bluuebunny/crossref_metadata_2025 huggingface dataset:
Geographical Variation in Flower Color in the Grassland Daisy Gerbera aurantiaca: Testing for Associations With Pollinators and Abiotic Factors
Authors: Johnson Isabel M. , Edwards Trevor J. , Johnson Steven D.
Published: June 2021

Geographical variation in flower color of a plant species may reflect the outcome of selection by pollinators or may reflect abiotic factors such as soil chemistry or neutral processes such as genetic drift. Here we document striking geographical structure in the color of capitula of the endemic South African grassland daisyGerbera aurantiacaand ask which of these competing explanations best explains this pattern. The color of capitula ranges from predominantly red in the southwest to yellow in the center, with some northern populations showing within-population polymorphism. Hopliine scarab beetles were the most abundant flower visitors in all populations, apart from a yellow-flowered one where honeybees were frequent. In a mixed color population, yellow, orange and red morphs were equally attractive to hopliine beetles and did not differ significantly in terms of fruit set. Beetles were attracted to both red and yellow pan traps, but preferred the latter even at sites dominated by the red morph. We found no strong associations between morph color and abiotic factors, including soil chemistry. Plants in a common garden retained the capitulum color of the source population, even when grown from seed, suggesting that flower color variation is not a result of phenotypic plasticity. These results show that flower color inG. aurantiacais geographically structured, but the ultimate evolutionary basis of this color variation remains elusive.
Analysis of Age-Related Circular RNA Expression Profiles in Mesenchymal Stem Cells of Rat Bone Marrow
Authors: Sun Hui , Sun Yanan , Yu Xiao , Gao Xingyu , Wang Huan , Zhang Lin , Shi Yingai , He Xu
Published: June 2021

As multicellular organisms age, they undergo a reduction in tissue and organ function. Researchers have put forward a theory that stem cell aging is the main factor responsible for decreased tissue and organ function. The adult stem cells guarantee the maintenance and repair of adult tissues and organs. Among adult stem cells, mesenchymal stem cells (MSCs) are emerging as hopeful candidates for cell-based therapy of numerous diseases. In recent years, high-throughput sequencing technologies have evolved to identify circular RNAs (circRNAs) associated with an increasing number of diseases, such as cancer and age-related diseases. It has been reported that circRNAs can compete with microRNAs (miRNAs) to affect the stability or translation of target RNAs and further regulate gene expression at the transcriptional level. However, the role of circRNAs expressed in MSCs in aging mechanisms has not yet been deciphered. The aim of this study was to explore and analyze the expression profiles of age-related circRNAs in MSCs. In this study, bone marrow MSCs were extracted from aged and young rats and analyzed using high-throughput sequencing and bioinformatics. The reliability of high-throughput RNA sequencing was verified by quantitative real-time polymerase chain reaction. The most important circRNA functions and pathways were further selected by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomics (KEGG) analysis. Age-related circRNAs were found in the circrNA–miRNA–mRNA interaction network. The results of high-throughput sequencing showed that 4,229 circRNAs were involved in age-related senescence of MSCs. Compared with the young group, there were 29 differentially expressed circRNAs in the aged group, of which four were upregulated and 25 were downregulated. GO analysis covered three domains: biological process (BP), cellular component (CC), and molecular function (MF). The terms assigned to the BP domain were cellular metabolic processes and cellular macromolecule metabolic processes. The identified CC terms were intracellular and intracellular part, and the identified MF terms were binding and protein binding. The top five KEGG pathways were mitophagy–animal–Rattus norvegicus, prostate cancer–Rattus norvegicus, pathways in cancer–Rattus norvegicus, lysosome–Rattus norvegicus, and autophagy–animal–Rattus norvegicus. Altogether, circRNAs may play a major role in age-related MSC senescence. This study provides new mechanistic insights into MSC senescence, possibly leading to novel therapeutic strategies for age-related diseases.
A Platform for Validating Colorectal Cancer Driver Genes Using Mouse Organoids
Authors: Takeda Haruna
Published: June 2021

Systematic approaches for functionally validating cancer genes are needed since numerous genes mutated in cancer tissues have been identified from cancer genome sequencing. The mouse organoid culture system has been extensively used in the field of cancer research since mouse organoids can faithfully recapitulate the physiological behavior of the cells. Taking advantage of this, we recently described a platform for functionally validating colorectal cancer (CRC) driver genes that utilized CRISPR-Cas9 in mouse intestinal tumor organoids. In this review, we will describe how mouse organoids have been applied to CRC research and focus on how CRC genes can be validated using mouse organoids.
High-Resolution Quantitative Mapping of Macaque Cervicovaginal Epithelial Thickness: Implications for Mucosal Vaccine Delivery
Authors: Vincent Kathleen L. , Frost Patrice A. , Motamedi Massoud , Dick Edward J. , Wei Jingna , Yang Jinping , White Robert , Gauduin Marie-Claire
Published: June 2021

