bluuebunny/crossref_metadata_2025 huggingface dataset:
A Flow Cytometric Assay to Detect Functional Ganglionic Acetylcholine Receptor Antibodies by Immunomodulation in Autoimmune Autonomic Ganglionopathy
Authors: Urriola Nicolás , Spies Judith M. , Blazek Katrina , Lang Bethan , Adelstein Stephen
Published: June 2021

Autoimmune Autonomic Ganglionopathy (AAG) is an uncommon immune-mediated neurological disease that results in failure of autonomic function and is associated with autoantibodies directed against the ganglionic acetylcholine receptor (gnACHR). The antibodies are routinely detected by immunoprecipitation assays, such as radioimmunoassays (RIA), although these assays do not detect all patients with AAG and may yield false positive results. Autoantibodies against the gnACHR exert pathology by receptor modulation. Flow cytometric analysis is able to determine if this has occurred, in contrast to the assays in current use that rely on immunoprecipitation. Here, we describe the first high-throughput, non-radioactive flow cytometric assay to determine autoantibody mediated gnACHR immunomodulation. Previously identified gnACHR antibody seronegative and seropositive sera samples (RIA confirmed) were blinded and obtained from the Oxford Neuroimmunology group along with samples collected locally from patients with or without AAG. All samples were assessed for the ability to cause gnACHR immunomodulation utilizing the prototypical gnACHR expressing cell line, IMR-32. Decision limits were calculated from healthy controls, and Receiver Operating Characteristic (ROC) curves were constructed after unblinding all samples. One hundred and ninety serum samples were analyzed; all 182 expected negative samples (from healthy controls, autonomic disorders not thought to be AAG, other neurological disorders without autonomic dysfunction and patients with Systemic Lupus Erythematosus) were negative for immunomodulation (<18%), as were the RIA negative AAG and unconfirmed AAG samples. All RIA positive samples displayed significant immunomodulation. There were no false positive or negative samples. There was perfect qualitative concordance as compared to RIA, with an Area Under ROC of 1. Detection of Immunomodulation by flow cytometry for the identification of gnACHR autoantibodies offers excellent concordance with the gnACHR antibody RIA, and overcomes many of the shortcomings of immunoprecipitation assays by directly measuring the pathological effects of these autoantibodies at the cellular level. Further work is needed to determine the correlation between the degree of immunomodulation and disease severity.
Quantifying the Effects of Diver Interactions on Manta Ray Behavior at Their Aggregation Sites
Authors: Gómez-García Miguel de Jesús , Blázquez-Moreno María del Carmen , Stewart Joshua David , Leos-Barajas Vianey , Fonseca-Ponce Iliana Araceli , Zavala-Jiménez Aldo Alfonso , Fuentes Karen , Ketchum James T.
Published: June 2021

Manta rays (Mobula birostris, Mobula. cf. birostris, andMobula alfredi), the largest mobulid rays, are subjected to exploitation and overfishing in certain parts of the world. Tourism has been supported as a sustainable alternative for the conservation of the species, and a potential source of economic spillover to local populations. Nevertheless, the effects of tourism over these highly social animals remains unknown. Manta rays aggregate at three sites in Mexico: Oceanic manta rays (M. birostris)in The Revillagigedo Archipelago and Banderas Bay in the Pacific. Caribbean manta rays (M. cf. birostris) around Isla Contoy National Park in the Caribbean. We analyzed the behavior of manta rays using video data collected by local researchers and tourism operators to determine how diver behaviors and techniques (SCUBA and free diving) affect them. Diver activities were grouped into passive and active categories. We described 16 behaviors and grouped them into four behavioral states: Directional, erratic, attraction and evasion to divers. We modeled the sequence of behaviors exhibited by manta rays via first order Markov chains. Our models accounted for passive and active diver behavior when modeling the changes in manta behavior. Manta rays in Banderas Bay and Revillagigedo displayed a higher frequency of erratic behaviors than at Isla Contoy, while Banderas Bay manta rays transitioned to evasion behaviors more often. Manta rays responded similarly in both sites to active divers. At freediving sites, manta rays from Isla Contoy displayed evasion less frequently than at Banderas Bay. Changes in manta ray behavior were similar for both sites, but mantas in Banderas Bay transitioned to evasion more with active divers. The increased food availability for Isla Contoy manta rays could be the reason for the reduced response toward divers in this site. The existence of additional stressors such as both traffic in Banderas Bay could be causing the mantas in this site to respond more frequently to active divers. This study, the first of its kind in oceanic and Caribbean manta rays, highlights that regulations and the use of best practices are vital for achieving longer and less disturbing encounters for both manta rays and divers.
Measuring Sound at a Cold-Water Coral Reef to Assess the Impact of COVID-19 on Noise Pollution
Authors: De Clippele Laurence H. , Risch Denise
Published: June 2021

