Biology, Volume 10, Issue 11 (November 2021) – 149 articles

Endophytic fungi are proving to be an excellent source of chemical entities with unique structures and varied bioactivities. Terretonin (TE) and its structurally related derivatives are a class of meroterpenoids, possessing the same unique tetracyclic core skeleton, which have been reported from the Aspergillus genus. This study was carried out to assess the potential protective effects of TE separated from the endophytic fungus A. terreus against LPS (lipopolysaccharide)-induced ALI (acute lung injury) in mice. The results revealed that TE alleviated pulmonary edema as it lowered both the W/D lung ratio and protein content. The inflammatory response represented by inflammatory cell infiltration into the lung tissues was greatly repressed by TE. That was supported by the improved histopathological results and also by the reduced level of myeloperoxidase in the lung. TE showed a potent antioxidant activity as it attenuated lipid peroxidative markers (malondialdehyde, 4-hydroxynonenal, and protein carbonyl) and enhanced endogenous antioxidants (reduced glutathione, superoxide dismutase, and catalase) in lung tissues. Similarly, TE increased the mRNA expression of SIRT1, Nrf2, and its genes (HO-1, NQO1, and GCLm). On the other hand, TE restrained the activation of NF-κB (nuclear factor-κB) in the lung. Consequently, TE depressed the pro-inflammatory cytokines: nitric oxide (NOx), TNF-α (tumor necrosis factor-α), and interleukins (IL-6 and -1β). Additionally, TE inhibited NLRP3 signaling and interrupted apoptosis by decreasing the levels of proapoptotic markers (Bax and caspase-3) and increasing the level of an anti-apoptotic marker (Bcl-2). In conclusion, TE had a remarkable protective potential on LPS-induced lung damage via antioxidant and anti-inflammatory mechanisms. This finding encourages further investigations on this promising candidate. Full article

Although hemorrhagic sepsis caused by Aeromonas hydrophila infection is the dominant disease in the aquaculture of Chinese soft-shelled turtle, information on its molecular pathology is seriously limited. In this study, ninety turtles intraperitoneally injected with A. hydrophila exhibited two different phenotypes based on the pathological symptoms, referred to as active and inactive turtles. Comparative transcriptomes of liver and spleen from these two groups at 6, 24, and 72 h post-injection (hpi) were further analyzed. The results showed that cytokine–cytokine receptor interaction, PRRs mediated signaling pathway, apoptosis, and phagocytosis enriched in active and inactive turtles were significantly different. Pro-inflammatory cytokines, the TLR signaling pathway, NLR signaling pathway, and RLR signaling pathway mediating cytokine expression, and apoptosis-related genes, were significantly up-regulated in inactive turtles at the early stage (6 hpi). The significant up-regulation of phagocytosis-related genes occurred at 24 hpi in inactive turtles and relatively lagged behind those in active turtles. The anti-inflammatory cytokine, IL10, was significantly up-regulated during the tested periods (6, 24, and 72 hpi) in active turtles. These findings offer valuable information for the understanding of molecular immunopathogenesis after A. hydrophila infection, and facilitate further investigations on strategies against hemorrhagic sepsis in Chinese soft-shelled turtle T. sinensis. Full article

Human prostate cancer (PCa) is a type of malignancy and one of the most frequently diagnosed cancers in men. Elastin is an important component of the extracellular matrix and is involved in the structure and organization of prostate tissue. The present study examined prostate cancer in a xenograft mouse model using an elastin-specific molecular probe for magnetic resonance molecular imaging. Two different tumor sizes (500 mm³ and 1000 mm³) were compared and analyzed by MRI in vivo and histologically and analytically ex vivo. The T1-weighted sequence was used in a clinical 3-T scanner to calculate the relative contrast enhancement before and after probe administration. Our results show that the use of an elastin-specific probe enables better discrimination between tumors and surrounding healthy tissue. Furthermore, specific binding of the probe to elastin fibers was confirmed by histological examination and laser ablation–inductively coupled plasma–mass spectrometry (LA-ICP-MS). Smaller tumors showed significantly higher signal intensity (p > 0.001), which correlates with the higher proportion of elastin fibers in the histological evaluation than in larger tumors. A strong correlation was seen between relative enhancement (RE) and Elastica–van Gieson staining (R2 = 0.88). RE was related to inductively coupled plasma–mass spectrometry data for Gd and showed a correlation (R2 = 0.78). Thus, molecular MRI could become a novel quantitative tool for the early evaluation and detection of PCa. Full article

