve biopsy. Gene sequencing of ETFA, ETFB, and ETFDH was performed as a first-tier genetic analysis followed by next-generation sequencing of an hyperCKemia gene panel in patients with undefined genotypes.

Results

Clinical evaluation at onset in all our patients showed fatigue and muscle weakness; four patients showed difficulties in chewing, three patients complained of dysphagia, two patients had a dropped head, and a patient had an unexpected ataxia with numbness and dysesthesia. Laboratory blood tests revealed a variable increase in serum CK (266–6,500) and LDH levels (500–2,000). Plasma acylcarnitine profile evidenced increased levels of different chains intermediates. EMG was either normal or showed myogenic or neurogenic patterns. NCS demonstrated sensory neuropathy in two patients. Muscle biopsies showed a vacuolar myopathy with a variable increase in lipid content. Nerve biopsy evidenced an axonal degeneration with the loss of myelinated fibers. ETFDH genetic analysis identifies 14 pathogenic variants. Patients were treated with high doses of riboflavin (400 mg/die). All of them showed a rapid muscle strength improvement and normalization of abnormal values in laboratory tests. Neuropathic symptoms did not improve.

Conclusion

Our data confirmed that clinical features in MADD patients are extremely variable in terms of disease onset and symptoms making diagnosis difficult. Laboratory investigations, such as serum acylcarnitine profile and muscle biopsy evaluation, may strongly address to a correct diagnosis. The favorable response to riboflavin supplementation strengthens the importance of an early diagnosis of these disorders among the spectrum of metabolic myopathies.
Verbal Training Induces Enhanced Functional Connectivity in Japanese Healthy Elderly Population
Authors: Yang Fan-Pei Gloria , Liu Tzu-Yu , Liu Chih-Hsuan , Murakami Shumei , Nakai Toshiharu
Published: March 2022

This study employs fMRI to examine the neural substrates of response to cognitive training in healthy old adults. Twenty Japanese healthy elders participated in a 4-week program and practiced a verbal articulation task on a daily basis. Functional connectivity analysis revealed that in comparison to age- and education-matched controls, elders who received the cognitive training demonstrated increased connectivity in the frontotemporal regions related with language and memory functions and showed significant correlations between the behavioral change in a linguistic task and connectivity in regions for goal-oriented persistence and lexical processing. The increased hippocampal connectivity was consistent with previous research showing efficacious memory improvement and change in hippocampal functioning. Moreover, the increased intra-network connectivity following cognitive training suggested an improved neural differentiation, in contrast to the inter-network activation pattern typical in the aging brain. This research not only validates the relationship of functional change in the frontal and temporal lobes to age-associated cognitive decline but also shows promise in turning neural change toward the right direction by cognitive training.
ERP Evidences of Rapid Semantic Learning in Foreign Language Word Comprehension
Authors: Soman Akshara , Ramachandran Prathibha , Ganapathy Sriram
Published: March 2022

The event-related potential (ERP) of electroencephalography (EEG) signals has been well studied in the case of native language speech comprehension using semantically matched and mis-matched end-words. The presence of semantic incongruity in the audio stimulus elicits a N400 component in the ERP waveform. However, it is unclear whether the semantic dissimilarity effects in ERP also appear for foreign language words that were learned in a rapid language learning task. In this study, we introduced the semantics of Japanese words to subjects who had no prior exposure to Japanese language. Following this language learning task, we performed ERP analysis using English sentences of semantically matched and mis-matched nature where the end-words were replaced with their Japanese counterparts. The ERP analysis revealed that, even with a short learning cycle, the semantically matched and mis-matched end-words elicited different EEG patterns (similar to the native language case). However, the patterns seen for the newly learnt word stimuli showed the presence of P600 component (delayed and opposite in polarity to those seen in the known language). A topographical analysis revealed that P600 responses were pre-dominantly observed in the parietal region and in the left hemisphere. The absence of N400 component in this rapid learning task can be considered as evidence for its association with long-term memory processing. Further, the ERP waveform for the Japanese end-words, prior to semantic learning, showed a P3a component owing to the subject's reaction to a novel stimulus. These differences were more pronounced in the centro-parietal scalp electrodes.
Synchronized Intracranial Electrical Activity and Gait Recording in Parkinson’s Disease Patients With Freezing of Gait
Authors: Liu De-Feng , Zhao Bao-Tian , Zhu Guan-Yu , Liu Yu-Ye , Bai Yu-Tong , Liu Huan-Guang , Jiang Yin , Zhang Xin , Lin-Shi  , Zhang Hua , Yang An-Chao , Zhang Jian-Guo
Published: March 2022