This study compares the noise levels at the cold-water coral Tisler reef, before and after the closure of the border between Norway and Sweden, which occurred as a direct result of the COVID-19 pandemic. The Tisler reef is a marine protected area located under a ferry “highway” that connects Norway and Sweden. Cold-water coral reefs are recognised as being important hotspots of both biodiversity and biomass, they function as breeding and nursing grounds for commercially important fish and are essential in providing ecosystem functions. Whilst studies have shown that fishery, ocean warming, and acidification threaten them, the effects of noise pollution on cold-water coral reefs remains unstudied. To study the severity of noise pollution at the Tisler reef, a long-term acoustic recorder was deployed from 29 January 2020 until 26 May 2020. From 15 March COVID-19 lockdown measures stopped passenger vessel traffic between Norway and Sweden. This study found that the overall noise levels were significantly lower after border closure, due to reduced ferry traffic, wind speeds, and sea level height. When comparing the median hourly noise levels of before vs. after border closure, this study measured a significant reduction in the 63–125 Hz 1/3 octave band noise levels of 8.94 ± 0.88 (MAD) dB during the day (07:00:00–19:59:59) and 1.94 ± 0.11 (MAD) dB during the night (20:00:00–06:59:59). Since there was no ferry traffic during the night, the drop in noise levels at night was likely driven by seasonal changes, i.e., the reduction in wind speed and sea level height when transitioning from winter to spring. Taking into account this seasonal effect, it can be deduced that the COVID-19 border closure reduced the noise levels in the 63–125 Hz 1/3 octave bands at the Tisler reef by 7.0 ± 0.99 (MAD) dB during the day. While the contribution of, and changes in biological, weather-related and geophysical sound sources remain to be assessed in more detail, understanding the extent of anthropogenic noise pollution at the Tisler cold-water coral reef is critical to guide effective management to ensure the long-term health and conservation of its ecosystem functions.
Companions and Casual Acquaintances: The Nature of Associations Among Bull Sharks at a Shark Feeding Site in Fiji
Authors: Bouveroux Thibaut , Loiseau Nicolas , Barnett Adam , Marosi Natasha D. , Brunnschweiler Juerg M.
Published: June 2021

Provisioning activities in wildlife tourism often lead to short-term animal aggregations during the feeding events. However, the presence of groups does not necessarily mean that individuals interact among each other and form social networks. At the Shark Reef Marine Reserve in Fiji, several dozen bull sharks (Carcharhinus leucas) regularly visit a site, where direct feeding is conducted during tourism driven shark dives. On 3,063 shark feeding dives between 2003 and 2016, we visually confirmed the presence of 91 individual bull sharks based on external and long-lasting identification markings. We measured the intensity of associations between pairs of individuals by calculating the Simple Ratio Index (SRI) and calculated Generalized Affiliation Indices (GAIs) to distinguish true associations between dyads from structural predictor factors. Although the resulting mean SRIs were low, ranging from 0.01 to 0.12 (SRImean= 0.06; mean SRImax= 0.21), preferred long-term companionships were observed between individuals. Avoidances were also observed within pairs of individuals during the second half of the study. The best fitting model describing the temporal association patterns of bull sharks revealed a social structure which is characterized by preferred companionships and casual acquaintances. Our results suggest that the aggregation resulting from direct feeding has served to facilitate the development of social associations.
Liver Metabolome and Proteome Response of Turbot (Scophthalmus maximus) to Lysine and Leucine in Free and Dipeptide Forms
Authors: Wei Yuliang , Li Benxiang , Xu Houguo , Liang Mengqing
Published: June 2021

Omics approaches provide more metabolic information to explain the relationship between dietary nutrition and fish growth. This study aimed to explore the metabolome and proteome response of turbot (Scophthalmus maximus) fed diets containing lysine and leucine in free and dipeptide forms by the approaches of integrated liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based metabolomics and isobaric tags for relative and absolute quantification (iTRAQ)-based proteomics. Plant protein-based diets were formulated to contain the equivalent of lysine and leucine in free amino acid [crystalline amino acid (CAA)] and synthetic Lys-Leu (Lys-Leu) forms. The metabolome and proteome profiles of the liver were screened in fish fed either the CAA diet or the Lys-Leu diet after an 8-week feeding trial. Fish fed the Lys-Leu diet showed a significantly higher final body weight and a specific growth rate compared with fish fed the CAA diet. Protein- and amino acid-related metabolic processes in the liver were identified between the Lys-Leu and CAA groups based on differential metabolites and proteins. The proteolytic enzymes and amino acid transporters from differential proteins of the liver showed that the process of protein digestion and absorption may be affected by the different forms of lysine and leucine in the feed. A mechanistic target of rapamycin complex 1 and ubiquitin proteasome pathways were identified by differential proteins, which were involved in the processes of protein synthesis and degradation in the liver. Lysine degradation, tryptophan metabolism, alanine, aspartate, and glutamate metabolism, arginine biosynthesis, arginine and proline metabolism, and glycine, serine, and threonine metabolism were identified based on differential metabolites and proteins, which showed that the metabolism of various amino acids, including lysine, had been affected by both the CAA and Lys-Leu groups. In conclusion, the data of integrated metabonomics and proteomics suggested that different forms of lysine and leucine in the feed may affect liver metabolic processes including protein digestion and absorption, protein synthesis and degradation, and amino acid metabolism. In addition, a good correlation between differential metabolites and proteins was observed in amino acid metabolism by using the approaches of integrated LC-MS/MS-based metabolomics and iTRAQ-based proteomics.
In-Situ Process and Simulation of High-Performance Piezoelectric-on-Silicon Substrate for SAW Sensor
Authors: Ma Rui , Liu Weiguo , Sun Xueping , Zhou Shun
Published: June 2021