Vaginal mucosal surfaces naturally offer some protection against sexually transmitted infections (STIs) including Human Immunodeficiency Virus-1, however topical preventative medications or vaccine designed to boost local immune responses can further enhance this protection. We previously developed a novel mucosal vaccine strategy using viral vectors integrated into mouse dermal epithelium to induce virus-specific humoral and cellular immune responses at the site of exposure. Since vaccine integration occurs at the site of cell replication (basal layer 100-400 micrometers below the surface), temporal epithelial thinning during vaccine application, confirmed with high resolution imaging, is desirable. In this study, strategies for vaginal mucosal thinning were evaluated noninvasively using optical coherence tomography (OCT) to map reproductive tract epithelial thickness (ET) in macaques to optimize basal layer access in preparation for future effective intravaginal mucosal vaccination studies. Twelve adolescent female rhesus macaques (5-7kg) were randomly assigned to interventions to induce vaginal mucosal thinning, including cytobrush mechanical abrasion, the chemical surfactant spermicide nonoxynol-9 (N9), the hormonal contraceptive depomedroxyprogesterone acetate (DMPA), or no intervention. Macaques were evaluated at baseline and after interventions using colposcopy, vaginal biopsies, and OCT imaging, which allowed for real-time in vivo visualization and measurement of ET of the mid-vagina, fornices, and cervix. P value ≤0.05 was considered significant. Colposcopy findings included pink, rugated tissue with variable degrees of white-tipped, thickened epithelium. Baseline ET of the fornices was thinner than the cervix and vagina (p<0.05), and mensing macaques had thinner ET at all sites (p<0.001). ET was decreased 1 month after DMPA (p<0.05) in all sites, immediately after mechanical abrasion (p<0.05) in the fornix and cervix, and after two doses of 4% N9 (1.25ml) applied over 14 hrs in the fornix only (p<0.001). Histological assessment of biopsied samples confirmed OCT findings. In summary, OCT imaging allowed for real time assessment of macaque vaginal ET. While varying degrees of thinning were observed after the interventions, limitations with each were noted. ET decreased naturally during menses, which may provide an ideal opportunity for accessing the targeted vaginal mucosal basal layers to achieve the optimum epithelial thickness for intravaginal mucosal vaccination.
Differential Cytokine Responses in Hospitalized COVID-19 Patients Limit Efficacy of Remdesivir
Authors: Chan Yi-Hao , Young Barnaby E. , Fong Siew-Wai , Ding Ying , Goh Yun Shan , Chee Rhonda Sin-Ling , Tan Seow-Yen , Kalimuddin Shirin , Tambyah Paul A. , Leo Yee-Sin , Ng Lisa F. P. , Lye David Chien , Renia Laurent
Published: June 2021

A significant proportion of COVID-19 patients will progress to critical illness requiring invasive mechanical ventilation. This accentuates the need for a therapy that can reduce the severity of COVID-19. Clinical trials have shown the effectiveness of remdesivir in shortening recovery time and decreasing progression to respiratory failure and mechanical ventilation. However, some studies have highlighted its lack of efficacy in patients on high-flow oxygen and mechanical ventilation. This study uncovers some underlying immune response differences between responders and non-responders to remdesivir treatment. Immunological analyses revealed an upregulation of tissue repair factors BDNF, PDGF-BB and PIGF-1, as well as an increase in ratio of Th2-associated cytokine IL-4 to Th1-associated cytokine IFN-γ. Serological profiling of IgG subclasses corroborated this observation, with significantly higher magnitude of increase in Th2-associated IgG2 and IgG4 responses. These findings help to identify the mechanisms of immune regulation accompanying successful remdesivir treatment in severe COVID-19 patients.
Heavy Metal, Rare Earth Element and Pb Isotope Dynamics in Mussels During a Depuration Experiment in the Gulf of Aqaba, Northern Red Sea
Authors: Benaltabet Tal , Gutner-Hoch Eldad , Torfstein Adi
Published: June 2021

Mussels are considered highly efficient marine biomonitors, tracing anthropogenic and natural variations in heavy metals and various organic compounds. While heavy metals depuration processes in biomonitors are of growing interest, less knowledge is available regarding their Pb isotopes and rare earth elements (REEs) accumulation-release dynamics, and their response to short-term anthropogenic and terrigenous perturbations. Here, we report the results of a relocation experiment where a group of mussels (Brachidontes pharaonis) were extracted from a contaminated lagoon in the Gulf of Aqaba, northern Red Sea, and placed in water tanks that were flushed continuously with fresh, uncontaminated seawater. Specimens were removed periodically from the water table over a period of 13 weeks and trace and REEs and Pb isotopic compositions were determined separately for mussel’s shells and soft tissues. The results display a clear decrease over time in the concentrations of various heavy metals and REEs in the soft tissue, in concert with a similar shift in the Pb isotopic compositions toward seawater values. By contrast, the elemental and Pb isotopic composition of the shell presents little change over time. Coupling between the Pb isotopic composition of corresponding soft tissue and shell samples allows back-calculation of the timing and magnitude of abrupt pollution events and presents a novel approach for monitoring short-term pollution events. Nevertheless, given the coastal setting of the studied samples, it is important to consider the effects of terrigenous material on the results. Accordingly, Al-normalized element concentrations, Pb isotopes and calculated Ce anomalies, are used to identify two distinct terrigenous end members controlling the contaminated lagoon and the pristine site. The study demonstrates the potential of using mussels as robust biomonitors of natural and anthropogenic environmental perturbations through the combination between elemental concentrations and the isotopic composition of Pb.
The Aspergillus niger Major Allergen (Asp n 3) DNA-Specific Sequence Is a Reliable Marker to Identify Early Fungal Contamination and Postharvest Damage in Mangifera indica Fruit
Authors: Martínez Jorge , Nevado Ander , Suñén Ester , Gabriel Marta , Vélez-del-Burgo Ainara , Sánchez Patricia , Postigo Idoia
Published: June 2021