Background

This study aimed to describe a synchronized intracranial electroencephalogram (EEG) recording and motion capture system, which was designed to explore the neural dynamics during walking of Parkinson’s disease (PD) patients with freezing of gait (FOG). Preliminary analysis was performed to test the reliability of this system.

Methods

A total of 8 patients were enrolled in the study. All patients underwent bilateral STN-DBS surgery and were implanted with a right subdural electrode covering premotor and motor area. Synchronized electrophysiological and gait data were collected using the Nihon Kohden EEG amplifier and Codamotion system when subjects performed the Timed Up and Go (TUG) test. To verify the reliability of the acquisition system and data quality, we calculated and compared the FOG index between freezing and non-freezing periods during walking. For electrophysiological data, we first manually reviewed the scaled (five levels) quality during waking. Spectra comprising broadband electrocorticography (ECoG) and local field potential (LFP) were also compared between the FOG and non-FOG states. Lastly, connectivity analysis using coherence between cortical and STN electrodes were conducted. In addition, we also use machine learning approaches to classified FOG and non-FOG.

Results

A total of 8 patients completed 41 walking tests, 30 of which had frozen episodes, and 21 of the 30 raw data were level 1 or 2 in quality (70%). The mean ± SD walking time for the TUG test was 85.94 ± 47.68 s (range: 38 to 190.14 s); the mean ± SD freezing duration was 12.25 ± 7.35 s (range: 1.71 to 27.50 s). The FOG index significantly increased during the manually labeled FOG period (P< 0.05). The beta power of STN LFP in the FOG period was significantly higher than that in the non-FOG period (P< 0.05), while the band power of ECoG did not exhibit a significant difference between walking states. The coherence between the ECoG and STN LFP was significantly greater in high beta and gamma bands during the FOG period compared with the shuffled surrogates (P< 0.05). Lastly, STN-LFP band power features showed above-chance performance (p< 0.01, permutation test) in identifying FOG epochs.

Conclusion

In this study, we established and verified the synchronized ECoG/LFP and gait recording system in PD patients with FOG. Further neural substrates underlying FOG could be explored using the current system.
Improving Imaging of the Brainstem and Cerebellum in Autistic Children: Transformation-Based High-Resolution Diffusion MRI (TiDi-Fused) in the Human Brainstem
Authors: Guerrero-Gonzalez Jose , Surgent Olivia , Adluru Nagesh , Kirk Gregory R. , Dean III Douglas C. , Kecskemeti Steven R. , Alexander Andrew L. , Travers Brittany G.
Published: March 2022

