bluuebunny/crossref_metadata_2025 huggingface dataset:
Endothelin B Receptor Immunodynamics in Pulmonary Arterial Hypertension
Authors: Tabeling Christoph , González Calera Carla R. , Lienau Jasmin , Höppner Jakob , Tschernig Thomas , Kershaw Olivia , Gutbier Birgitt , Naujoks Jan , Herbert Julia , Opitz Bastian , Gruber Achim D. , Hocher Berthold , Suttorp Norbert , Heidecke Harald , Burmester Gerd-R. , Riemekasten Gabriela , Siegert Elise , Kuebler Wolfgang M. , Witzenrath Martin
Published: June 2022

Introduction

Inflammation is a major pathological feature of pulmonary arterial hypertension (PAH), particularly in the context of inflammatory conditions such as systemic sclerosis (SSc). The endothelin system and anti-endothelin A receptor (ETA) autoantibodies have been implicated in the pathogenesis of PAH, and endothelin receptor antagonists are routinely used treatments for PAH. However, immunological functions of the endothelin B receptor (ETB) remain obscure.

Methods

Serum levels of anti-ETB receptor autoantibodies were quantified in healthy donors and SSc patients with or without PAH. Age-dependent effects of overexpression of prepro-endothelin-1 or ETB deficiency on pulmonary inflammation and the cardiovascular system were studied in mice. Rescued ETB-deficient mice (ETB-/-) were used to prevent congenital Hirschsprung disease. The effects of pulmonary T-helper type 2 (Th2) inflammation on PAH-associated pathologies were analyzed in ETB-/- mice. Pulmonary vascular hemodynamics were investigated in isolated perfused mouse lungs. Hearts were assessed for right ventricular hypertrophy. Pulmonary inflammation and collagen deposition were assessed via lung microscopy and bronchoalveolar lavage fluid analyses.

Results

Anti-ETB autoantibody levels were elevated in patients with PAH secondary to SSc. Both overexpression of prepro-endothelin-1 and rescued ETB deficiency led to pulmonary hypertension, pulmonary vascular hyperresponsiveness, and right ventricular hypertrophy with accompanying lymphocytic alveolitis. Marked perivascular lymphocytic infiltrates were exclusively found in ETB-/- mice. Following induction of pulmonary Th2 inflammation, PAH-associated pathologies and perivascular collagen deposition were aggravated in ETB-/- mice.

Conclusion

This study provides evidence for an anti-inflammatory role of ETB. ETB seems to have protective effects on Th2-evoked pathologies of the cardiovascular system. Anti-ETB autoantibodies may modulate ETB-mediated immune homeostasis.
Increased Coastal Nutrient Loading Enhances Reproductive Intensity of Zostera marina: Implications for Seagrass Meadow Resilience
Authors: Suonan Zhaxi , Kim Seung Hyeon , Qin Le-Zheng , Kim Hyegwang , Zhang Fei , Lee Kun-Seop
Published: June 2022

Because sexual reproduction is essential for the establishment and persistence of seagrass meadows, flowering intensity is an important trait that influences the resilience and stability of seagrass populations. Although the effects of excessive coastal nutrient loading on seagrass vegetative growth have been extensively documented, the effects on seagrass reproductive phenology and intensity remain unclear. To examine the reproductive responses of seagrass populations to increased coastal nutrient loading, the flowering phenology and intensity of Zostera marina were compared between sites with high-nutrient, low-light conditions (Deukryang Bay and Dongdae Bay) and low-nutrient, high-light conditions (Koje Bay) on the southern coast of Korea. Nutrient contents of the above- and below-ground tissues of Z. marina reflected in situ nutrient and light availability at the study sites. Reproductive shoot density and biomass, as well as flowering frequency and reproductive effort, were much higher (1.5–4.6-fold) at the high-nutrient, low-light study sites of Deukryang Bay and Dongdae Bay than at the low-nutrient, high-light site of Koje Bay. Consequently, potential seed production was higher in Deukryang Bay and Dongdae Bay than in Koje Bay. Chronic high-nutrient and low-light conditions significantly increased the reproductive intensity of Z. marina, supporting the persistence and resilience of Z. marina populations. The results of this study could provide insights into the conservation and management of seagrass meadows under increased coastal nutrient loading.
Benthic and Pelagic Contributions to Primary Production: Experimental Insights From the Gulf of Trieste (Northern Adriatic Sea)
Authors: Cibic Tamara , Baldassarre Laura , Cerino Federica , Comici Cinzia , Fornasaro Daniela , Kralj Martina , Giani Michele
Published: June 2022