The aim of this work was to study the value of the main allergen Asp n 3 of Aspergillus niger as a molecular marker of allergenicity and pathogenicity with the potential to be used in the identification of A. niger as a contaminant and cause of spoilage of Mangifera indica. Real-time polymerase chain reaction (RT-PCR) was used for the amplification of Asp n 3 gene. Two pairs of primers were designed: one for the amplification of the entire sequence and another one for the amplification of the most conserved region of this peroxisomal protein. The presence of A. niger was demonstrated by the early detection of the allergenic protein Asp n 3 coding gene, which could be considered a species-specific marker. The use of primers designed based on the conserved region of the Asp n 3 encoding gene allowed us to identify the presence of the closely related fungal species Aspergillus fumigatus by detecting Asp n 3 homologous protein, which can be cross-reactive. The use of conserved segments of the Asp n 3 gene or its entire sequence allows us to detect phylogenetically closely related species within the Aspergilaceae family or to identify species-specific contaminating fungi.
A Complementary Mechanism of Bacterial mRNA Translation Inhibition by Tetracyclines
Authors: Barrenechea Victor , Vargas-Reyes Maryhory , Quiliano Miguel , Milón Pohl
Published: June 2021

Tetracycline has positively impacted human health as well as the farming and animal industries. Its extensive usage and versatility led to the spread of resistance mechanisms followed by the development of new variants of the antibiotic. Tetracyclines inhibit bacterial growth by impeding the binding of elongator tRNAs to the ribosome. However, a small number of reports indicated that Tetracyclines could also inhibit translation initiation, yet the molecular mechanism remained unknown. Here, we use biochemical and computational methods to study how Oxytetracycline (Otc), Demeclocycline (Dem), and Tigecycline (Tig) affect the translation initiation phase of protein synthesis. Our results show that all three Tetracyclines induce Initiation Factor IF3 to adopt a compact conformation on the 30S ribosomal subunit, similar to that induced by Initiation Factor IF1. This compaction was faster for Tig than Dem or Otc. Furthermore, all three tested tetracyclines affected IF1-bound 30S complexes. The dissociation rate constant of IF1 in early 30S complexes was 14-fold slower for Tig than Dem or Otc. Late 30S initiation complexes (30S pre-IC or IC) exhibited greater IF1 stabilization by Tig than for Dem and Otc. Tig and Otc delayed 50S joining to 30S initiation complexes (30S ICs). Remarkably, the presence of Tig considerably slowed the progression to translation elongation and retained IF1 in the resulting 70S initiation complex (70S IC). Molecular modeling of Tetracyclines bound to the 30S pre-IC and 30S IC indicated that the antibiotics binding site topography fluctuates along the initiation pathway. Mainly, 30S complexes show potential contacts between Dem or Tig with IF1, providing a structural rationale for the enhanced affinity of the antibiotics in the presence of the factor. Altogether, our data indicate that Tetracyclines inhibit translation initiation by allosterically perturbing the IF3 layout on the 30S, retaining IF1 during 70S IC formation, and slowing the transition toward translation elongation. Thus, this study describes a new complementary mechanism by which Tetracyclines may inhibit bacterial protein synthesis.
Quorum Sensing and Cyclic di-GMP Exert Control Over Motility of Vibrio fischeri KB2B1
Authors: Dial Courtney N. , Eichinger Steven J. , Foxall Randi , Corcoran Christopher J. , Tischler Alice H. , Bolz Robert M. , Whistler Cheryl A. , Visick Karen L.
Published: June 2021

Bacterial motility is critical for symbiotic colonization by Vibrio fischeri of its host, the squid Euprymna scolopes, facilitating movement from surface biofilms to spaces deep inside the symbiotic organ. While colonization has been studied traditionally using strain ES114, others, including KB2B1, can outcompete ES114 for colonization for a variety of reasons, including superior biofilm formation. We report here that KB2B1 also exhibits an unusual pattern of migration through a soft agar medium: whereas ES114 migrates rapidly and steadily, KB2B1 migrates slowly and then ceases migration. To better understand this phenomenon, we isolated and sequenced five motile KB2B1 suppressor mutants. One harbored a mutation in the gene for the cAMP receptor protein (crp); because this strain also exhibited a growth defect, it was not characterized further. Two other suppressors contained mutations in the quorum sensing pathway that controls bacterial bioluminescence in response to cell density, and two had mutations in the diguanylate cyclase (DGC) gene VF_1200. Subsequent analysis indicated that (1) the quorum sensing mutations shifted KB2B1 to a perceived low cell density state and (2) the high cell density state inhibited migration via the downstream regulator LitR. Similar to the initial point mutations, deletion of the VF_1200 DGC gene increased migration. Consistent with the possibility that production of the second messenger c-di-GMP inhibited the motility of KB2B1, reporter-based measurements of c-di-GMP revealed that KB2B1 produced higher levels of c-di-GMP than ES114, and overproduction of a c-di-GMP phosphodiesterase promoted migration of KB2B1. Finally, we assessed the role of viscosity in controlling the quorum sensing pathway using polyvinylpyrrolidone and found that viscosity increased light production of KB2B1 but not ES114. Together, our data indicate that while the two strains share regulators in common, they differ in the specifics of the regulatory control over downstream phenotypes such as motility.
Sexual Dimorphism in Aggression: Sex-Specific Fighting Strategies Across Species
Authors: Pandolfi Matias , Scaia Maria Florencia , Fernandez Maria Paz
Published: June 2021