Diffusion-weighted magnetic resonance imaging (dMRI) of the brainstem is technically challenging, especially in young autistic children as nearby tissue-air interfaces and motion (voluntary and physiological) can lead to artifacts. This limits the availability of high-resolution images, which are desirable for improving the ability to study brainstem structures. Furthermore, inherently low signal-to-noise ratios, geometric distortions, and sensitivity to motion not related to molecular diffusion have resulted in limited techniques for high-resolution data acquisition compared to other modalities such as T1-weighted imaging. Here, we implement a method for achieving increased apparent spatial resolution in pediatric dMRI that hinges on accurate geometric distortion correction and on high fidelity within subject image registration between dMRI and magnetization prepared rapid acquisition gradient echo (MPnRAGE) images. We call this post-processing pipeline T1 weighted-diffusion fused, or “TiDi-Fused”. Data used in this work consists of dMRI data (2.4 mm resolution, corrected using FSL’s Topup) and T1-weighted (T1w) MPnRAGE anatomical data (1 mm resolution) acquired from 128 autistic and non-autistic children (ages 6–10 years old). Accurate correction of geometric distortion permitted for a further increase in apparent resolution of the dMRI scan via boundary-based registration to the MPnRAGE T1w. Estimation of fiber orientation distributions and further analyses were carried out in the T1w space. Data processed with the TiDi-Fused method were qualitatively and quantitatively compared to data processed with conventional dMRI processing methods. Results show the advantages of the TiDi-Fused pipeline including sharper brainstem gray-white matter tissue contrast, improved inter-subject spatial alignment for group analyses of dMRI based measures, accurate spatial alignment with histology-based imaging of the brainstem, reduced variability in brainstem-cerebellar white matter tracts, and more robust biologically plausible relationships between age and brainstem-cerebellar white matter tracts. Overall, this work identifies a promising pipeline for achieving high-resolution imaging of brainstem structures in pediatric and clinical populations who may not be able to endure long scan times. This pipeline may serve as a gateway for feasibly elucidating brainstem contributions to autism and other conditions.
Changes in Food Purchasing Practices of French Households During the First COVID-19 Lockdown and Associated Individual and Environmental Factors
Authors: Recchia Daisy , Rollet Pascaline , Perignon Marlène , Bricas Nicolas , Vonthron Simon , Perrin Coline , Méjean Caroline
Published: March 2022

Background

To limit the spread of COVID-19, a strict lockdown was imposed in France between March and May 2020. Mobility limitations and closure of non-essential public places (restaurants, open-air markets, etc.) affected peoples' food environment (FE) and thus their food purchasing practices (FPPs). This study aimed to explore changes in FPPs of French households during lockdown and associations with individual and environmental factors.

Methods

In April of 2020 households from the Mont'Panier cross-sectional study (n = 306), a quota sampling survey conducted in the south of France, were asked to complete an online questionnaire about their FPPs during lockdown and related factors, including perceived FE (distance to closest general food store, perception of increased food prices, etc.). Objective FE (presence, number, proximity, and density of food outlets) was assessed around participant's home using a geographical information system. Multiple correspondence analysis based on changes in frequency of use and quantity of food purchased by food outlet, followed by a hierarchical cluster analysis, resulted in the identification of clusters. Logistic regression models were performed to assess associations between identified clusters and household's sociodemographic characteristics, perceived, and objective FE.

Results

Five clusters were identified. Cluster “Supermarket” (38% of the total sample), in which households reduced frequency of trips, but increased quantity bought in supermarkets during lockdown, was associated with lower incomes and the perception of increased food prices. Cluster “E-supermarket” (12%), in which households increased online food shopping with pickup at supermarket, was associated with higher incomes. Cluster “Diversified” (22%), made up of households who reduced frequency of trips to diverse food outlet types, was associated with the perception of increased food prices. Cluster “Organic Food Store” (20%), in which households did not change frequency of trips, nor quantity purchased in organic food stores, was associated with being older (35–50 y vs. <35 y). Finally, cluster “Producer” (8%), which includes households who regularly purchased food from producers, but mostly reduced these purchases during lockdown, was associated with the presence of an organic food store within a 1-km walking distance around home.

Conclusion

This study highlighted diverse changes in FPPs during lockdown and overall more significant associations with perceived than with objective FE indicators.
Proteomic Analysis of the Protective Effect of Eriodictyol on Benzo(a)pyrene-Induced Caco-2 Cytotoxicity
Authors: Wang Chong , Zhao Fan , Bai Yun , Li Chunbao , Xu Xinglian , Kristiansen Karsten , Zhou Guanghong
Published: March 2022