Although the 14C-method remains one of the most sensitive measures of primary production in marine ecosystems, few data from coastal sublittoral areas are available. We applied an integrated approach to quantify the benthic (PPs) and pelagic (PPw) contributions to total primary production (PPt) in a 17-m deep coastal site. From March 2015 to March 2019, we carried out 16 in situ experiments on a seasonal basis, at the LTER site C1, whereas benthic rates were estimated in the laboratory. To relate PP to seawater physical features and to the water column stability, the Brunt-Väisälä frequency was calculated. We further related our PP rates to the abundance, biomass, main taxonomic groups and diversity of eukaryotic phytoplankton and microphytobenthos (MPB). In November 2018, the maximum PPw (6.71 ± 0.82 µgC L-1 h-1) was estimated at the surface layer, in correspondence to the highest value of dinoflagellates biomass (29.35 µgC L-1), on the account of small (<20 µm) naked and thecate forms. PPi, integrated over the water column, displayed the highest values in July 2017 and July 2018. In sediments, negative PPs values were estimated in late autumn/winter, when minima of MPB abundance occurred. The highest rates were displayed in January 2018 and October 2016 (28.50 and 17.55 mgC m-2 h-1), due to the presence of dominant diatoms Paralia sulcata and Nitzschia sigma var. sigmatella, respectively. The PPs contribution to PPt was negligible (<2%) in 6 out of 16 experiments, with a mean value of 11.3% (excluding negative PPs values) over the study period, while it reached up to 43% in January 2018. The principal component analyses revealed that nutrients availability affected the seasonal development of pelagic and benthic phototrophs and primary production more than the physical variables, except for the surface layer of the water column where temperature and salinity were the main drivers. Our results add on the limited database on primary production in sublittoral areas and represent one of the few attempts, on a global scale, of integrating pelagic and benthic primary production using the 14C method to quantify the overall ecosystem productivity.
Temporal Variation of Summer Hypoxia off Changjiang Estuary During 1997–2014 and Its Association With ENSO
Authors: Ma Xiao , Liu Anqi , Zhao Qiang , Wang Bin , Tian Di , Meng Qicheng , Zeng Dingyong , Li Jia , Huang Daji , Zhou Feng
Published: June 2022

Temporal variation of summer hypoxia, along a repeated hydrographic section from the Changjiang River mouth to Cheju Island, is investigated using cruise data during 1997–2014. The climatological mean of dissolved oxygen (DO) presents a “dual-core structure” below the oxycline, associated with two hypoxic centers near the river mouth and offshore of the Changjiang Bank, respectively. The DO value is highly variable from year to year, with dramatic interannual variation but an insignificant linear trend during 1997–2014. Composite analysis indicates that the interannual variation of DO and associated hypoxia events are strongly affected by the El Niño–Southern Oscillation (ENSO) cycle. In the El Niño decaying summer, large river discharge and relatively strong stratification, combined with calm climatic and hydrographic conditions, enhance DO depletion off the Changjiang Estuary. It is likely that El Niño is not a prerequisite for hypoxia occurrences. However, El Niño did enhance bottom hypoxia probability off the Changjiang Estuary during 1997–2014.
Antioxidant, Transcriptomic and Metabonomic Analysis of Hepatopancreatic Stress Resistance in Exopalaemon carinicauda Following Astaxanthin Feeding
Authors: Li Wenyang , Wang Jiajia , Li Jitao , Liu Ping , Li Jian , Zhao Fazhen
Published: June 2022

Astaxanthin (Axn) is a xanthophyll carotenoid that has previously been shown to suppress hepatic inflammation, reduce oxidative liver damage, and improve metabolic profiles. Exopalaemon carinicauda (E. carinicauda) is an economically important fishery species in China that has been found to exhibit increased body weight following Axn feeding as compared to a standard diet. In this study, dietary Axn can significantly decreased MDA content, T-AOC and significantly increased SOD, GSH and CAT activities in shrimp hepatopancreas. Moreover, transcriptome and metabolome of E. carinicauda after Axn feeding were investigated to identify the mechanism of the effect of Axn on E. carinicauda. The transcriptomic data revealed that a total 99 different expression genes (DEGs) were identified between the Axn and control groups, of which 47 and 52 were upregulated and downregulated, respectively. DEGs of E. carinicauda such as catherpsin, actin and PARP after Axn feeding were associated with apoptosis and immune system. The metabolomic analysis revealed that A total of 73 different expression metabolites (DEMs) were identified in both metabolites, including 30 downregulated metabolites and 43 upregulated metabolites. And Axn participate in metabolism processes in hepatopancreas of E. carinicauda, including the TCA cycle, amino acid metabolism and lipid metabolism. The multiple comparative analysis implicated that Axn can improve the antioxidant capacity of hepatopancreas and the energy supply of hepatopancreas mitochondria, and then improve the ability of anti-apoptosis. Collectively, all these results will greatly provide new insights into the molecular mechanisms underlying tolerance of adverse environment in E. carinicauda.
Screening New Blood Indicators for Non-alcoholic Fatty Liver Disease (NAFLD) Diagnosis of Chinese Based on Machine Learning
Authors: Wang Cheng , Yan Junbin , Zhang Shuo , Xie Yiwen , Nie Yunmeng , Chen Zhiyun , Xu Sumei
Published: June 2022

Background

The prevalence of NAFLD is increasing annually. The early diagnosis and control are crucial for the disease. Currently, metabolic indicators are always used clinically as an auxiliary diagnosis of NAFLD. However, the prevalence of NAFLD is not only increased in obese/metabolic-disordered populations. NAFLD patients with thin body are also increasing. Only using metabolic indicators to assist in the diagnosis of NAFLD may have some deficiencies. Continue to develop more clinical auxiliary diagnostic indicators is pressing.

Methods

Machine learning methods are applied to capture risk factors for NAFLD in 365 adults from Zhejiang Province. Predictive models are constructed for NAFLD using fibrinolytic indicators and metabolic indicators as predictors respectively. Then the predictive effects are compared; ELISA kits were used to detect the blood indicators of non-NAFLD and NAFLD patients and compare the differences.

Results

The prediction accuracy for NAFLD based on fibrinolytic indicators [Tissue Plasminogen Activator (TPA), Plasminogen Activator Inhibitor-1 (PAI-1)] is higher than that based on metabolic indicators. TPA and PAI-1 are more suitable than metabolic indicators to be selected to predict NAFLD.