This paper studied the manufacturing process of Piezoelectric-on-Silicon (POS) substrate which integrates 128° Y–X Lithium niobate thin film and silicon wafer using Smart-Cut technology. The blistering and exfoliation processes of the He as-implanted LN crystal under different annealing temperatures are observed by the in-situ method. Unlike the conventional polishing process, the stripping mechanism of the Lithium niobate thin film is changed by controlling annealing temperature, which can improve the surface morphology of the peeling lithium niobate thin film. We prepared the 128° Y–X POS substrate with high single-crystal Lithium niobate thin film and surface roughness of 3.91 nm through Benzocyclobutene bonding. After simulating the surface acoustic wave (SAW) characteristics of the POS substrate, the results demonstrate that the Benzocyclobutene layer not only performs as a bonding layer but also can couple more vibrations into the LN thin film. The electromechanical coupling coefficient of the POS substrate is up to 7.59% in the Rayleigh mode when hLN/λ is 0.3 and hBCB/λ is 0.1. Therefore, as a high-performance substrate material, the POS substrate has proved to be an efficient method to miniaturize and integrate the SAW sensor.
Comparative Study of the Adverse Events Associated With Adjuvant Use of Dexmedetomidine and Clonidine in Local Anesthesia
Authors: Jiang Jinjin , Shen Huasu , Zhang Jin , Wu Zhen , Shao Xian , Cui Jingjing , Zhang Bao , Ma Xiaoyu
Published: June 2021

Background: Although clonidine and dexmedetomidine are used as alpha-2 agonists to improve the quality and duration of blockade induced by local anesthetics, no study has been reported to compare their associated adverse events in local anesthesia. The aim of this study is to compare the adverse events associated with the adjuvant use of dexmedetomidine and clonidine in local anesthesia.

Methods: A comprehensive search was performed to retrieve any reported adverse event associated with adjuvant use of dexmedetomidine and clonidine in local anesthesia from published literature up to 1 July 2020. Assessment of the quality of included studies was performed by the Jadad score. A comparison of any reported adverse event was made between interventions by pooling data from studies using a direct meta-analysis technique. Dichotomous outcomes were summarized as risk ratios. The review was performed according to PRISMA guideline.

Results: From 121 articles retrieved from the search finally 14 articles including 1,120 patients had eligibility criteria for including in the meta-analysis. No significant difference was observed between bradycardia/hypotension (OR = 1.17; 95 % CI = 0.66–2.10; P = 0.580; I2 = 53.78 %, P = 0.027), nausea/vomiting (OR = 0.91; 95% CI = 0.59-1.42; P = 0.706; I2 = 0.0 %, P = 0.940) dizziness/headache (OR = 1.10; 95% CI = 0.44–2.75; P = 0.831; I2 = 0.0 %, P = 0.882) shivering (OR = 0.95 % CI = 0.50–1.66; P = 0.831; I2 = 0.0 %, P = 0.920) and dry mouth (OR = 1.00; 95 % CI = 0.50–1.96; P = 0.996; I2 = 0.0%, P = 0.900). No significant difference was observed in subgroup comparison of adverse events in the intravenous or local adjuvant use of the study drugs (p > 0.05).

Conclusion: There is no difference in adverse events associated with the intravenous or local adjuvant use of dexmedetomidine and clonidine in local anesthesia.
Understanding Immune Thrombocytopenia: Looking Out of the Box
Authors: Schifferli Alexandra , Cavalli Franco , Godeau Bertrand , Liebman Howard A. , Recher Mike , Imbach Paul , Kühne Thomas
Published: June 2021

The pathogenesis of immune thrombocytopenia (ITP) is increasingly being elucidated, and its etiology is becoming more frequently identified, leading to a diagnostic shift from primary to secondary ITP. The overlap between autoimmunity, immunodeficiency, and cancer is evident, implying more interdisciplinarity in daily care. This mini-review is based on an expert meeting on ITP organized by the Intercontinental Cooperative ITP Study Group and presents the challenges of hematologists in understanding and investigating “out of the box” concepts associated with ITP.
Exosomal MicroRNA-181a Derived From Mesenchymal Stem Cells Improves Gut Microbiota Composition, Barrier Function, and Inflammatory Status in an Experimental Colitis Model
Authors: Gu Li , Ren Feng , Fang Xianrui , Yuan Lianwen , Liu Ganglei , Wang Shalong
Published: June 2021

Background: Mesenchymal stem cell (MSC)-derived exosomes (Exos) are recently proved to be a promising candidate for ulcerative colitis (UC), but the mechanism remains unclear. We investigated the effects of MSC-derived exosomal microRNA-181a (miR-181a) on gut microbiota, immune responses, and intestinal barrier function in UC.

Methods: Human bone marrow MSC-derived Exos were extracted and identified via transmission electron microscopy (TEM), Nanoparticle Tracking Analysis (NTA), and Western blotting. Dextran sodium sulfate (DSS)-induced colitis model and lipopolysaccharide (LPS)-induced human colonic epithelial cell (HCOEPIC) model were established to determine the effect of MSC-Exos on gut microbiota, immune responses, and intestinal barrier function in vivo and in vitro. The relationship between miR-181a and UC was analyzed using the Gene Expression Omnibus (GEO) database. MSC-miR-181-inhibitor was used to reveal the role of exosomal miR-181a in DSS-induced colitis.

Results: TEM and NTA results showed that Exos of a diameter of about 100 nm with the round and oval vesicle-like structure were successfully extracted. The expressions of the CD63, CD81, and TSG101 proteins were positive in these Exos. After MSC-Exo treatment, the colon length in colitis mice increased; colon inflammatory injury decreased; TNF-α, IL-6, IL-1β, IL-17, and IL-18 levels decreased; and Claudin-1, ZO-1, and IκB levels increased. In addition, the structure of the gut microbiota in DSS-induced colitis mice was changed by MSC-Exos. MSC-Exos showed antiapoptotic effects on LPS-induced HCOEPIC. The protective effects decreased significantly by treatment with MSC-Exos interfered with miR-181a inhibitor in vivo and in vitro.