Aggressive behavior is thought to have evolved as a strategy for gaining access to resources such as territory, food, and potential mates. Across species, secondary sexual characteristics such as competitive aggression and territoriality are considered male-specific behaviors. However, although female–female aggression is often a behavior that is displayed almost exclusively to protect the offspring, multiple examples of female–female competitive aggression have been reported in both invertebrate and vertebrate species. Moreover, cases of intersexual aggression have been observed in a variety of species. Genetically tractable model systems such as mice, zebrafish, and fruit flies have proven extremely valuable for studying the underlying neuronal circuitry and the genetic architecture of aggressive behavior under laboratory conditions. However, most studies lack ethological or ecological perspectives and the behavioral patterns available are limited. The goal of this review is to discuss each of these forms of aggression, male intrasexual aggression, intersexual aggression and female intrasexual aggression in the context of the most common genetic animal models and discuss examples of these behaviors in other species.
Inhibition of Perforin-Mediated Neurotoxicity Attenuates Neurological Deficits After Ischemic Stroke
Authors: Pan Yuhualei , Tian Dan , Wang Huan , Zhao Yushang , Zhang Chengjie , Wang Song , Xie Dan , Zhang Dong , Zhu Yanbing , Zhang Yongbo
Published: June 2021

Perforin-mediated cytotoxicity plays a crucial role in microbial defense, tumor surveillance, and primary autoimmune disorders. However, the contribution of the cytolytic protein perforin to ischemia-induced secondary tissue damage in the brain has not been fully investigated. Here, we examined the kinetics and subpopulations of perforin-positive cells and then evaluated the direct effects of perforin-mediated cytotoxicity on outcomes after ischemic stroke. Using flow cytometry, we showed that perforin+CD45+ immune cells could be detected at 12 h and that the percentage of these cells increased largely until on day 3 and then significantly declined on day 7. Surprisingly, the percentage of Perforin+CD45+ cells also unexpectedly increased from day 7 to day 14 after ischemic stroke in Perforin1-EGFP transgenic mice. Our results suggested that Perforin+CD45+ cells play vital roles in the ischemic brain at early and late stages and further suggested that Perforin+CD45+ cells are a heterogeneous population. Surprisingly, in addition to CD8+ T cells, NK cells, and NKT cells, central nervous system (CNS)-resident immune microglia, which are first triggered and activated within minutes after ischemic stroke in mice, also secreted perforin during ischemic brain injury. In our study, the percentage of perforin+ microglia increased from 12 h after ischemic stroke, increased largely until on day 3 after ischemic stroke, and then moderately declined from days 3 to 7. Intriguingly, the percentage of perforin+ microglia also dramatically increased from days 7 to 14 after ischemic stroke. Furthermore, compared with wild-type littermates, Perforin 1–/– mice exhibited significant increases in the cerebral infarct volume, neurological deficits, and neurogenesis and inhibition of neurotoxic astrogliosis. Interestingly, the number of CD45+CD3+ T cells was significantly decreased in Perforin 1–/– mice compared with their wild-type littermates, especially the number of γδ T cells. In addition, Perforin 1–/– mice had lower levels of IL-17 than their wild-type littermates. Our results identified a critical function of perforin-mediated neurotoxicity in the ischemic brain, suggesting that targeting perforin-mediated neurotoxicity in brain-resident microglia and invading perforin+CD45+ immune cells may be a potential strategy for the treatment of ischemic stroke.
Functional Neuroimaging During Asleep DBS Surgery: A Proof of Concept Study
Authors: Sammartino Francesco , Taylor Paul , Chen Gang , Reynolds Richard C. , Glen Daniel , Krishna Vibhor
Published: June 2021

Object: A real-time functional magnetic resonance imaging (fMRI) feedback during ventral intermediate nucleus (VIM) deep brain stimulation (DBS) under general anesthesia (or “asleep” DBS) does not exist. We hypothesized that it was feasible to acquire a reliable and responsive fMRI during asleep VIM DBS surgery.

Methods: We prospectively enrolled 10 consecutive patients who underwent asleep DBS for the treatment of medication-refractory essential tremor. Under general anesthesia, we acquired resting-state functional MRI immediately before and after the cannula insertion. Reliability was determined by a temporal signal-to-noise-ratio >100. Responsiveness was determined based on the fMRI signal change upon insertion of the cannula to the VIM.

Results: It was feasible to acquire reliable fMRI during asleep DBS surgery. The fMRI signal was responsive to the brain cannula insertion, revealing a reduction in the tremor network's functional connectivity, which did not reach statistical significance in the group analysis.