Conclusions

The fibrinolytic indicators have a stronger association with NAFLD than metabolic indicators. We should attach more importance to TPA and PAI-1, in addition to TC, HDL-C, LDL-C, and ALT/AST, when conducting blood tests to assess NAFLD.
Cardiac-Related Lesions in Newly Diagnosed Patients With Acute Leukemia: A Chinese Population-Based Real-World Study
Authors: Xiao Wei , Ma Linlu , Shang Yufeng , Yang Fuwei , Tan Yuxin , Chen Guopeng , Wu Jinxian , Liang Yuxing , Rouzi Tuerxunayi , Wang Qian , Zhang Nan , Zhou Fuling
Published: June 2022

The relationship between newly diagnosed acute leukemia (AL) and heart-related lesions remains unclear. This study aimed to investigate baseline cardiac function and risk of cardiovascular diseases (CVDs) in patients with new-onset AL, and provide data on cardiac management strategies for patients with AL. We retrospectively collected data on baseline characteristics, echocardiography, and biochemical blood indicators (e.g., myocardial enzymes) from 408 patients, 200 with newly diagnosed AL, 103 with coronary artery disease (CAD), and 105 controls from January 1, 2015 to August 31, 2019. The creatine kinase isoenzyme myocardial band, lactate dehydrogenase, highly sensitive troponin-I, and B-type natriuretic peptide levels and left ventricular internal diameter (LVID) were significantly higher in patients with newly diagnosed AL than in the control group. The degree of cardiac damage was lower in newly diagnosed AL patients than in CAD patients. The best predictor of heart damage was LVID (AUC [area under the curve] = 0.709; 95% CI [confidence interval]: 0.637–0.781; p < 0.001), and independent prognostic risk factors were age and ejection fraction (HR [hazard ratio] = 1.636; 95% CI: 1.039–2.575; p = 0.033). The ratio of leukemia blasts among patients with AL was positively correlated with cardiac damage. Our data indicated that newly diagnosed AL patients had certain myocardial damage before treatment. Clinicians need to pay attention to these manifestations, which may be related to the prognosis.
Assessment Methods in Medical Ultrasound Education
Authors: Höhne Elena , Recker Florian , Dietrich Christoph Frank , Schäfer Valentin Sebastian
Published: June 2022

Medical schools are increasingly incorporating ultrasound into undergraduate medical education. The global integration of ultrasound into teaching curricula and physical examination necessitates a strict evaluation of the technology's benefit and the reporting of results. Course structures and assessment instruments vary and there are no national or worldwide standards yet. This systematic literature review aims to provide an up-to-date overview of the various formats for assessing ultrasound skills. The key questions were framed in the PICO format (Population, Intervention, Comparator, and Outcome). A review of literature using Embase, PubMed, Medline, Cochrane and Google Scholar was performed up to May 2021, while keywords were predetermined by the authors. Inclusion criteria were as follows: prospective as well as retrospective studies, observational or intervention studies, and studies outlining how medical students learn ultrasound. In this study, 101 articles from the literature search matched the inclusion criteria and were investigated. The most frequently used methods were objective structured clinical examinations (OSCE), multiple choice questions, and self-assessments via questionnaires while frequently more than one assessment method was applied. Determining which assessment method or combination is ideal to measure ultrasound competency remains a difficult task for the future, as does the development of an equitable education approach leading to reduced heterogeneity in curriculum design and students attaining equivalent skills.
Rapid and Sensitive Qualitative Duoplex Real-Time PCR Method for Discriminatory and Confirmatory Diagnosis of HTLV-1 and HTLV-2 Infections: Brazilian Multicentric Study
Authors: Rocha-Junior Mauricio Cristiano , Rodrigues Evandra Strazza , Slavov Svetoslav Nanev , Assone Tatiane , Pedreschi Maíra , de La Roque Debora Glenda Lima , Sousa Maisa , Olavarria Viviana , Galvão-Castro Bernardo , Fonseca Benedito Antonio Lopes da , Penalva de Oliveira Augusto César , Smid Jerusa , Takayanagui Oswaldo Massaiti , Casseb Jorge , Covas Dimas Tadeu , Kashima Simone
Published: June 2022

Human T cell lymphotropic virus (HTLV) is the caustive agent of two main conditions i. e., the HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and the adult T-cell leukemia/lymphoma (ATLL). HTLV diagnosis is based on serological and molecular approaches; however, an accurate and validated method is still needed. The objective of this study was to establish a rapid and sensitive molecular test to confirm and discriminate HTLV 1/2 types. The test validation was performed as a multicentric study involving HTLV confirmation centers throughout Brazil. Proviral DNA was extracted from whole blood and the amplification was performed using in-house designed primer and probe sets targeting the pol genomic region. An internal control to validate the extraction and amplification was also included. The limit of detection (LoD) of the assay was four copies/reaction for HTLV-1 and 10.9 copies/reaction for HTLV-2. The diagnostic sensitivity of the platform was 94.6% for HTLV-1, 78.6% for HTLV-2, and the specificity was 100% for both viruses. Cross-reactions of the test with human viruses including HAV, HBV, HCV, HIV-1/2, and parvovirus B19 were not observed. During the multicentric validation, the test was used to screen a total of 692 blood samples obtained from previously confirmed HTLV-positive individuals. From these, 91.1% tested positive being concordant with the previously obtained results. In conclusion, our duoplex-RT-PCR-HTLV1 /2 presented adequate efficiency for HTLV-1/2 differentiation showing high sensitivity and specificity. Therefore, it can be a suitable tool for confirmation of suspected and inconclusive HTLV cases, prenatal and pre-transplant diagnosis, in Brazil and in other countries HTLV-endemic countries.
The Impact of Seropositivity on Systemic Bone Loss in Rheumatoid Arthritis—A 3-Year Interim Analysis of a Longitudinal Observational Cohort Study
Authors: Yu Shan-Fu , Chen Jia-Feng , Chen Ying-Chou , Wang Yu-Wei , Hsu Chung-Yuan , Lai Han-Ming , He Hsiao-Ru , Ko Chi-Hua , Chiu Wen-Chan , Cheng Tien-Tsai
Published: June 2022

Objective

To explore the impact of seropositivity on systemic bone loss in rheumatoid arthritis (RA).