We evaluated the possible protective effects of six polyphenols on benzo(a)pyrene (BaP)-induced cytotoxicity in Caco-2 cells. We show that treatment with quinic acid, ferulic acid, homovanillic acid, trolox and BaP decreased cell viability, whereas naringenin and eriodictyol affected viability in a bi-phasic manner with low concentrations decreasing viability whereas higher concentrations increase viability. Co-treatment with 20 μM eriodictyol or naringenin reduced BaP-induced cytotoxicity, including cell apoptosis, cell cycle progression, and oxidative stress. Our results show that the protective effect of eriodictyol was superior to that of naringenin. The potential protective mechanisms of eriodictyol on BaP-induced toxicity were investigated by proteomics. We identified 80 differentially expressed proteins (DEPs) with proteins associated with genetic information processing pathway representing the highest proportion and number of proteins responding to eriodictyol treatment, including key proteins such as RPA2, SNRPA, RAD23B, NUP155 and AARS. Our results provide new knowledge on how polyphenols may prevent BaP-induced carcinogenesis.
Targeting Induced Local Lesions in the Wheat DEMETER and DRE2 Genes, Responsible for Transcriptional Derepression of Wheat Gluten Proteins in the Developing Endosperm
Authors: Wen Nuan , Osorio Claudia E. , Brew-Appiah Rhoda A. T. , Mejías Jaime H. , Alam Tariq , Kashyap Samneet , Reinbothe Steffen , Reinbothe Christiane , Moehs Charles P. , von Wettstein Diter , Rustgi Sachin
Published: March 2022

Wheat is a major source of energy and nutrition worldwide, but it is also a primary cause of frequent diet-induced health issues, specifically celiac disease, for which the only effective therapy so far is strict dietary abstinence from gluten-containing grains. Wheat gluten proteins are grouped into two major categories: high-molecular-weight glutenin subunits (HMWgs), vital for mixing and baking properties, and gliadins plus low-molecular-weight glutenin subunits (LMWgs) that contain the overwhelming majority of celiac-causing epitopes. We put forth a hypothesis that eliminating gliadins and LMWgs while retaining HMWgs might allow the development of reduced-immunogenicity wheat genotypes relevant to most gluten-sensitive individuals. This hypothesis stems from the knowledge that the molecular structures and regulatory mechanisms of the genes encoding the two groups of gluten proteins are quite different, and blocking one group's transcription, without affecting the other's, is possible. The genes for gliadins and LMWgs have to be de-methylated by 5-methylcytosine DNA glycosylase/lyase (DEMETER) and an iron-sulfur (Fe-S) cluster biogenesis enzyme (DRE2) early during endosperm development to permit their transcription. In this study, a TILLING (Targeting Induced Local Lesions IN Genomes) approach was undertaken to identify mutations in the homoeologous DEMETER (DME) and DRE2 genes in common and durum wheat. Lines with mutations in these genes were obtained that displayed reduced content of immunogenic gluten proteins while retaining essential baking properties. Although our data at first glance suggest new possibilities for treating celiac disease and are therefore of medical and agronomical interest, it also shows that inducing mutations in the DME and DRE2 genes analyzed here affected pollen viability and germination. Hence there is a need to develop other approaches in the future to overcome this undesired effect.
Three Different Types of β-Glucans Enhance Cognition: The Role of the Gut-Brain Axis
Authors: Hu Minmin , Zhang Peng , Wang Ruiqi , Zhou Menglu , Pang Ning , Cui Xiaoying , Ge Xing , Liu Xiaomei , Huang Xu-Feng , Yu Yinghua
Published: March 2022

Background

Dietary fiber is fermented in the lower gastrointestinal tract, potentially impacting the microbial ecosystem and thus may improve elements of cognition and brain function via the gut-brain axis. β-glucans, soluble dietary fiber, have different macrostructures and may exhibit different effects on the gut-brain axis. This study aimed to compare the effects of β-glucans from mushroom, curdlan and oats bran, representing β-(1,3)/(1,6)-glucan, β-(1,3)-glucan or β-(1,3)/(1,4)-glucan, on cognition and the gut-brain axis.