Ciliate infection is a serious parasitic disease of coral. Infected coral rots and dies in a short time. In addition to killing corals by infecting them in the oceans, ciliate infection also poses a threat to corals farmed on a large scale. In this study, two antioxidant enzymes (SOD and CAT) were used to judge the stress response in Goniopora columna after infection, and KCl and H₂O₂ were used to evaluate the therapeutic effect. The results showed that SOD and CAT increased during the early stage of infection but decreased with the extension of infection time. In terms of drug therapy, it was found that the treatment of ciliate infection with 1.5% of KCl had no significant effect on SOD and CAT of G. columna. The morphological changes of zooxanthellae, chlorophyll a, and coral were not significant. H₂O₂ leads to a stress response and polyp contraction. In conclusion, 1.5% of KCl can be used in the selection of drugs to treat ciliate infection. Full article

Intense physical activity contributes to an increased demand for red blood cells, which transport oxygen to working muscles. The purpose of this study was to assess the concentration of erythroferrone (ERFE), the novel marker of erythroid activity in athletes, during the beginning of their training season. The study group consisted of 39 athletes aged 23.24 ± 3.77 years. The study was carried out during the athletes’ preparatory period of the training cycle. The control group consisted of 34 healthy men aged 22.33 ± 2.77 years. The erythropoietic activity was evaluated by determining athletes’ concentrations of erythropoietin (EPO) and erythroferrone (ERFE). The level of physical activity was assessed using the International Physical Activity Questionnaire (IPAQ). In the athletes’ group, we observed higher concentrations of EPO (Me = 12.65 mIU/mL) and ERFE (40.00 pg/mL) compared to the control group (EPO: Me = 5.74 mIU/ml, p = 0.001; ERFE: Me = 25.50 pg/mL, p = 0.0034). The average intensity of physical exercise significantly differentiated the participants as far as EPO and ERFE concentrations. These results suggest that intense physical activity, at least at the beginning of the training season, may stimulate EPO production, which increases ERFE release. This seems to be an adaptative mechanism that provides adequate iron for enhanced erythropoiesis. Full article

The extracellular matrix (ECM) in granulosa cells is functionally very important, and it is involved in many processes related to ovarian follicle growth and ovulation. The aim of this study was to describe the expression profile of genes within granulosa cells that are associated with extracellular matrix formation, intercellular signaling, and cell–cell fusion. The material for this study was ovaries of sexually mature pigs obtained from a commercial slaughterhouse. Laboratory-derived granulosa cells (GCs) from ovarian follicles were cultured in a primary in vitro culture model. The extracted genetic material (0, 48, 96, and 144 h) were subjected to microarray expression analysis. Among 81 genes, 66 showed increased expression and only 15 showed decreased expression were assigned to 7 gene ontology groups “extracellular matrix binding”, “extracellular matrix structural constituent”, “binding, bridging”, “cadherin binding”, “cell adhesion molecule binding”, “collagen binding” and “cadherin binding involved in cell-cell adhesion”. The 10 genes with the highest expression (POSTN, ITGA2, FN1, LAMB1, ITGB3, CHI3L1, PCOLCE2, CAV1, DCN, COL14A1) and 10 of the most down-regulated (SPP1, IRS1, CNTLN, TMPO, PAICS, ANK2, ADAM23, ABI3BP, DNAJB1, IGF1) were selected for further analysis. The results were validated by RT-qPCR. The current results may serve as preliminary data for further analyses using in vitro granulosa cell cultures in assisted reproduction technologies, studies of pathological processes in the ovary as well as in the use of the stemness potential of GCs. Full article