Conclusions: It is feasible to acquire a reliable and responsive fMRI signal during asleep DBS. The acquisition steps and the preprocessing pipeline developed in these experiments will be useful for future investigations to develop fMRI-based feedback for asleep DBS surgery.
White Matter Hyperintensities Are Associated With Severity of Essential Tremor in the Elderly
Authors: Becktepe Jos S. , Busse Johannes , Jensen-Kondering Ulf , Toedt Inken , Wolff Stephan , Zeuner Kirsten E. , Berg Daniela , Granert Oliver , Deuschl Günther
Published: June 2021

Background: Essential tremor (ET) occurs with steeply increasing prevalence in the elderly, and apart from disease duration, age is independently associated with an increase of tremor amplitude and a decrease of frequency. White matter hyperintensities (WMHs) are a common finding in the elderly, and their role in the pathophysiology of ET is unknown. The aims of this study were to examine whether ET patients differ in their total or region-specific WMH volumes from healthy controls and to determine the impact of WMH on tremor characteristics.

Methods: A total of 47 elderly ET patients with a mean age of 72 years and 39 age-matched healthy controls underwent a thorough clinical assessment and 3T MRI. Total WMH volumes were derived from T2-weighted fluid-attenuated inversion recovery (FLAIR) MR images. Additionally, region of interest-based WMH volumes for the Johns Hopkins University (JHU) white matter tracts and labels were calculated, and WMHs were assessed semiquantitatively using the Fazekas scale.

Results: Essential tremor patients and healthy controls did not differ in their total or tract-specific WMH volumes or Fazekas scores. However, WMH volume was significantly positively correlated with tremor severity on the TETRAS scale, and there was a significant negative correlation with the mean accelerometric tremor frequency. In a multiple linear regression model including disease duration, age, and age-adjusted total WMH volume, only the WMH volume significantly predicted tremor severity, while age and disease duration were not significant.

Conclusion: We found evidence for a direct association between WMH volume and tremor severity. If confirmed by larger studies, our findings could explain the well-known relation between age and tremor severity.
Pathophysiology of Cerebellar Tremor: The Forward Model-Related Tremor and the Inferior Olive Oscillation-Related Tremor
Authors: Kakei Shinji , Manto Mario , Tanaka Hirokazu , Mitoma Hiroshi
Published: June 2021

Lesions in the Guillain–Mollaret (G–M) triangle frequently cause various types of tremors or tremor-like movements. Nevertheless, we know relatively little about their generation mechanisms. The deep cerebellar nuclei (DCN), which is a primary node of the triangle, has two main output paths: the primary excitatory path to the thalamus, the red nucleus (RN), and other brain stem nuclei, and the secondary inhibitory path to the inferior olive (IO). The inhibitory path contributes to the dentato-olivo-cerebellar loop (the short loop), while the excitatory path contributes to the cerebrocerebellar loop (the long loop). We propose a novel hypothesis: each loop contributes to physiologically distinct type of tremors or tremor-like movements. One type of irregular tremor-like movement is caused by a lesion in the cerebrocerebellar loop, which includes the primary path. A lesion in this loop affects the cerebellar forward model and deteriorates its accuracy of prediction and compensation of the feedback delay, resulting in irregular instability of voluntary motor control, i.e., cerebellar ataxia (CA). Therefore, this type of tremor, such as kinetic tremor, is usually associated with other symptoms of CA such as dysmetria. We call this type of tremor forward model-related tremor. The second type of regular tremor appears to be correlated with synchronized oscillation of IO neurons due, at least in animal models, to reduced degrees of freedom in IO activities. The regular burst activity of IO neurons is precisely transmitted along the cerebellocerebral path to the motor cortex before inducing rhythmical reciprocal activities of agonists and antagonists, i.e., tremor. We call this type of tremor IO-oscillation-related tremor. Although this type of regular tremor does not necessarily accompany ataxia, the aberrant IO activities (i.e., aberrant CS activities) may induce secondary maladaptation of cerebellar forward models through aberrant patterns of long-term depression (LTD) and/or long-term potentiation (LTP) of the cerebellar circuitry. Although our hypothesis does not cover all tremors or tremor-like movement disorders, our approach integrates the latest theories of cerebellar physiology and provides explanations how various lesions in or around the G–M triangle results in tremors or tremor-like movements. We propose that tremor results from errors in predictions carried out by the cerebellar circuitry.
Geographical Factor Influences the Metabolite Distribution of House Edible Bird's Nests in Malaysia
Authors: Tong Shi-Ruo , Lee Ting-Hun , Cheong Soon-Keng , Lim Yang-Mooi
Published: June 2021

Background: Edible Bird's Nest (EBN) is famously consumed as a food tonic for its high nutritional values with numerous recuperative and therapeutic properties. EBN is majority exploited from swiftlet houses but the differences in terms of metabolite distribution between the production site of house EBN is not yet fully understood. Therefore, this study was designed to identify the metabolite distribution and to determine the relationship pattern for the metabolite distribution of house EBNs from different locations in Malaysia.

Methods: The differences of metabolite distribution in house EBN were studied by collecting the samples from 13 states in Malaysia. An extraction method of eHMG was acquired to extract the metabolites of EBN and was subjected to non-targeted metabolite profiling via liquid chromatography-mass spectrometry (LC-MS). Unsupervised multivariate analysis and Venn diagram were used to explore the relationship pattern among the house EBNs in Malaysia. The geographical distribution surrounded the swiftlet house was investigated to understand its influences on the metabolite distribution.