Methods

C57BL/6J mice were fed with either control diet or diets supplemented with β-glucans from mushroom, curdlan and oats bran for 15 weeks. The cognitive functions were evaluated by using the temporal order memory and Y-maze tests. The parameters of the gut-brain axis were examined, including the synaptic proteins and ultrastructure and microglia status in the hippocampus and prefrontal cortex (PFC), as well as colonic immune response and mucus thickness and gut microbiota profiles.

Results

All three supplementations with β-glucans enhanced the temporal order recognition memory. Brain-derived neurotrophic factor (BDNF) and the post-synaptic protein 95 (PSD95) increased in the PFC. Furthermore, mushroom β-glucan significantly increased the post-synaptic thickness of synaptic ultrastructure in the PFC whilst the other two β-glucans had no significant effect. Three β-glucan supplementations decreased the microglia number in the PFC and hippocampus, and affected complement C3 and cytokines expression differentially. In the colon, every β-glucan supplementation increased the number of CD206 positive cells and promoted the expression of IL-10 and reduced IL-6 and TNF-α expression. The correlation analysis highlights that degree of cognitive behavior improved by β-glucan supplementations was significantly associated with microglia status in the hippocampus and PFC and the number of colonic M2 macrophages. In addition, only β-glucan from oat bran altered gut microbiota and enhanced intestinal mucus.

Conclusions

We firstly demonstrated long-term supplementation of β-glucans enhanced recognition memory. Comparing the effects of β-glucans on the gut-brain axis, we found that β-glucans with different molecular structures exhibit differentia actions on synapses, inflammation in the brain and gut, and gut microbiota. This study may shed light on how to select appropriate β-glucans as supplementation for the prevention of cognitive deficit or improving immune function clinically.
The Cell Cycle-Associated Protein CDKN2A May Promotes Colorectal Cancer Cell Metastasis by Inducing Epithelial-Mesenchymal Transition
Authors: Shi Wei-Kun , Li Yun-Hao , Bai Xue-Shan , Lin Guo-Le
Published: March 2022

Colorectal cancer (CRC) is a common gastrointestinal malignancy, and recurrence and metastasis contribute considerably to its high mortality. It is well known that the epithelial-mesenchymal transition (EMT) accelerates the rate of cancer cell dissemination and migration, thus promoting cancer metastasis. Targeted therapy is a common modality for cancer treatment, and it can play a role in inhibiting cancer progression. In this study, bioinformatics was used to search for genes associated with the prognosis of CRC. First, differential analysis was performed on colon and rectal cancer samples to obtain 2,840 and 3,177 differentially expressed genes (DEGs), respectively. A Venn diagram was then used to identify 262 overlapping genes from the two groups of DEGs and EMT-related genes. The overlapping genes were subjected to batch survival analysis and batch expression analysis successively, and nine genes were obtained whose high expression in CRC led to a poor prognosis. The least absolute shrinkage and selection operator (LASSO) prognostic model was then constructed to obtain the risk score formula. A nomogram was constructed to seek prognostic independent factors to obtain CDKN2A. Finally, CCK-8 assay, flow cytometry and western blotting assays were performed to analyze the cellular biological function of CDKN2A. The results showed that knockdown of CDKN2A expression inhibited HT-29 cell proliferation, promoted apoptosis and cell cycle progression, and affected the EMT process in CRC.
CCDC134 as a Prognostic-Related Biomarker in Breast Cancer Correlating With Immune Infiltrates
Authors: Huang Zhijian , Yang Linhui , Chen Jian , Li Shixiong , Huang Jing , Chen Yijie , Liu Jingbo , Wang Hongyan , Yu Hui
Published: March 2022

Background

The expression of Coiled-Coil Domain Containing 134(CCDC134) is up-regulated in different pan-cancer species. However, its prognostic value and correlation with immune infiltration in breast cancer are unclear. Therefore, we evaluated the prognostic role of CCDC134 in breast cancer and its correlation with immune invasion.