Methods

We conducted an interim analysis of the RA registry. Patients were examined with dual-energy X-ray absorptiometry at baseline and again 3 years later. Participants were grouped into seropositive (SPRA) and seronegative (SNRA) based on the presence or absence of rheumatoid factor (RF) and/or anti-cyclic citrullinated peptide antibodies (ACPA). After matching (1:2) for age and sex, SNRA and SPRA patients were divided into groups A and B. Each matched group (A or B) was further subdivided according to the number of antibodies present (0, group I; 1, group II; 2, group III). Multiple ordinary least squares regression was used with the dependent variables to develop a model to predict bone mineral density (BMD) change.

Results

A total of 477 participants who completed a 3-year observation period were included. After matching, 312 participants were enrolled (group A, 104; group B, 208). Three years later, group B had significant BMD reduction in the femoral neck (FN) (p < 0.001), total hip (TH) (p = 0.001), and first through fourth lumbar vertebrae (L1–4) (p = 0.006), while group A had bone loss only at FN (p = 0.002). Groups I, II, and III included 104, 52, and 156 participants, respectively. Compared to baseline, BMD decreased significantly at FN (p = 0.002) in group I, FN (p < 0.001) in group II, and FN (p < 0.001), TH (p = 0.002), and L1–4 (p = 0.016) in group III. In terms of regression-adjusted percent change in BMD, more significantly negative changes were found at all measured sites in group B (p < 0.001, all) and at TH and L1–4 within groups I-III (p for trend < 0.001 and < 0.001, respectively). Regardless of antibodies, anti-osteoporotic therapy can preserve bone density in RA patients.

Conclusion

After 3 years, SPRA patients lost more bone density than SNRA patients. More attention should be paid to SPRA patients, especially those with double-positive antibodies, including a vigorous evaluation of BMD and fracture risk. Anti-osteoporotic therapy can prevent BMD loss irrespective of autoantibodies.
Tph Cells Expanded in Primary Sjögren’s Syndrome
Authors: Chen Weiqian , Yang Fan , Lin Jin
Published: June 2022

Objectives

PD-1+CXCR5–CD4+T peripheral helper cells, named Tph cells, contribute to B-cell immune responses and the production of antibodies in systemic lupus erythematosus and rheumatoid arthritis. However, the role of Tph cells was unknown in the pathogenesis of primary Sjögren’s syndrome (pSS). Here, we aim to explore the contribution of Tph cells in the development of pSS.

Methods

Sixty patients with pSS and 61 age and sex-matched healthy individuals were recruited for this study. The frequency of Tph cells in the blood was measured by flow cytometry. The expression of inducible T-cell costimulator (ICOS), MHC-II, IL-21, CCR2, CCR5, and CCR9 was evaluated in Tph cells. The relationship between Tph cells and indicators of clinical disease was assessed. Co-expression levels of PD-1, CXCR5, CD4, CCR2, and CCR5 in the salivary gland specimens from patients with pSS and patients with dry mouth and eyes but normal pathology were also analyzed.

Results

We demonstrated increased circulating Tph cells (7.53 ± 6.65% vs. 3.08 ± 1.31%, p < 0.0001) in patients with pSS (n = 60) compared to healthy controls (n = 61). Tph cells were significantly associated with the ESSDAI disease activity scores, IgG, ESR, IL-21, anti-SSA antibody, and CD138+/CD19+ plasma cells. Furthermore, ICOS was highly expressed in Tfh and Tph cells in patients with pSS. IL-21, MHC-II, CCR2, and CCR5 expression was higher in pSS Tph cells, and CCR9 expression was lower in pSS Tph cells than in pSS Tfh cells. Moreover, Tph cells and CCR2+CD4+T and CCR5+CD4+T cells were found in the labial gland of patients with pSS.

Conclusion

Our data show that Tph cells were enriched in peripheral blood and labial gland of patients with pSS. Circulating Tph cells correlated with disease activity scores, suggesting a crucial role of Tph in the development of pSS.
Back to the Roots of Nursing: Qualitative Study on the Experience of Nurses in the Front Line During the COVID-19 Pandemic
Authors: De Benedictis Anna , Gualandi Raffaella , Saccoccia Sabrina , Pensieri Claudio , Piredda Michela , De Micco Francesco , Marchetti Anna , Facchinetti Gabriella , Pasquarelli Alessia Assunta , De Carolis Chiara , Di Blasio Irene , Tartaglini Daniela , Alloni Rossana
Published: June 2022