Results: The hierarchical clustering analysis (HCA) combined with correlation coefficient revealed the differences between the house EBNs in Malaysia with four main clusters formation. The metabolites distribution among these clusters was unique with their varied combination of geographical distribution. Cluster 1 grouped EBNs from Selangor, Melaka, Negeri Sembilan, Terengganu which geographically distributed with major oil palm field in township; Cluster 2 included Perak and Sarawak with high distribution of oil palm in higher altitude; Cluster 3 included Perlis, Kelantan, Kedah, Penang from lowland of paddy field in village mostly and Cluster 4 grouped Sabah, Pahang, Johor which are majorly distributed with undeveloped hills. The metabolites which drove each cluster formation have happened in a group instead of individual key metabolite. The major metabolites that characterised Cluster 1 were fatty acids, while the rest of the clusters were peptides and secondary metabolites.

Conclusion: The metabolite profiling conducted in this study was able to discriminate the Malaysian house EBNs based on metabolites distribution. The factor that most inferences the differences of house EBNs were the geographical distribution, in which geographical distribution affects the distribution of insect and the diet of swiftlet.
Selenium Status Is Associated With Insulin Resistance Markers in Adults: Findings From the 2013 to 2018 National Health and Nutrition Examination Survey (NHANES)
Authors: Cardoso Barbara R. , Braat Sabine , Graham Ross M.
Published: June 2021

Although literature has been consistently showing an increased risk of type 2 diabetes (T2DM) in populations with high exposure to selenium, there is a lack of information quantifying the association between diabetes-related markers and the nutritional status of selenium. Therefore, we aimed to investigate the association between blood selenium concentration and glucose markers in a representative sample of the US population, which is known to have moderate to high exposure to selenium. This cross-sectional analysis included 4,339 participants ≥18 years from the 2013 to 2018 National Health and Nutrition Examination Survey (NHANES). All participants were assessed for whole blood selenium concentration, fasting plasma insulin and glucose, HbA1c, and HOMA-IR (Homeostatic Model Assessment for Insulin Resistance). In this cohort, all participants presented with adequate selenium status [196.2 (SD: 0.9) μg/L] and 867 (15%) had diabetes mellitus. Selenium was positively associated with insulin, glucose and HOMA-IR in models adjusted for age and sex. When the models were further adjusted for smoking status, physical activity, metabolic syndrome and BMI, the associations with insulin and HOMA-IR remained but the association with glucose was no longer significant. A 10 μg/L increase in selenium was associated with 1.5% (95% CI: 0.4–2.6%) increase in insulin and 1.7% (95% CI: 0.5–2.9%) increase in HOMA-IR in fully adjusted models. There was no evidence of an association between selenium and diabetes prevalence. Our findings corroborate the notion that selenium supplementation should not be encouraged in populations with high dietary intake of selenium.
Foliar Zinc Application to Wheat May Lessen the Zinc Deficiency Burden in Rural Quzhou, China
Authors: Yu Bao-Gang , Liu Yu-Min , Chen Xiu-Xiu , Cao Wen-Qing , Ding Tong-Bin , Zou Chun-Qin
Published: June 2021

Zinc (Zn) malnutrition is a common health problem, especially in developing countries. The human health and economic benefits of the replacement of conventional flour with Zn-biofortified wheat flour in rural household diets were assessed. One hundred forty-five wheat flour samples were collected from rural households in Quzhou County. Then, field experiments were conducted on wheat at two Zn levels (0 and 0.4% ZnSO4 · 7H2O foliar application) under 16 diverse agricultural practices in Quzhou County. Foliar Zn application significantly increased the Zn concentration and bioavailability in wheat grain and flour. If rural households consumed Zn-biofortified flour instead of self-cultivated flour or flour purchased from supermarkets, 257–769 or 280–838, 0.46–1.36 million or 0.50–1.49 million disability-adjusted life years (DALYs) lost, respectively, could be saved in Quzhou County and China. Amounts of 2.3–12.0 million and 5.5–22.6 billion RMB could be obtained via Zn-biofortified flour in Quzhou County and China, respectively. The current study indicates that Zn-biofortified flour via foliar Zn application is a win-win strategy to maintain the yield and combat human Zn deficiency in rural households in China. More health and economic benefits could be obtained in rural household dependent on wheat flour purchased from supermarkets than in those dependent on self-cultivated wheat flour.
Lipidomics Identified Lyso-Phosphatidylcholine and Phosphatidylethanolamine as Potential Biomarkers for Diagnosis of Laryngeal Cancer
Authors: Yu Bo , Wang Jizhe
Published: June 2021

Background

Laryngeal cancer (LaC) remains one of the most common tumors of the respiratory tract with higher incidence in men than in women. The larynx is a small but vital organ on the neck. The dysfunction of the larynx can cause serious health problems such as hoarseness, respiratory distress, and dysphonia. Many lipids (e.g. phospholipid, cholesterol, fatty acid) have been recognized as a crucial role in tumorigenesis. However, the lipid biomarkers are lacking and the lipid molecular pathogenesis of LaC is still unclear.

Methods

This study aims to identify new LaC-related lipid biomarkers used for the diagnosis or early diagnosis of LaC and to uncover their molecular characteristics. Thus, we conducted serum and tissue nontargeted lipidomics study from LaC patients (n = 29) and normal controls (NC) (n = 36) via ultra-high performance liquid chromatography (UHPLC) coupled with high resolution mass spectrometry (HRMS). Multivariate and univariate statistics analyses were used to discriminate LaC patients from NC.