Methods

We downloaded the transcription profile of CCDC134 between breast cancer and normal tissues from the Cancer Genome Atlas (TCGA). CCDC134 protein expression was assessed by the Clinical Proteomic Cancer Analysis Consortium (CPTAC) and the Human Protein Atlas. Gene set enrichment analysis (GSEA) was also used for pathway analysis. Receiver operating characteristic (ROC) curve was used to differentiate breast cancer from adjacent normal tissues. Kaplan-Meier method was used to evaluate the effect of CCDC134 on survival rate. The protein-protein interaction (PPI) network is built from STRING. Function expansion analysis is performed using the ClusterProfiler package. Through tumor Immune Estimation Resource (TIMER) and tumor Immune System Interaction database (TISIDB) to determine the relationship between CCDC134 expression level and immune infiltration. CTD database is used to predict drugs that inhibit CCDC134 and PubChem database is used to determine the molecular structure of identified drugs.

Results

The expression of CCDC134 in breast cancer tissues was significantly higher than that of CCDC134 mRNA expression in adjacent normal tissues. ROC curve analysis showed that the AUC value of CCDC134 was 0.663. Kaplan-meier survival analysis showed that patients with high CCDC134 had a lower prognosis (57.27 months vs 36.96 months, P = 2.0E-6). Correlation analysis showed that CCDC134 mRNA expression was associated with tumor purity immune invasion. In addition, CTD database analysis identified abrine, Benzo (A) Pyrene, bisphenol A, Soman, Sunitinib, Tetrachloroethylene, Valproic Acid as seven targeted therapy drugs that may be effective treatments for seven targeted therapeutics. It may be an effective treatment for inhibiting CCDC134.

Conclusion

In breast cancer, upregulated CCDC134 is significantly associated with lower survival and immune infiltrates invasion. Our study suggests that CCDC134 can serve as a biomarker of poor prognosis and a potential immunotherapy target in breast cancer. Seven drugs with significant potential to inhibit CCDC134 were identified.
Advanced Maternal Age Impairs Myelination in Offspring Rats
Authors: Han Wei , Pan Ya'nan , Han Ziyao , Cheng Li , Jiang Li
Published: March 2022

The effects of advanced maternal age (AMA) on the neurodevelopment of offspring are becoming increasingly important. Myelination is an important aspect of brain development; however, a limited number of studies have focused on the effects of AMA on myelination in offspring. The current study aims to evaluate the association between AMA and myelin sheath development in offspring. We studied the learning and memory function of immature offspring using the novel object recognition test. Then, we investigated the expression of myelin basic protein (MBP) in the immature offspring of young (3-month-old) and old (12-month-old) female rats at different time points (14, 28, and 60 days) after birth with immunofluorescence and western blotting. The myelin sheath ultrastructure was observed with transmission electron microscopy in immature and mature offspring. Extracellular signal-regulated kinase 1 and 2 (ERK1/2) and phosphorylated ERK1/2 (p-ERK1/2) were investigated by western blot in immature offspring at the above time points. AMA impaired the memory function of offspring during early postnatal days. The MBP expression level gradually increased with postnatal development in the offspring of both the AMA and Control (Ctl) groups, but the MBP level in the offspring of the AMA group was lower than that of the Ctl group at 14 days after birth. In addition, the ultrastructure of the myelin sheath was defective in AMA offspring during the early postnatal period; however, the myelin sheath was not significantly affected in offspring during adulthood. Interestingly, ERK phosphorylation at 14 days after birth was lower in AMA offspring than in Ctl offspring. However, ERK phosphorylation at 28 days after birth was higher in AMA offspring than in Ctl offspring. The peak of ERK phosphorylation in the AMA group was abnormal and delayed. Our results indicated that AMA is associated with poor developmental myelin formation in offspring. The ERK signaling pathway may play an essential role in the adverse effects of AMA on the offspring myelin sheath development.
A Retrospective Metabolomics Analysis of Gamma-Hydroxybutyrate in Humans: New Potential Markers and Changes in Metabolism Related to GHB Consumption
Authors: Wang Tingting , Nielsen Kirstine L. , Frisch Kim , Lassen Johan K. , Nielsen Camilla B. , Andersen Charlotte U. , Villesen Palle , Andreasen Mette F. , Hasselstrøm Jørgen B. , Johannsen Mogens
Published: March 2022