Conclusion: MSC-derived exosomal miR-181a could alleviate experimental colitis by promoting intestinal barrier function. It exerted anti-inflammatory function and affected the gut microbiota. This indicated that MSC exosomal miR-181a may exhibit potential as a disease-modifying drug for UC.
Association of Preexisting Asthma and Other Allergic Diseases With Mortality in COVID-19 Patients: A Systematic Review and Meta-Analysis
Authors: Wu Xianbo , Xu Yihua , Jin Lina , Wang Xiaoou , Zhu Haiyan , Xie Yiqiang
Published: June 2021

Background: Respiratory viruses are known to contribute to asthma exacerbations. A meta-analysis of three studies reported no association between coronavirus disease 2019 (COVID-19) mortality and preexisting asthma. This study aimed to investigate the mortality of patients with COVID-19 in relation to preexisting asthma and other allergic diseases associated with changes in respiratory function.

Methods: PubMed, Embase, and the Cochrane Library were queried for papers published up to April 9, 2021: (1) population: patients who tested positive for SARS-CoV-2 according to the WHO guidelines; (2) exposure: preexisting asthma or allergic rhinitis; (3) outcomes: mortality, ICU admission, and/or hospitalization; and (4) language: English. For studies that reported adjusted models, the most adjusted model was used for this meta-analysis; otherwise, unadjusted results were used.

Results: Twenty-four studies (1,169,441 patients) were included in this meta-analysis. Patients who died of COVID-19 were not more likely to have preexisting asthma (OR = 0.95, 95%CI: 0.78–1.15, P = 0.602; I2 = 63.5%, Pheterogeneity < 0.001). Patients with COVID-19 and admitted to the ICU (OR = 1.17, 95%CI: 0.81–1.68, P = 0.407; I2 = 91.1%, Pheterogeneity = 0.407), or hospitalized (OR = 0.91, 95%CI: 0.76–1.10, P = 0.338; I2 = 79.1%, Pheterogeneity < 0.001) were not more likely to have preexisting asthma. The results for mortality and hospitalization remained non-significant when considering the adjusted and unadjusted models separately. The results from the sensitivity analyses were consistent with the primary analyses, suggesting the robustness of our results.

Conclusion: This meta-analysis suggests that the patients who died from COVID-19, were admitted to the ICU, or hospitalized were not more likely to have asthma.
Adiponectin, Leptin, and IGF-1 Are Useful Diagnostic and Stratification Biomarkers of NAFLD
Authors: Marques Vanda , Afonso Marta B. , Bierig Nina , Duarte-Ramos Filipa , Santos-Laso Álvaro , Jimenez-Agüero Raul , Eizaguirre Emma , Bujanda Luis , Pareja Maria J. , Luís Rita , Costa Adília , Machado Mariana V. , Alonso Cristina , Arretxe Enara , Alustiza José M. , Krawczyk Marcin , Lammert Frank , Tiniakos Dina G. , Flehmig Bertram , Cortez-Pinto Helena , Banales Jesus M. , Castro Rui E. , Normann Andrea , Rodrigues Cecília M. P.
Published: June 2021

Background: Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease where liver biopsy remains the gold standard for diagnosis. Here we aimed to evaluate the role of circulating adiponectin, leptin, and insulin-like growth factor 1 (IGF-1) levels as non-invasive NAFLD biomarkers and assess their correlation with the metabolome.

Materials and Methods: Leptin, adiponectin, and IGF-1 serum levels were measured by ELISA in two independent cohorts of biopsy-proven obese NAFLD patients and healthy-liver controls (discovery: 38 NAFLD, 13 controls; validation: 194 NAFLD, 31 controls) and correlated with clinical data, histology, genetic parameters, and serum metabolomics.

Results: In both cohorts, leptin increased in NAFLD vs. controls (discovery: AUROC 0.88; validation: AUROC 0.83; p < 0.0001). The leptin levels were similar between obese and non-obese healthy controls, suggesting that obesity is not a confounding factor. In the discovery cohort, adiponectin was lower in non-alcoholic steatohepatitis (NASH) vs. non-alcoholic fatty liver (AUROC 0.87; p < 0.0001). For the validation cohort, significance was attained for homozygous for PNPLA3 allele c.444C (AUROC 0.63; p < 0.05). Combining adiponectin with specific serum lipids improved the assay performance (AUROC 0.80; p < 0.0001). For the validation cohort, IGF-1 was lower with advanced fibrosis (AUROC 0.67, p < 0.05), but combination with international normalized ratio (INR) and ferritin increased the assay performance (AUROC 0.81; p < 0.01).

Conclusion: Serum leptin discriminates NAFLD, and adiponectin combined with specific lipids stratifies NASH. IGF-1, INR, and ferritin distinguish advanced fibrosis.
Sero-Prevalence and Sero-Incidence of Antibodies to SARS-CoV-2 in Health Care Workers in Israel, Prior to Mass COVID-19 Vaccination
Authors: Muhsen Khitam , Schwaber Mitchell J. , Bishara Jihad , Kassem Eias , Atamna Alaa , Na'amnih Wasef , Goren Sophy , Bialik Anya , Mohsen Jameel , Zaide Yona , Hazan Nimrod , Ariel-Cohen Ortal , Cohen Regev , Shitrit Pnina , Marchaim Dror , Benenson Shmuel , Ben-David Debby , Rubinovitch Bina , Gotessman Tamar , Nutman Amir , Wiener-Well Yonit , Maor Yasmin , Carmeli Yehuda , Cohen Dani
Published: June 2021

Objectives: This study aims to examine the prevalence and risk factors of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sero-positivity in health care workers (HCWs), a main risk group, and assess the sero-incidence of SARS-CoV-2 infection between the first and second waves of coronavirus disease 2019 (COVID-19) in Israel.