Results

As expected, a lipid panel including LPC (16:0) and PE (18:0p_20:4) was defined to distinguish the LaC patients from healthy individuals with very high diagnosis performance (area under the curve (AUC) value = 1.000, sensitivity value = 1.000, and specificity value = 1.000). In addition, the levels of Cer, CerG1, SM, PC, PC-O, PE, PI, PS, and ChE in the LaC group significantly increased as compared with the NC group. However, the levels of LPC, LPC-O, LPE, LPE-p, and DG in the LaC group significantly deceased when the one was compared with the NC group. Among significantly changed lipid species, lysophospholipids containing a palmitoyl chain or an arachidonic acid acyl chain remarkably decreased and phospholipids including a palmitoyl chain or an arachidonic acid acyl chain increased in the LaC patients.

Conclusion

Our results not only indicate that lipidomics is powerful tool to explore abnormal lipid metabolism for the laC, but suggest that lysophospholipids and phospholipids may serve as potential biomarkers for diagnosis of LaC.
Longitudinal Analysis of Circulating Tumor Cells in Colorectal Cancer Patients by a Cytological and Molecular Approach: Feasibility and Clinical Application
Authors: Hendricks Alexander , Dall Katharina , Brandt Burkhard , Geisen Reinhild , Röder Christian , Schafmayer Clemens , Becker Thomas , Hinz Sebastian , Sebens Susanne
Published: June 2021

Introduction

Liquid biopsies allowing for individualized risk stratification of cancer patients have become of high significance in individualized cancer diagnostics and treatment. The detection of circulating tumor cells (CTC) has proven to be highly relevant in risk prediction, e.g., in colorectal cancer (CRC) patients. In this study, we investigate the clinical relevance of longitudinal CTC detection over a course of follow-up after surgical resection of the tumor and correlate these findings with clinico-pathological characteristics.

Methods

In total, 49 patients with histologically proven colorectal carcinoma were recruited for this prospective study. Blood samples were analyzed for CTC presence by two methods: first by marker-dependent immunofluorescence staining combined with automated microscopy with the NYONE® cell imager and additionally, indirectly, by semi-quantitative Cytokeratin-20 (CK20) RT-qPCR. CTC quantification data were compared and correlated with the clinico-pathological parameters.

Results

Detection of CTC over a post-operative time course was feasible with both applied methods. In patients who were pre-operatively negative for CTCs with the NYONE® method or below the cut-off for relative CK20 mRNA expression after analysis by PCR, a statistically significant rise in the immediate post-operative CTC detection could be demonstrated. Further, in the cohort analyzed by PCR, we detected a lower CTC load in patients who were adjuvantly treated with chemotherapy compared to patients in the follow-up subgroup. This finding was contrary to the same patient subset analyzed with the NYONE® for CTC detection.

Conclusion

Our study investigates the occurrence of CTC in CRC patients after surgical resection of the primary tumor and during postoperative follow-up. The resection of the tumor has an impact on the CTC quantity and the longitudinal CTC analysis supports the significance of CTC as a prognostic biomarker. Future investigations with an even more extended follow-up period and larger patient cohorts will have to validate our results and may help to define an optimal longitudinal sampling scheme for liquid biopsies in the post-operative monitoring of cancer patients to enable tailored therapy concepts for precision medicine.
Molecular Markers of MDR of Chemotherapy for HSCC: Proteomic Screening With High-Throughput Liquid Chromatography-Tandem Mass Spectrometry
Authors: Shen Bin , Dong Xuelin , Yuan Bo , Zhang Zhijun
Published: June 2021

Background

Hypopharyngeal squamous cell cancer (HSCC) is a head and neck tumor with a poor prognosis. Chemotherapy lacks effectiveness because of multidrug resistance (MDR), which has increased toxic side effects. Thus, there is an urgent need to identify the molecular markers of MDR of chemotherapy for HSCC.

Methods

Fifty clinical samples of HSCC were derived from patients including 12 sensitive or resistant to chemotherapy drugs. Proteomic screening was performed using liquid chromatography-tandem mass spectrometry (LC-MS), which was based on data-independent acquisition. Molecular markers of MDR of chemotherapy in patients with HSCC were identified with clinical data and validated with ELISA.

Results

A total of 673 differentially expressed proteins were identified in HSCC samples, where 172 were upregulated and 501 were downregulated. A total of 183 differentially expressed proteins including 102 upregulated and 81 downregulated proteins, were identified by comparing cancer sensitive to chemotherapy with cancer resistant to chemotherapy. Clinical HSCC samples had significantly higher expression of FADD and significantly lower expression of RIPK1. Expressions of FADD and RIPK1 proteins were significantly lower in the chemotherapy-sensitive group. These expression differences were not correlated with clinical data. RIPK1 and FADD are involved in necroptosis and the signaling pathway of PRRs. Using ELISA, the low expression of RIPK1 and FADD was found in the patients sensitive to chemotherapy.

Conclusion

LC-MS proteomics is an effective method to identify the molecular markers of HSCC. FADD and RIPK1 can act as molecular markers of MDR of chemotherapy in patients with HSCC and may function through necroptosis and the PRR signaling pathway.
Relationship Between Ultrasound Features and Ki-67 Labeling Index of Soft Tissue Sarcoma
Authors: Wang Pingping , Li Hai , Hu Yu , Peng Xiaojing , Ye Xinhua , Xu Di , Li Ao
Published: June 2021

Objectives

To explore the relationship between ultrasound (US) features and Ki-67 labeling index (LI) of soft tissue sarcoma (STS).