GHB is an endogenous short-chain organic acid presumably also widely applied as a rape and knock out drug in cases of drug-facilitated crimes or sexual assaults (DFSA). Due to the endogenous nature of GHB and its fast metabolism in vivo, the detection window of exogenous GHB is however narrow, making it challenging to prove use of GHB in DFSA cases. Alternative markers of GHB intake have recently appeared though none has hitherto been validated for forensic use. UHPLC-HRMS based screening of blood samples for drugs of abuse is routinely performed in several forensic laboratories which leaves an enormous amount of unexploited data. Recently we devised a novel metabolomics approach to use archived data from such routine screenings for elucidating both direct metabolites from exogenous compounds, but potentially also regulation of endogenous metabolism and metabolites. In this paper we used UHPLC-HRMS data acquired over a 6-year period from whole blood analysis of 51 drivers driving under the influence of GHB as well as a matched control group. The data were analyzed using a metabolomics approach applying a range of advanced analytical methods such as OPLS-DA, LASSO, random forest, and Pearson correlation to examine the data in depth and demonstrate the feasibility and potential power of the approach. This was done by initially detecting a range of potential biomarkers of GHB consumption, some that previously have been found in controlled GHB studies, as well as several new potential markers not hitherto known. Furthermore, we investigate the impact of GHB intake on human metabolism. In aggregate, we demonstrate the feasibility to extract meaningful information from archived data here exemplified using GHB cases. Hereby we hope to pave the way for more general use of the principle to elucidate human metabolites of e.g. new legal or illegal drugs as well as for applications in more global and large scale metabolomics studies in the future.
Beyond Circulating Tumor Cell Enumeration: Cell-Based Liquid Biopsy to Assess Protein Biomarkers and Cancer Genomics Using the RareCyte® Platform
Authors: Kaldjian Eric P. , Ramirez Arturo B. , Costandy Lillian , Ericson Nolan G. , Malkawi Walla I. , George Thaddeus C. , Kasi Pashtoon Murtaza
Published: March 2022

The practice of medicine has steadily employed less invasive methods to obtain information derived from the tumor to guide clinical management of patients. Liquid biopsy—the sampling of blood—is a non-invasive method for generating information previously only available from tissue biopsies of the tumor mass. Analysis of fragmented circulating tumor DNA in the plasma is clinically used to identify actionable mutations and detect residual or recurrent disease. Plasma analysis cannot, however, assess cancer phenotypes, including the expression of drug targets and protein biomarkers. Circulating tumor cells (CTCs) are intact cancer cells that have entered the blood that have the potential for distant metastasis. While enumeration of CTCs is prognostic of outcome, recently developed technology allows for the interrogation of protein biomarkers on CTCs that could be predictive of response. Furthermore, since CTCs contain intact whole cancer genomes, isolating viable CTCs detected during therapy could provide a rational approach to assessing mutational profiles of resistance. Identification, characterization and molecular analysis of CTCs together will advance the capacity of liquid biopsy to meet the requirements of twenty-first century medicine.
Tofacitinib Decreases Autophagy of Fibroblast-Like Synoviocytes From Rheumatoid Arthritis Patients
Authors: Vomero M. , Caliste M. , Barbati C. , Speziali M. , Celia A. I. , Ucci F. , Ciancarella C. , Putro E. , Colasanti T. , Buoncuore G. , Corsiero E. , Bombardieri M. , Spinelli F. R. , Ceccarelli F. , Conti F. , Alessandri C.
Published: March 2022