Methods: A longitudinal study was conducted among 874 HCWs from nine hospitals. Demographics, health information, and blood samples were obtained at baseline (first wave—April–May 2020) and at follow-up (n = 373) (second wave—September–November 2020). Sero-positivity was determined based on the detection of total antibodies to the nucleocapsid antigen of SARS-CoV-2, using electro-chemiluminescence immunoassay (Elecsys® Anti-SARS-CoV-2, Roche Diagnostics, Rotkreuz, Switzerland).

Results: The sero-prevalence of SARS-CoV-2 antibodies was 1.1% [95% confidence intervals (CI) 0.6–2.1] at baseline and 8.3% (95% CI 5.9–11.6) at follow-up. The sero-conversion of SARS-CoV-2 serum antibody was 6.9% (95% CI 4.7–9.9) during the study period. The increase in SARS-CoV-2 sero-prevalence paralleled the rise in PCR-confirmed SARS-CoV-2 infections among the HCWs across the country. The likelihood of SARS-CoV-2 sero-prevalence was higher in males vs. females [odds ratio (OR) 2.52 (95% CI 1.05–6.06)] and in nurses vs. physicians [OR 4.26 (95% CI 1.08–16.77)] and was associated with being quarantined due to exposure to COVID-19 patients [OR 3.54 (95% CI 1.58–7.89)] and having a positive PCR result [OR 109.5 (95% CI 23.88–502.12)].

Conclusions: A significant increase in the risk of SARS-CoV-2 infection was found among HCWs between the first and second waves of COVID-19 in Israel. Nonetheless, the sero-prevalence of SARS-CoV-2 antibodies remains low, similar to the general population. Our findings reinforce the rigorous infection control policy, including quarantine, and utilization of personal protective equipment that should be continued together with COVID-19 immunization in HCWs and the general population.
Protein-Based Systems for Topical Antibacterial Therapy
Authors: Thapa Raj Kumar , Grønlien Krister Gjestvang , Tønnesen Hanne Hjorth
Published: June 2021

Recently, proteins are gaining attention as potential materials for antibacterial therapy. Proteins possess beneficial properties such as biocompatibility, biodegradability, low immunogenic response, ability to control drug release, and can act as protein-mimics in wound healing. Different plant- and animal-derived proteins can be developed into formulations (films, hydrogels, scaffolds, mats) for topical antibacterial therapy. The application areas for topical antibacterial therapy can be wide including bacterial infections in the skin (e.g., acne, wounds), eyelids, mouth, lips, etc. One of the major challenges of the healthcare system is chronic wound infections. Conventional treatment strategies for topical antibacterial therapy of infected wounds are inadequate, and the development of newer and optimized formulations is warranted. Therefore, this review focuses on recent advances in protein-based systems for topical antibacterial therapy in infected wounds. The opportunities and challenges of such protein-based systems along with their future prospects are discussed.
Probiotic Yeasts and Vibrio anguillarum Infection Modify the Microbiome of Zebrafish Larvae
Authors: Vargas Orlando , Gutiérrez María Soledad , Caruffo Mario , Valderrama Benjamín , Medina Daniel A. , García Katherine , Reyes-Jara Angélica , Toro Magaly , Feijóo Carmen G. , Navarrete Paola
Published: June 2021

The host microbiome plays an essential role in health and disease. Microbiome modification by pathogens or probiotics has been poorly explored especially in the case of probiotic yeasts. Next-generation sequencing currently provides the best tools for their characterization. Debaryomyces hansenii 97 (D. hansenii 97) and Yarrowia lipolytica 242 (Y. lipolytica 242) are yeasts that protect wildtype zebrafish (Danio rerio) larvae against a Vibrio anguillarum (V. anguillarum) infection, increasing their survival rate. We investigate the effect of these microorganisms on the microbiome and neutrophil response (inflammation) in zebrafish larvae line Tg(Bacmpx:GFP)i114. We postulated that preinoculation of larvae with yeasts would attenuate the intestinal neutrophil response and prevent modification of the larval microbiome induced by the pathogen. Microbiome study was performed by sequencing the V3-V4 region of the 16S rRNA gene and prediction of metabolic pathways by Piphillin in conventionally raised larvae. Survival and the neutrophil response were both evaluated in conventional and germ-free conditions. V. anguillarum infection resulted in higher neutrophil number in the intestinal area compared to non-infected larvae in both conditions. In germ-free conditions, infected larvae pre-inoculated with yeasts showed fewer neutrophil numbers than infected larvae. In both conditions, only D. hansenii 97 increased the survival of infected larvae. Beta diversity of the microbiota was modified by V. anguillarum and both yeasts, compared to non-inoculated larvae. At 3 days post-infection, V. anguillarum modified the relative abundance of 10 genera, and pre-inoculation with D. hansenii 97 and Y. lipolytica 242 prevented the modification of 5 and 6 of these genera, respectively. Both yeasts prevent the increase of Ensifer and Vogesella identified as negative predictors for larval survival (accounting for 40 and 27 of the variance, respectively). In addition, yeast pre-inoculation prevents changes in some metabolic pathways altered by V. anguillarum’s infection. These results suggest that both yeasts and V. anguillarum can shape the larval microbiota configuration in the early developmental stage of D. rerio. Moreover, modulation of key taxa or metabolic pathways of the larval microbiome by yeasts can be associated with the survival of infected larvae. This study contributes to the understanding of yeast–pathogen–microbiome interactions, although further studies are needed to elucidate the mechanisms involved.
pH Changes Have a Profound Effect on Gene Expression, Hydrolytic Enzyme Production, and Dimorphism in Saccharomycopsis fibuligera
Authors: Farh Mohamed El-Agamy , Abdellaoui Najib , Seo Jeong-Ah
Published: June 2021