The heterogeneous nature of ejaculates highlights the relevance of studying the behavior of different sperm subpopulations. Changes in sperm motility and the increase in tyrosine phosphorylation are key events that usually occur during capacitation and can be modified by the cryopreservation process. However, the relationship between both events remains poorly defined throughout capacitation in the different sperm subpopulations. Fresh and frozen-thawed spermatozoa were incubated in capacitating (CAP) and non-capacitating (NC) media up to 240 min. Sperm kinematics, tyrosine phosphorylation and mitochondrial activity were measured by the CASA system and imaging flow cytometry. Four motile sperm subpopulations (SP) were identified in fresh and frozen-thawed ram semen after the cluster analysis. Incubation under CAP conditions over time led to greater changes in the percentage of spermatozoa included in each subpopulation compared to NC conditions, being different between fresh and frozen-thawed spermatozoa. The SP1, characterized by slow spermatozoa, progressively increased after 15 min in frozen-thawed samples incubated in both media but not in fresh ones. The SP4, characterized by fast and non-linear spermatozoa, showed a marked increase during CAP, but not under NC conditions, occurring more rapidly in frozen-thawed spermatozoa. This subpopulation (SP4) was also the only one positively and strongly correlated with mitochondrial activity and all phosphorylated sperm regions during capacitation, either in fresh or frozen-thawed samples. Our results indicated that in vitro capacitation induced significant changes in the distribution of motile sperm subpopulations, being affected by cryopreservation. Notwithstanding, the subpopulation which probably represents hyperactivated-like spermatozoa (SP4) also increased in frozen-thawed samples, occurring faster and simultaneously to the increment of mitochondrial activity and tyrosine phosphorylation of different sperm regions. Full article

(1) Background: The control of angiogenesis is essential in disease treatment. We investigated angiogenesis-promoting or -suppressing factors and their molecular mechanisms. (2) Methods: Angiogenesis from HUVECs was quantitatively analyzed using the Angiogenesis Analysis Kit (Kurabo, Osaka, Japan). Human rAng-1-producing 107-35 CHO cells or mouse DFAT-D1 cells were co-cultured with HUVEC. Antioxidant polyphenols were added to the culture. Gene expression was analyzed by RT-PCR. (3) Results: The addition of rAng-1-producing cells, their culture supernatant, or commercially available rAng-1 showed a promoting effect on angiogenesis. The co-culture of DFAT-D1 cells promoted angiogenesis. Polyphenols showed a dose-dependent inhibitory effect on angiogenesis. Luteolin and quercetin showed remarkable anti-angiogenic effects. The expression of vWF, Flk1, and PECAM-1 was increased by adding rAng-1-producing cell culture supernatant. Polyphenols suppressed these genes. Apigenin and luteolin markedly suppressed α-SMA and Flk1. Resveratrol and quercetin enhanced the expression of PPARγ, and luteolin suppressed the expression of COX-1. The expression of endothelial nitric oxide synthase (eNOS), an oxidative stress-related gene, was slightly increased by luteolin. These results suggest that polyphenols induce ROS reduction. (4) Conclusions: We showed the promoting effect of Ang-1 or DFAT and the suppressing effect of polyphenols on angiogenesis and studied their molecular mechanisms. These results help control angiogenesis in regenerative therapy. Full article

Proteins from “pressure-loving” piezophiles appear to adapt by greater compressibility via larger total cavity volume. However, larger cavities in proteins have been associated with lower unfolding pressures. Here, dihydrofolate reductase (DHFR) from a moderate piezophile Moritella profunda (Mp) isolated at ~2.9 km in depth and from a hyperpiezophile Moritella yayanosii (My) isolated at ~11 km in depth were compared using molecular dynamics simulations. Although previous simulations indicate that MpDHFR is more compressible than a mesophile DHFR, here the average properties and a quasiharmonic analysis indicate that MpDHFR and MyDHFR have similar compressibilities. A cavity analysis also indicates that the three unique mutations in MyDHFR are near cavities, although the cavities are generally similar in size in both. However, while a cleft overlaps an internal cavity, thus forming a pathway from the surface to the interior in MpDHFR, the unique residue Tyr103 found in MyDHFR forms a hydrogen bond with Leu78, and the sidechain separates the cleft from the cavity. Thus, while Moritella DHFR may generally be well suited to high-pressure environments because of their greater compressibility, adaptation for greater depths may be to prevent water entry into the interior cavities. Full article