The COVID-19 emergency has led many health facilities to reorganize themselves in a very short time to meet the urgent needs for intensive, semi-intensive or ordinary care of SARS-CoV-2 patients. In this pandemic, characterized by speed of transmission and severity of respiratory symptoms, care has been affected by the increase in volume and clinical complexity of patients, the sudden and unpredictable staff decrease and the lack of support from family members / caregivers. At the same time, experience in the field has shown how “informal” resources have been activated, which enabled to treat the highest possible number of patients above the real availability of resources. The purpose of this study was to explore the experiences of nurses involved in frontline care (COVID Centers) during the pandemic with a particular focus on professional motivation and on the development of technical-professional and personal skills. A study with a qualitative research design using focus group technique was conducted. Two focus groups were held with nine nurses. Data were analyzed with inductive content analysis. The findings can be summarized in five main categories: professional identity; motivation and sense of mission; development of professional and personal skills; spirituality; person-centered care; uniqueness of the lived experience. These findings shed new light on the correlation between motivation, professional identity and value, sense of duty and sense of belonging to the professional group. Moreover, the experience in the COVID Centers represented a valuable opportunity for participants to rediscover some specific issues related to nursing professional identity and to develop new personal and technical-professional skills in a very short time. Finally, nurses experienced once again how the nurse-patient relationship and basic care are essential to provide effective and excellent care, even and especially for patients in critical conditions. Nurses re-discovered, in a careful body care and basic care, irreplaceable elements to give back to patients, often dying, their own dignity, and all the needed closeness and attention necessary also to compensate the absence of the loved ones. These elements represent a way to concretely and deeply express the ethics of a job well done in nursing.
Impact of Probiotics on Dairy Production Efficiency
Authors: Nalla Kirankumar , Manda Naresh Kumar , Dhillon Harmeet Singh , Kanade Santosh R. , Rokana Namita , Hess Matthias , Puniya Anil Kumar
Published: June 2022

There has been growing interest on probiotics to enhance weight gain and disease resistance in young calves and to improve the milk yield in lactating animals by reducing the negative energy balance during the peak lactation period. While it has been well established that probiotics modulate the microbial community composition in the gastrointestinal tract, and a probiotic-mediated homeostasis in the rumen could improve feed conversation competence, volatile fatty acid production and nitrogen flow that enhances the milk composition as well as milk production, detailed changes on the molecular and metabolic level prompted by probiotic feed additives are still not understood. Moreover, as living biotherapeutic agents, probiotics have the potential to directly change the gene expression profile of animals by activating the signalling cascade in the host cells. Various direct and indirect components of probiotic approaches to improve the productivity of dairy animals are discussed in this review.
Evaluation and Genome Analysis of Bacillus subtilis YB-04 as a Potential Biocontrol Agent Against Fusarium Wilt and Growth Promotion Agent of Cucumber
Authors: Xu Wen , Yang Qian , Yang Fan , Xie Xia , Goodwin Paul H. , Deng Xiaoxu , Tian Baoming , Yang Lirong
Published: June 2022

Cucumber wilt caused by Fusarium oxysporum f.sp. cucumerinum (Foc) is a highly destructive disease that leads to reduced yield in cucumbers. In this study, strain YB-04 was isolated from wheat straw and identified as Bacillus subtilis. It displayed strong antagonistic activity against F. oxysporum f.sp. cucumerinum in dual culture and exhibited significant biocontrol of cucumber Fusarium wilt with a higher control effect than those of previously reported Bacillus strains and displayed pronounced growth promotion of cucumber seedlings. B. subtilis YB-04 could secrete extracellular protease, amylase, cellulose, and β-1,3-glucanase and be able to produce siderophores and indole acetic acid. Inoculation with B. subtilis YB-04 or Foc increased cucumber defense-related enzyme activities for PPO, SOD, CAT, PAL, and LOX. However, the greatest increase was with the combination of B. subtilis YB-04 and Foc. Sequencing the genome of B. subtilis YB-04 showed that it had genes for the biosynthesis of various secondary metabolites, carbohydrate-active enzymes, and assimilation of nitrogen, phosphorous, and potassium. B. subtilis YB-04 appears to be a promising biological control agent against the Fusarium wilt of cucumber and promotes cucumber growth by genomic, physiological, and phenotypic analysis.
Epidemiology and Clinical Features of Listeriosis in Gipuzkoa, Spain, 2010–2020
Authors: Vallejo Pedro , Cilla Gustavo , López-Olaizola Maddi , Vicente Diego , Marimón José María
Published: June 2022

Background

Listeriosis continues to be one of the most important notifiable foodborne diseases. Nonetheless, in Spain, there are few data on the molecular epidemiology of Listeria monocytogenes infections in recent years.

Aim

To describe clinical features and the molecular epidemiology of human listeriosis over an 11-year period (2010–2020) in Gipuzkoa, Northern Spain.

Methods

A total of 111 isolates, all but one from invasive disease, were studied. Serotyping (agglutination and multiplex polymerase chain reaction [PCR]) and multilocus sequence typing were performed for all isolates. Antibiotic susceptibility was assessed by the broth microdilution method.

Results

The average annual incidence of listeriosis in non-pregnancy-associated cases was 1.55 per 100,000 population, with a 1-month mortality rate of 22.2%. In pregnant women, the average incidence was 0.45 cases per 1,000 pregnancies. Twenty-four sequence types were identified, serotype 4b ST1 (24.3%) being the most frequent followed by 1/2b ST87 (18.9%), which caused two long outbreaks in 2013–2014. A significant association was observed between ST219 and meningitis (p < 0.001). All isolates were susceptible to ampicillin as well as other antibiotics used in listeriosis treatment.