Saccharomycopsis fibuligera is an amylolytic yeast that plays an important role within nuruk (a traditional Korean fermentation starter) used for the production of makgeolli (Korean rice wine), which is characterized by high acidity. However, the effect of pH change (neutral to acidic) on the yeast cell to hyphal transition and carbohydrate-hydrolyzing enzyme activities for S. fibuligera has not been investigated yet. In this study, S. fibuligera strains were cultured under the different pH conditions, and the effect on the enzyme production and gene expression were investigated. An acidic pH induced a hyphal transition from yeast cell of S. fibuligera KPH12 and the hybrid strain KJJ81. In addition, both strains showed a gradual decrease in the ability to degrade starch and cellulose as the pH went down. Furthermore, a transcriptome analysis demonstrated that the pH decline caused global expression changes in genes, which were classified into five clusters. Among the differentially expressed genes (DEGs) under acidic pH, the downregulated genes were involved in protein synthesis, carbon metabolism, and RIM101 and cAMP-PKA signaling transduction pathways for the yeast-hyphal transition. A decrease in pH induced a dimorphic lifestyle switch from yeast cell formation to hyphal growth in S. fibuligera and caused a decrease in carbohydrate hydrolyzing enzyme production, as well as marked changes in the expression of genes related to enzyme production and pH adaptation. This study will help to elucidate the mechanism of adaptation of S. fibuligera to acidification that occur during the fermentation process of makgeolli using nuruk.
The Role of Human Papilloma Virus in Dictating Outcomes in Head and Neck Squamous Cell Carcinoma
Authors: Brennan Shane , Baird Anne-Marie , O’Regan Esther , Sheils Orla
Published: June 2021

The Human Papilloma Virus (HPV) is an oncogenic virus which is associated with the development of head and neck squamous cell carcinoma (HNSCC), predominantly within the oropharynx. Approximately 25% of oropharyngeal squamous cell carcinoma (OPSCC) cases worldwide are attributable to HPV infection, with an estimated 65% in the United States. Transmission is via exposure during sexual contact, with distinctive anatomical features of the tonsils providing this organ with a predilection for infection by HPV. No premalignant lesion is identifiable on clinical examination, thus no comparative histological features to denote the stages of carcinogenesis for HPV driven HNSCC are identifiable. This is in contrast to HPV-driven cervical carcinoma, making screening a challenge for the head and neck region. However, HPV proffers a favorable prognosis in the head and neck region, with better overall survival rates in contrast to its HPV negative counterparts. This has resulted in extensive research into de-intensifying therapies aiming to minimize the morbidity induced by standard concurrent chemo-radiotherapy without compromising efficacy. Despite the favorable prognosis, cases of recurrence and/or metastasis of HPV positive HNSCC do occur, and are linked with poor outcomes. HPV 16 is the most frequent genotype identified in HNSCC, yet there is limited research to date studying the impact of other HPV genotype with respect to overall survival. A similar situation pertains to genetic aberrations associated in those with HPV positive HNSCC who recur, with only four published studies to date. Somatic mutations in TSC2, BRIP1, NBN, TACC3, NFE2l2, STK11, HRAS, PIK3R1, TP63, and FAT1 have been identified in recurrent HPV positive OPSCC. Finding alternative therapeutic strategies for this young cohort may depend on upfront identification of HPV genotypes and mutations which are linked with worse outcomes, thus ensuring appropriate stratification of treatment regimens.
Association Between Visceral Fat and Brain Cortical Thickness in the Elderly: A Neuroimaging Study
Authors: Cho Jaelim , Seo Seongho , Kim Woo-Ram , Kim Changsoo , Noh Young
Published: June 2021

Background

Despite emerging evidence suggesting that visceral fat may play a major role in obesity-induced neurodegeneration, little evidence exists on the association between visceral fat and brain cortical thickness in the elderly.

Purpose

We aimed to examine the association between abdominal fat and brain cortical thickness in a Korean elderly population.

Methods

This cross-sectional study included elderly individuals without dementia (n= 316). Areas of visceral fat and subcutaneous fat (cm2) were estimated from computed tomography scans. Regional cortical thicknesses (mm) were obtained by analyzing brain magnetic resonance images. Given the inverted U-shaped relationship between visceral fat area and global cortical thickness (examined using a generalized additive model), visceral fat area was categorized into quintiles, with the middle quintile being the reference group. A generalized linear model was built to explore brain regions associated with visceral fat. The same approach was used for subcutaneous fat.

Results

The mean (standard deviation) age was 67.6 (5.0) years. The highest quintile (vs. the middle quintile) group of visceral fat area had reduced cortical thicknesses in the global [β = –0.04 mm, standard error (SE) = 0.02 mm,p= 0.004], parietal (β = –0.04 mm,SE= 0.02 mm,p= 0.01), temporal (β = –0.05 mm,SE= 0.02 mm,p= 0.002), cingulate (β = –0.06 mm,SE= 0.02 mm,p= 0.01), and insula lobes (β = –0.06 mm,SE= 0.03 mm,p= 0.02). None of the regional cortical thicknesses significantly differed between the highest and the middle quintile groups of subcutaneous fat area.