The pathway of Janus tyrosine kinases (JAKs) has a central role in the pathogenesis of Rheumatoid Arthritis (RA) by regulating multiple immune functions and cytokine production. The JAK inhibitor tofacitinib is effective in RA patients not responding to methotrexate or TNF-inhibitors. Since hyperactive autophagy has been associated with impaired apoptosis of RA fibroblast-like synoviocytes (FLS), we aimed to investigate the role of tofacitinib in modulating autophagy and apoptosis in these cells. FLS isolated from RA biopsies were cultured with tofacitinib in presence of autophagy inducer rapamycin and in serum deprivation condition. Levels of autophagy, apoptosis, and citrullinated proteins were analyzed by western blot, flow cytometry, immunocytofluorescence, and Real-Time PCR. Rapamycin induced an increase in RA-FLS autophagy while the levels of autophagy marker LC3-II were reduced after in vitro treatment with tofacitinib. The analysis of autophagic flux by specific fluorescence dye confirmed the reduction of autophagy in RA FLS. The treatment with tofacitinib did not influence apoptosis of RA FLS. Modulation of the autophagic process by tofacitinib did not significantly change citrullination. The results of this study demonstrate that tofacitinib is able to modulate autophagy of FLS contributing to its effectiveness in RA patients.
Experimental Investigation of Mechanical Characterizations of a POM Fiber-Reinforced Mortar Material
Authors: Zhang Jiuchang , Liu Tianxi , Dong Bo , Zhou Suhua , Guo Rongxin
Published: March 2022

In this study, a new fiber: polyoxymethylene fiber (POMF), was added into cement mortar to make a POM fiber-reinforced mortar (POMF-RM) material. When designing the mix proportion of POMF-RM, two factors were considered: fiber length (4, 8 and 12 mm) and dosage of POMF (volume fraction 0.24, 0.72 and 1.45%). When making the POMF-RM material, it was found that POMFs were easily dispersed into the mortar. Cubic and disc-shaped specimens of the POMF-RM material were made to conduct compression and Brazilian splitting experiments, respectively. It was found that, with increases in fiber length and volume content, the mechanical properties of the POMF-RM material were improved. Compared with the plain mortar, the elastic modulus, compressive and tensile strengths of the POMF-RM material significantly increased by 6.01, 13.18 and 5.34% at least, respectively. Among these two factors, the length of POMF played a more positive role. It was observed that the POMFs could effectively prevent the expansion of tensile cracks and significantly inhibit the complete disintegration of mortar fragments. This study showed that POMF was an excellent reinforcement material worthy of further research and application in practical engineering.
The Ratio of Oxygen Uptake From Ventilatory Anaerobic Threshold to Respiratory Compensation Point Is Maintained During Incremental Exercise in Older Adults
Authors: Kominami Kazuyuki , Imahashi Keiko , Katsuragawa Toko , Murakami Mitsuyo , Akino Masatoshi
Published: March 2022

Introduction

The period from ventilatory anaerobic threshold (VAT) to respiratory compensation point (RCP) during incremental exercise (isocapnic buffering phase) has been associated with exercise tolerance and skeletal muscle composition. However, several reports compare younger and older healthy adults, and specific age-related changes are unclear. This study aimed to examine the oxygen uptake (VO2) from VAT to RCP and its change over time in younger and older healthy adults.

Methods

A total of 126 consecutive participants were divided into two groups (95 younger and 31 older than 50 years of age) who underwent cardiopulmonary exercise testing, and VAT and RCP were determined. The ratio (RCP/VAT) and difference (ΔVO2 RCP-VAT) were calculated from the VO2 of VAT and RCP and compared between groups and ages. Statistical analyses included t-tests and Spearman’s correlation tests, and the significance level was set at <5%.

Results

RCP/VAT was not significantly different (1.40 ± 0.19 vs. 1.59 ± 0.24, p = 0.057) but weakly correlated with age (r = −0.229, p = 0.013, y = −0.0031x + 1.7588, lowering rate: 0.185%/year). Conversely, ΔVO2 RCP-VAT was significantly lower in the older group (7.7 ± 3.1 vs. 13.8 ± 4.9 ml/kg/min, p < 0.001) and correlated significantly with age (r = −0.499; p < 0.001; y = −0.1303x + 16.855; lowering rate, 0.914%/year).

Conclusion

ΔVO2 RCP-VAT was considered to be a poor indicator of lactate buffering capacity in the IB phase because both VAT and RCP were greatly affected by age-related decline. Conversely, RCP/VAT was suggested to be an index not easily affected by aging.