Conclusion

Despite current control measures, listeriosis continues to be an important cause of mortality in the elderly, preterm birth, and miscarriages in pregnant women. Improvements in the control and diagnosis of listeriosis are needed to reduce the impact of this infection on vulnerable populations.
The Response of the Rodent Gut Microbiome to Broad-Spectrum Antibiotics Is Different in Males and Females
Authors: Parodi Gonzalo , Leite Gabriela , Pimentel Maya L. , Barlow Gillian M. , Fiorentino Alyson , Morales Walter , Pimentel Mark , Weitsman Stacy , Mathur Ruchi
Published: June 2022

Gut microbiome composition is different in males and females, but sex is rarely considered when prescribing antibiotics, and sex-based differences in gut microbiome recovery following antibiotic treatment are poorly understood. Here, we compared the effects of broad-spectrum antibiotics on both the stool and small bowel microbiomes in male and female rats. Adult male and female Sprague Dawley rats were exposed to a multi-drug antibiotic cocktail for 8 days, or remained unexposed as controls. Following cessation of antibiotics, rats were monitored for an additional 13-day recovery period prior to euthanasia. Baseline stool microbiome composition was similar in males and females. By antibiotic exposure day 8 (AbxD8), exposed male rats exhibited greater loss of stool microbial diversity compared to exposed females, and the relative abundance (RA) of numerous taxa were significantly different in exposed males vs. exposed females. Specifically, RA of phylum Proteobacteria and genera Lactobacillus, Sutterella, Akkermansia, and Serratia were higher in exposed males vs. exposed females, whereas RA of phyla Firmicutes and Actinobacteria and genera Turicibacter and Enterococcus were lower. By 13 days post antibiotics cessation (PAbxD13), the stool RA of these and other taxa remained significantly different from baseline, and also remained significantly different between exposed males and exposed females. RA of phyla Firmicutes and Actinobacteria and genus Enterococcus remained lower in exposed males vs. exposed females, and genus Sutterella remained higher. However, RA of phylum Proteobacteria and genus Akkermansia were now also lower in exposed males vs. females, whereas RA of phylum Bacteroidetes and genus Turicibacter were now higher in exposed males. Further, the small bowel microbiome of exposed rats on PAbxD13 was also significantly different from unexposed controls, with higher RA of Firmicutes, Turicibacter and Parabacteroides in exposed males vs. females, and lower RA of Bacteroidetes, Proteobacteria, Actinobacteria, Oscillospira, Sutterella, and Akkermansia in exposed males vs. females. These findings indicate that broad-spectrum antibiotics have significant and sex-specific effects on gut microbial populations in both stool and the small bowel, and that the recovery of gut microbial populations following exposure to broad-spectrum antibiotics also differs between sexes. These findings may have clinical implications for the way antibiotics are prescribed.
Functional Characterization of the Thrombospondin-Related Paralogous Proteins Rhoptry Discharge Factors 1 and 2 Unveils Phenotypic Plasticity in Toxoplasma gondii Rhoptry Exocytosis
Authors: Possenti Alessia , Di Cristina Manlio , Nicastro Chiara , Lunghi Matteo , Messina Valeria , Piro Federica , Tramontana Lorenzo , Cherchi Simona , Falchi Mario , Bertuccini Lucia , Spano Furio
Published: June 2022

To gain access to the intracellular cytoplasmic niche essential for their growth and replication, apicomplexan parasites such as Toxoplasma gondii rely on the timely secretion of two types of apical organelles named micronemes and rhoptries. Rhoptry proteins are key to host cell invasion and remodeling, however, the molecular mechanisms underlying the tight control of rhoptry discharge are poorly understood. Here, we report the identification and functional characterization of two novel T. gondii thrombospondin-related proteins implicated in rhoptry exocytosis. The two proteins, already annotated as MIC15 and MIC14, were renamed rhoptry discharge factor 1 (RDF1) and rhoptry discharge factor 2 (RDF2) and found to be exclusive of the Coccidia class of apicomplexan parasites. Furthermore, they were shown to have a paralogous relationship and share a C-terminal transmembrane domain followed by a short cytoplasmic tail. Immunofluorescence analysis of T. gondii tachyzoites revealed that RDF1 presents a diffuse punctate localization not reminiscent of any know subcellular compartment, whereas RDF2 was not detected. Using a conditional knockdown approach, we demonstrated that RDF1 loss caused a marked growth defect. The lack of the protein did not affect parasite gliding motility, host cell attachment, replication and egress, whereas invasion was dramatically reduced. Notably, while RDF1 depletion did not result in altered microneme exocytosis, rhoptry discharge was found to be heavily impaired. Interestingly, rhoptry secretion was reversed by spontaneous upregulation of the RDF2 gene in knockdown parasites grown under constant RDF1 repression. Collectively, our results identify RDF1 and RDF2 as additional key players in the pathway controlling rhoptry discharge. Furthermore, this study unveils a new example of compensatory mechanism contributing to phenotypic plasticity in T. gondii.
Removal of Dinotefuran, Thiacloprid, and Imidaclothiz Neonicotinoids in Water Using a Novel Pseudomonas monteilii FC02–Duckweed (Lemna aequinoctialis) Partnership
Authors: Cai Xiao-Yu , Xu Man , Zhu Yu-Xuan , Shi Ying , Wang Hong-Wei
Published: June 2022