Conclusion

The findings suggest that a high level of visceral fat, but not subcutaneous fat, is associated with a reduced cortical thickness in the elderly.
Evaluation of Peripheral Immune Activation in Amyotrophic Lateral Sclerosis
Authors: Wang Mengli , Liu Zhen , Du Juan , Yuan Yanchun , Jiao Bin , Zhang Xuewei , Hou Xuan , Shen Lu , Guo Jifeng , Jiang Hong , Xia Kun , Tang Jianguang , Zhang Ruxu , Tang Beisha , Wang Junling
Published: June 2021

Accumulating evidence has revealed that immunity plays an important role in amyotrophic lateral sclerosis (ALS) progression. However, the results regarding the serum levels of immunoglobulin and complement are inconsistent in patients with ALS. Although immune dysfunctions have also been reported in patients with other neurodegenerative diseases, few studies have explored whether immune dysfunction in ALS is similar to that in other neurodegenerative diseases. Therefore, we performed this study to address these gaps. In the present study, serum levels of immunoglobulin and complement were measured in 245 patients with ALS, 65 patients with multiple system atrophy (MSA), 60 patients with Parkinson's disease (PD), and 82 healthy controls (HCs). Multiple comparisons revealed that no significant differences existed between patients with ALS and other neurodegenerative diseases in immunoglobulin and complement levels. Meta-analysis based on data from our cohort and eight published articles was performed to evaluate the serum immunoglobulin and complement between patients with ALS and HCs. The pooled results showed that patients with ALS had higher C4 levels than HCs. In addition, we found that the IgG levels were lower in early-onset ALS patients than in late-onset ALS patients and HCs, and the correlations between age at onset of ALS and IgG or IgA levels were significant positive. In conclusion, our data supplement existing literature on understanding the role of peripheral immunity in ALS.
Romanian GPs Involvement in Caring for the Mental Health Problems of the Elderly Population: A Cross-Sectional Study
Authors: Sfetcu Raluca , Toma Daciana , Tudose Catalina , Vladescu Cristian
Published: June 2021

The mental health of the elderly is a matter of increased concern in the context of an aging population since currently only a small fraction of this population is receiving adequate care. The provision of treatment in primary care by the General Practitioners (GPs) has been proposed for over a decade as a potential solution, as services offered by GPs are more accessible, less susceptible to stigma, and have a more comprehensive view of the other health care problems that the elderly might suffer from. In this study, we explored the perception of Romanian GPs regarding their practice and roles in caring for the mental health of the elderly as well as the willingness to increase their future involvement in the management of dementia and other mental health problems. Data was collected via an online questionnaire structured on four dimensions: (1) GPs' sociodemographic profile and practice characteristics, (2) GPs assessment of the services available for elderly with mental health problems, (3) GPs current involvement in mental health care for different categories of problems, and (4) factors that might influence the future involvement of GPs in providing care for elderly with mental health problems. The survey was sent via the member mailing lists of the National Society for Family Medicine. Results show that GPs are currently limited by prescribing possibilities, available resources and knowledge in the area, but they are willing to expand their role in the areas of early recognition and prevention of mental health problems as well as providing disease management and collaborative care. An improved communication with mental health care professionals, a better access to resources and having more financial incentives are the three most important categories for GPs to increase their involvement. In conclusion, increasing the access to personal and professional resources and setting up functional communication channels with specialized mental health care could motivate GPs to provide timely mental health support to elderly patients.
Hierarchical Encoder-Decoder With Soft Label-Decomposition for Mitochondria Segmentation in EM Images
Authors: Luo Zhengrong , Wang Ye , Liu Shikun , Peng Jialin
Published: June 2021

Semantic segmentation of mitochondria from electron microscopy (EM) images is an essential step to obtain reliable morphological statistics about mitochondria. However, automatically delineating plenty of mitochondria of varied shapes from complex backgrounds with sufficient accuracy is challenging. To address these challenges, we develop a hierarchical encoder-decoder network (HED-Net), which has a three-level nested U-shape architecture to capture rich contextual information. Given the irregular shape of mitochondria, we introduce a novel soft label-decomposition strategy to exploit shape knowledge in manual labels. Rather than simply using the ground truth label maps as the unique supervision in the model training, we introduce additional subcategory-aware supervision by softly decomposing each manual label map into two complementary label maps according to mitochondria's ovality. The three label maps are integrated with our HED-Net to supervise the model training. While the original label map guides the network to segment all the mitochondria of varied shapes, the auxiliary label maps guide the network to segment subcategories of mitochondria of circular shape and elliptic shape, respectively, which are much more manageable tasks. Extensive experiments on two public benchmarks show that our HED-Net performs favorably against state-of-the-art methods.
Evaluation of Neuropathological Features in the SOD1-G93A Low Copy Number Transgenic Mouse Model of Amyotrophic Lateral Sclerosis
Authors: Molnar-Kasza Agnes , Hinteregger Barbara , Neddens Joerg , Rabl Roland , Flunkert Stefanie , Hutter-Paier Birgit
Published: June 2021