Methods

Forty-six patients with 47 STS lesions, between September 2014 and April 2020, were enrolled in the study. Point-biserial correlation analysis and Spearman’s correlation analysis were utilized to examining the relationship between the US features and the Ki-67 LI of STS. The differences of US features between high and low Ki-67 proliferation groups were statistically analyzed by independent t test, Wilcoxon rank-sum test, and Fisher’s exact test. The optimal cut-off points of US features revealing significant differences were estimated by the maximum Youden index.

Results

A moderate correlation between the vascular density grade and the Ki-67 LI (ρ = 0.409, P = 0.004) was found in this study. In addition, other ultrasound features were irrelevant to the Ki-67 LI. The cut-off for differentiating low- and high-proliferation groups was grade II according to the best Youden index. The area under receiver operating characteristic (ROC) curve was 0.74 (p = 0.011) with a sensitivity of 60.6% and specificity of 78.6%.

Conclusions

Only the vascular density grade of STS had a weak positive correlation with Ki-67 LI, and might be capable of predicting the proliferation of STS. Other ultrasonographic features of STS such as shape and tumor margin have no correlation with Ki-67 LI.
Case Report: Treatment of Alectinib in NSCLC With Brain Metastasis Patient Refractory to Radiotherapy After Resistance to Crizotinib
Authors: Zhang Chunzhi
Published: June 2021

Background

Brain metastasis is the most common form of tumor recurrence after resistance to crizotinib in patients with anaplastic lymphoma kinase (ALK)-positive non-small-cell lung cancer (NSCLC). The treatment of brain metastasis in patients with ALK-positive NSCLC requires a multidisciplinary approach, including targeted therapy, chemotherapy, and radiotherapy. At present, no optimal treatment for these patients has been identified, although radiotherapy has remained a vital treatment.

Case Presentation

We experienced a patient with ALK-positive NSCLC who developed brain metastasis after crizotinib therapy. ALK rearrangement was not detected in a blood sample using next-generation sequencing. In accordance with National Comprehensive Cancer Network guidance, the patient underwent whole-brain radiotherapy. However, the number of metastatic sites unexpectedly increased. In desperation, the patient was empirically given alectinib after radiotherapy failure, and unanticipated success was achieved.

Conclusions

This case revealed some new insights. First, liquid biopsy is complementary to tissue biopsy in patients with NSCLC, mainly in those with EGFR mutation. However, ALK rearrangement should be assessed using tissue biopsy as much as possible. Second, brain metastasis of NSCLC might respond to second-generation tyrosine kinase inhibitors (TKIs), such as alectinib and ceritinib, after resistance to crizotinib regardless of the presence or absence of ALK rearrangement in liquid biopsy. Finally, combined radiotherapy and TKI therapy appears optimal in patients with brain metastasis of NSCLC after resistance to crizotinib in the absence of a definitive driver gene.
Castaneroxy A From the Leaves of Castanea sativa Inhibits Virulence in Staphylococcus aureus
Authors: Salam Akram M. , Porras Gina , Cho Young-Saeng K. , Brown Morgan M. , Risener Caitlin J. , Marquez Lewis , Lyles James T. , Bacsa John , Horswill Alexander R. , Quave Cassandra L.
Published: June 2021

Methicillin-resistant Staphylococcus aureus (MRSA) represents one of the most serious infectious disease concerns worldwide, with the CDC labeling it a “serious threat” in 2019. The current arsenal of antibiotics works by targeting bacterial growth and survival, which exerts great selective pressure for the development of resistance. The development of novel anti-infectives that inhibit quorum sensing and thus virulence in MRSA has been recurrently proposed as a promising therapeutic approach. In a follow-up of a study examining the MRSA quorum sensing inhibitory activity of extracts of Italian plants used in local traditional medicine, 224C-F2 was reported as a bioactive fraction of a Castanea sativa (European chestnut) leaf extract. The fraction demonstrated high activity in vitro and effective attenuation of MRSA pathogenicity in a mouse model of skin infection. Through further bioassay-guided fractionation using reverse-phase high performance liquid chromatography, a novel hydroperoxy cycloartane triterpenoid, castaneroxy A (1), was isolated. Its structure was established by nuclear magnetic resonance, mass spectrometry and X-ray diffraction analyses. Isomers of 1 were also detected in an adjacent fraction. In a series of assays assessing inhibition of markers of MRSA virulence, 1 exerted activities in the low micromolar range. It inhibited agr::P3 activation (IC50 = 31.72 µM), δ-toxin production (IC50 = 31.72 µM in NRS385), supernatant cytotoxicity to HaCaT human keratinocytes (IC50 = 7.93 µM in NRS385), and rabbit erythrocyte hemolytic activity (IC50 = 7.93 µM in LAC). Compound 1 did not inhibit biofilm production, and at high concentrations it exerted cytotoxicity against human keratinocytes greater than that of 224C-F2. Finally, 1 reduced dermonecrosis in a murine model of MRSA infection. The results establish 1 as a promising antivirulence candidate for development against MRSA.