Neonicotinoids (NEOs) are the most widely used insecticides in the world and pose a serious threat to aquatic ecosystems. The combined use of free-floating aquatic plants and associated microorganisms has a tremendous potential for remediating water contaminated by pesticides. The aim of this study was to determine whether plant growth-promoting bacteria (PGPB) could enhance the phytoremediation efficiency of duckweed (Lemna aequinoctialis) in NEO-contaminated water. A total of 18 different bacteria were isolated from pesticide-stressed agricultural soil. One of the isolates, Pseudomonas monteilii FC02, exhibited an excellent ability to promote duckweed growth and was selected for the NEO removal experiment. The influence of strain FC02 inoculation on the accumulation of three typical NEOs (dinotefuran, thiacloprid, and imidaclothiz) in plant tissues, the removal efficiency in water, and plant growth parameters were evaluated during the 14-day experimental period. The results showed that strain FC02 inoculation significantly (p < 0.05) increased plant biomass production and NEO accumulation in plant tissues. The maximum NEO removal efficiencies were observed in the inoculated duckweed treatment after 14 days, with 92.23, 87.75, and 96.42% for dinotefuran, thiacloprid, and imidaclothiz, respectively. This study offers a novel view on the bioremediation of NEOs in aquatic environments by a PGPB–duckweed partnership.
A Large Silicon Pool in Small Picophytoplankton
Authors: Wei Yuqiu , Sun Jun
Published: June 2022

Marine picophytoplankton (<2 μm) play a key role in supporting food web and energy flow in the ocean, and are major contributors to the global marine carbon (C) cycle. In recent years, picophytoplankton have been found to have significant silica (Si) accumulation, a finding which provides a new sight into the interaction of marine C and Si cycles and questions the overwhelming role of large diatoms (>2 μm) in the Si cycle. As picophytoplankton have high cell abundance and wide distribution in the open ocean, exploring their influences on the C and Si cycles as well as other element cycles are becoming new scientific hotspots. However, there are still few studies on the physiology and ecology of picophytoplankton, especially their potential roles in the biogeochemical Si cycle at present. Thus, it is necessary to accurately evaluate and quantify the contributions of picophytoplankton to the C and Si cycles, and to further understand their C and Si sinking mechanisms. In this review, we expect to have a novel understanding of picophytoplankton Si pool and regulation mechanism by conducting targeted studies on these scientific issues. This also provides a premise foundation and theoretical framework for further study of the role of small cells in the global ocean Si cycle and the coupling of C and Si cycles.
Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis
Authors: Qian Bingxu , Liao Kai , Zeng Dexin , Peng Wanqing , Wu Xiaodong , Li Jinming , Bo Zongyi , Hu Yongxin , Nan Wenlong , Wen Yuan , Cao Yuying , Xue Feng , Zhang Xiaorong , Dai Jianjun
Published: June 2022

Porcine epidemic diarrhoea virus (PEDV) is a member of the genus Alphacoronavirus in the family Coronaviridae. It causes acute watery diarrhoea and vomiting in piglets with high a mortality rate. Currently, the GII genotype, PEDV, possesses a high separation rate in wild strains and is usually reported in immunity failure cases, which indicates a need for a portable and sensitive detection method. Here, reverse transcription–recombinase aided amplification (RT-RAA) was combined with the Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas12a system to establish a multiplexable, rapid and portable detection platform for PEDV. The CRISPR RNA (crRNA) against Spike (S) gene of GII PEDV specifically were added into the protocol. This system is suitable for different experimental conditions, including ultra-sensitive fluorescence, visual, UV light, or flow strip detection. Moreover, it exhibits high sensitivity and specificity and can detect at least 100 copies of the target gene in each reaction. The CRISPR/Cas12a detection platform requires less time and represents a rapid, reliable and practical tool for the rapid diagnosis of GII genotype PEDV.
Efficiency of Antimicrobial Peptides Against Multidrug-Resistant Staphylococcal Pathogens
Authors: Le Mi Nguyen-Tra , Kawada-Matsuo Miki , Komatsuzawa Hitoshi
Published: June 2022

Antibiotics play a vital role in saving millions of lives from fatal infections; however, the inappropriate use of antibiotics has led to the emergence and propagation of drug resistance worldwide. Multidrug-resistant bacteria represent a significant challenge to treating infections due to the limitation of available antibiotics, necessitating the investigation of alternative treatments for combating these superbugs. Under such circumstances, antimicrobial peptides (AMPs), including human-derived AMPs and bacteria-derived AMPs (so-called bacteriocins), are considered potential therapeutic drugs owing to their high efficacy against infectious bacteria and the poor ability of these microorganisms to develop resistance to them. Several staphylococcal species including Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus saprophyticus are commensal bacteria and known to cause many opportunistic infectious diseases. Methicillin-resistant Staphylococci, especially methicillin-resistant S. aureus (MRSA), are of particular concern among the critical multidrug-resistant infectious Gram-positive pathogens. Within the past decade, studies have reported promising AMPs that are effective against MRSA and other methicillin-resistant Staphylococci. This review discusses the sources and mechanisms of AMPs against staphylococcal species, as well as their potential to become chemotherapies for clinical infections caused by multidrug-resistant staphylococci.
Aged Callus Skeletal Stem/Progenitor Cells Contain an Inflammatory Osteogenic Population With Increased IRF and NF-κB Pathways and Reduced Osteogenic Potential
Authors: Lin X. , Zhang H. , Liu J. , Wu C L. , McDavid A. , Boyce B. F. , Xing L.
Published: June 2022