Amyotrophic lateral sclerosis (ALS) still depicts an incurable and devastating disease. Drug development efforts are mostly based on superoxide dismutase 1 gene (SOD1)-G93A mice that present a very strong and early phenotype, allowing only a short time window for intervention. An alternative mouse model is available, that is based on the same founder line but has a reduced SOD1-G93A copy number, resulting in a weaker and delayed phenotype. To be able to use these SOD1-G93A/low mice for drug testing, we performed a characterization of ALS-typical pathologies. All analyses were performed compared to non-transgenic (ntg) littermates of the same sex and age. In vivo analysis of SOD1-G93A/low mice was performed by weekly body weight measurements, analysis of the survival rate, and measurement of the muscle strength of 24–30 weeks old female and male SOD1-G93A/low mice. Immunofluorescent labeling of SOD1, glial fibrillary acidic protein (GFAP), and ionized calcium-binding adaptor molecule 1 (Iba1) protein was performed in the cervical, thoracic, and lumbar ventral horn of the spinal cord of 24–30 weeks old male and female SOD1-G93A/low mice. The musculus gastrocnemius of male SOD1-G93A/low mice was labeled with fluorophore-conjugated α-bungarotoxin and antibodies against phosphorylated neurofilaments. Fluorescent labeling was detected and quantified by macro-based image analysis. Although SOD1 protein levels were highly increased in both sexes and all age groups, levels strongly peaked in 30 weeks old male SOD1-G93A/low mice. Astrocytosis and activated microglia in the spinal cord ventral horn and phosphorylated neurofilaments in the motor unit of the musculus gastrocnemius progressively increased, while muscle strength progressively decreased in male SOD1-G93A/low mice. In female SOD1-G93A/low mice, only activated microglia increased progressively, while muscle strength was constantly reduced starting at 26 weeks. These differences result in a shorter survival time of male SOD1-G93A/low mice of about 3 weeks compared to female animals. The results suggest that male SOD1-G93A/low mice present a stronger pathology and are, therefore, better suitable to evaluate the efficacy of new drugs against ALS as most pathological features are developing progressively paralleled by a survival time that allows treatment to start before symptom onset.
Deep Learning for Differentiating Benign From Malignant Parotid Lesions on MR Images
Authors: Xia Xianwu , Feng Bin , Wang Jiazhou , Hua Qianjin , Yang Yide , Sheng Liang , Mou Yonghua , Hu Weigang
Published: June 2021

Purpose/Objectives(s)

Salivary gland tumors are a rare, histologically heterogeneous group of tumors. The distinction between malignant and benign tumors of the parotid gland is clinically important. This study aims to develop and evaluate a deep-learning network for diagnosing parotid gland tumors via the deep learning of MR images.

Materials/Methods

Two hundred thirty-three patients with parotid gland tumors were enrolled in this study. Histology results were available for all tumors. All patients underwent MRI scans, including T1-weighted, CE-T1-weighted and T2-weighted imaging series. The parotid glands and tumors were segmented on all three MR image series by a radiologist with 10 years of clinical experience. A total of 3791 parotid gland region images were cropped from the MR images. A label (pleomorphic adenoma and Warthin tumor, malignant tumor or free of tumor), which was based on histology results, was assigned to each image. To train the deep-learning model, these data were randomly divided into a training dataset (90%, comprising 3035 MR images from 212 patients: 714 pleomorphic adenoma images, 558 Warthin tumor images, 861 malignant tumor images, and 902 images free of tumor) and a validation dataset (10%, comprising 275 images from 21 patients: 57 pleomorphic adenoma images, 36 Warthin tumor images, 93 malignant tumor images, and 89 images free of tumor). A modified ResNet model was developed to classify these images. The input images were resized to 224x224 pixels, including four channels (T1-weighted tumor images only, T2-weighted tumor images only, CE-T1-weighted tumor images only and parotid gland images). Random image flipping and contrast adjustment were used for data enhancement. The model was trained for 1200 epochs with a learning rate of 1e-6, and the Adam optimizer was implemented. It took approximately 2 hours to complete the whole training procedure. The whole program was developed with PyTorch (version 1.2).

Results

The model accuracy with the training dataset was 92.94% (95% CI [0.91, 0.93]). The micro-AUC was 0.98. The experimental results showed that the accuracy of the final algorithm in the diagnosis and staging of parotid cancer was 82.18% (95% CI [0.77, 0.86]). The micro-AUC was 0.93.

Conclusion

The proposed model may be used to assist clinicians in the diagnosis of parotid tumors. However, future larger-scale multicenter studies are required for full validation.
Tumor Immune Microenvironment and Genetic Alterations in Mesothelioma
Authors: Hiltbrunner Stefanie , Mannarino Laura , Kirschner Michaela B. , Opitz Isabelle , Rigutto Angelica , Laure Alexander , Lia Michela , Nozza Paolo , Maconi Antonio , Marchini Sergio , D’Incalci Maurizio , Curioni-Fontecedro Alessandra , Grosso Federica
Published: June 2021

Malignant pleural mesothelioma (MPM) is a rare and fatal disease of the pleural lining. Up to 80% of the MPM cases are linked to asbestos exposure. Even though its use has been banned in the industrialized countries, the cases continue to increase. MPM is a lethal cancer, with very little survival improvements in the last years, mirroring very limited therapeutic advances. Platinum-based chemotherapy in combination with pemetrexed and surgery are the standard of care, but prognosis is still unacceptably poor with median overall survival of approximately 12 months. The genomic landscape of MPM has been widely characterized showing a low mutational burden and the impairment of tumor suppressor genes. Among them, BAP1 and BLM are present as a germline inactivation in a small subset of patients and increases predisposition to tumorigenesis. Other studies have demonstrated a high frequency of mutations in DNA repair genes. Many therapy approaches targeting these alterations have emerged and are under evaluation in the clinic. High-throughput technologies have allowed the detection of more complex molecular events, like chromotripsis and revealed different transcriptional programs for each histological subtype. Transcriptional analysis has also paved the way to the study of tumor-infiltrating cells, thus shedding lights on the crosstalk between tumor cells and the microenvironment. The tumor microenvironment of MPM is indeed crucial for the pathogenesis and outcome of this disease; it is characterized by an inflammatory response to asbestos exposure, involving a variety of chemokines and suppressive immune cells such as M2-like macrophages and regulatory T cells. Another important feature of MPM is the dysregulation of microRNA expression, being frequently linked to cancer development and drug resistance. This review will give a detailed overview of all the above mentioned features of MPM in order to improve the understanding of this disease and the development of new therapeutic strategies.