Skeletal stem/progenitor cells (SSPCs) are critical for fracture repair by providing osteo-chondro precursors in the callus, which is impaired in aging. However, the molecular signatures of callus SSPCs during aging are not known. Herein, we performed single-cell RNA sequencing on 11,957 CD45-CD31-Ter119- SSPCs isolated from young and aged mouse calluses. Combining unsupervised clustering, putative makers, and DEGs/pathway analyses, major SSPC clusters were annotated as osteogenic, proliferating, and adipogenic populations. The proliferating cluster had a differentiating potential into osteogenic and adipogenic lineages by trajectory analysis. The osteoblastic/adipogenic/proliferating potential of individual clusters was further evidenced by elevated expression of genes related to osteoblasts, adipocytes, or proliferation. The osteogenic cluster was sub-clustered into house-keeping and inflammatory osteogenic populations that were decreased and increased in aged callus, respectively. The majority of master regulators for the inflammatory osteogenic population belong to IRF and NF-κB families, which was confirmed by immunostaining, RT-qPCR, and Western blot analysis. Furthermore, cells in the inflammatory osteogenic sub-cluster had reduced osteoblast differentiation capacity. In conclusion, we identified 3 major clusters in callus SSPCs, confirming their heterogeneity and, importantly, increased IRF/NF-κB-mediated inflammatory osteogenic population with decreased osteogenic potential in aged cells.
Characterization of Integrin Molecular Tension of Human Breast Cancer Cells on Anisotropic Nanopatterns
Authors: Kim Kyung Ah , Vellampatti Srivithya , Kim Byoung Choul
Published: June 2022

Physical interactions between cells and micro/nanometer-sized architecture presented in an extracellular matrix (ECM) environment significantly influence cell adhesion and morphology, often facilitating the incidence of diseases, such as cancer invasion and metastasis. Sensing and responding to the topographical cues are deeply associated with a physical interplay between integrins, ligands, and mechanical force transmission, ultimately determining diverse cell behavior. Thus, how the tension applied to the integrin-ligand bonds controls cells’ response to the topographical cues needs to be elucidated through quantitative analysis. Here, in this brief research report, we reported a novel platform, termed “topo-tension gauge tether (TGT),” to visualize single-molecule force applied to the integrin-ligand on the aligned anisotropic nanopatterns. Using the topo-TGT assay, first, topography-induced adhesion and morphology of cancerous and normal cells were compared with the pre-defined peak integrin tension. Next, spatial integrin tensions underneath cells were identified using reconstructed integrin tension maps. As a result, we characterized each cell’s capability to comply with nanotopographies and the magnitude of the spatial integrin tension. Altogether, the quantitative information on integrin tension will be a valuable basis for understanding the biophysical mechanisms underlying the force balance influencing adhesion to the topographical cues.
Current Advances in CETSA
Authors: Tolvanen Tuomas Aleksi
Published: June 2022

Knowing that the drug candidate binds to its intended target is a vital part of drug discovery. Thus, several labeled and label-free methods have been developed to study target engagement. In recent years, the cellular thermal shift assay (CETSA) with its variations has been widely adapted to drug discovery workflows. Western blot–based CETSA is used primarily to validate the target binding of a molecule to its target protein whereas CETSA based on bead chemistry detection methods (CETSA HT) has been used to screen molecular libraries to find novel molecules binding to a pre-determined target. Mass spectrometry–based CETSA also known as thermal proteome profiling (TPP) has emerged as a powerful tool for target deconvolution and finding novel binding partners for old and novel molecules. With this technology, it is possible to probe thermal shifts among over 7,000 proteins from one sample and to identify the wanted target binding but also binding to unwanted off-targets known to cause adverse effects. In addition, this proteome-wide method can provide information on the biological process initiated by the ligand binding. The continued development of mass spectrometry labeling reagents, such as isobaric tandem mass tag technology (TMT) continues to increase the throughput of CETSA MS, allowing its use for structure–activity relationship (SAR) studies with a limited number of molecules. In this review, we discussed the differences between different label-free methods to study target engagement, but our focus was on CETSA and recent advances in the CETSA method.
Legume Plant Peptides as Sources of Novel Antimicrobial Molecules Against Human Pathogens
Authors: Lima Rui M. , Rathod Balaji Baburao , Tiricz Hilda , Howan Dian H. O. , Al Bouni Mohamad Anas , Jenei Sándor , Tímár Edit , Endre Gabriella , Tóth Gábor K. , Kondorosi Éva
Published: June 2022

Antimicrobial peptides are prominent components of the plant immune system acting against a wide variety of pathogens. Legume plants from the inverted repeat lacking clade (IRLC) have evolved a unique gene family encoding nodule-specific cysteine-rich NCR peptides acting in the symbiotic cells of root nodules, where they convert their bacterial endosymbionts into non-cultivable, polyploid nitrogen-fixing cells. NCRs are usually 30–50 amino acids long peptides having a characteristic pattern of 4 or 6 cysteines and highly divergent amino acid composition. While the function of NCRs is largely unknown, antimicrobial activity has been demonstrated for a few cationic Medicago truncatula NCR peptides against bacterial and fungal pathogens. The advantages of these plant peptides are their broad antimicrobial spectrum, fast killing modes of actions, multiple bacterial targets, and low propensity to develop resistance to them and no or low cytotoxicity to human cells. In the IRLC legumes, the number of NCR genes varies from a few to several hundred and it is possible that altogether hundreds of thousands of different NCR peptides exist. Due to the need for new antimicrobial agents, we investigated the antimicrobial potential of 104 synthetic NCR peptides from M. truncatula, M. sativa, Pisum sativum, Galega orientalis and Cicer arietinum against eight human pathogens, including ESKAPE bacteria. 50 NCRs showed antimicrobial activity with differences in the antimicrobial spectrum and effectivity. The most active peptides eliminated bacteria at concentrations from 0.8 to 3.1 μM. High isoelectric point and positive net charge were important but not the only determinants of their antimicrobial activity. Testing the activity of shorter peptide derivatives against Acinetobacter baumannii and Candida albicans led to identification of regions responsible for the antimicrobial activity and provided insight into their potential modes of action. This work provides highly potent lead molecules without hemolytic activity on human blood cells for novel antimicrobial drugs to fight against pathogens.