Background: Rhythmic gymnastics performance is characterized by technical elements involving flexibility, aerobic capacity and strength. Increased core strength in rhythmic gymnastics could lead to improved sporting performance. Objective: The aim of this study was to analyze the effect of 12 weeks of core muscle training on core muscle performance in rhythmic gymnasts. Methods: A randomized controlled study involving 24 rhythmic gymnastics was conducted. Participants were randomly assigned to a control group (CG; n = 12; age 13.50 ± 3.17 years) or a training group (TG; n = 12; age 14.41 ± 2.35 years). Body composition, isometric strength of trunk, core endurance and core muscle electromyographic activity were measured (EMG) after 12 weeks of core training. Independent sample t-tests were carried out to compare baseline values between groups. A two-way repeated-measures analysis of variance (ANOVA) (time × group) was applied. Results: The TG improved body composition, trunk lean mass (mean differences MD = −0.31; p = 0.040), lean mass (MD = 0.43; p = 0.037) and bone mass (MD = −0.06; p < 0.001) after training. Core training increased isometric strength of trunk, flexion test (MD = −21.53; p = 0.019) and extension test (MD = 22.7; p = 0.049), as well as the prone bridge core endurance test (MD = −11.27; p = 0.040). The EMG values also increased in the TG in prone bridge for front trunk (MD = −58.58; p = 0.026). Conclusions: Core strength training leads to improvements in body composition, as well as improvements in trunk strength and increases in muscle electromyographic activity. These improvements could therefore improve performance during competitive rhythmic gymnastics exercises. Full article

Background: Overweight and obesity are conditions associated with sedentary lifestyle and accumulation of abdominal fat, determining increased mortality, favoring chronic diseases, and increasing cardiovascular risk. Although the evaluation of body composition and fat distribution are highly relevant, the high cost of the gold standard techniques limits their wide utilization. Therefore, the aim of this work was to explore the relationships between simple anthropometric measures and BIA variables using multivariate linear regression models to estimate body composition and fat distribution in adults. Methods: In this cross-sectional study, sixty-eight adult individuals (20 males and 48 females) were subjected to bioelectrical impedance analysis (BIA), anthropometric measurements (waist circumference (WC), neck circumference (NC), mid-arm circumference (MAC)), allowing the calculation of conicity index (C-index), fat mass/fat-free mass (FM/FFM) ratios, body mass index (BMI) and body shape index (ABSI). Statistical analyzes were performed with the R program. Nonparametric Statistical tests were applied to compare the characteristics of participants of the groups (normal weight, overweight and obese). For qualitative variables, the Fisher’s exact test was applied, and for quantitative variables, the paired Wilcoxon signed-rank test. To evaluate the linear association between each pair of variables, the Pearson correlation coefficient was calculated, and Multivariate linear regression models were adjusted using the stepwise variable selection method, with Akaike Information Criterion (p ≤ 0.05). Results: BIA variables with the highest correlations with anthropometric measures were total body water (TBW), body fat percentage (BFP), FM, FFM and FM/FFM. The multiple linear regression analysis showed, in general, that the same variables can be estimated through simple anthropometric measures. Conclusions: The assessment of fat distribution in the body is desirable for the diagnosis and definition of obesity severity. However, the high cost of the instruments (dual energy X-ray absorptiometry, hydrostatic weighing, air displacement plethysmography, computed tomography, magnetic resonance) to assess it, favors the use of BMI in the clinical practice. Nevertheless, BMI does not represent a real fat distribution and body fat percentage. This highlights the relevance of the findings of the current study, since simple anthropometric variables can be used to estimate important BIA variables that are related to fat distribution and body composition. Full article

Smoking increases cardiovascular disease (CVD) risk by leading to endothelial injury. E-cigarettes remain a popular way to stop smoking. Evidence on their effect on cardiovascular health is growing but remains limited, particularly in the short-term. The main objective of this study was to compare short-term cardiovascular effects in smokers who quit smoking using e-cigarettes with or without nicotine or prescription nicotine replacement therapy (NRT). This was a single-centre (Sheffield, UK) pragmatic three-arm randomised controlled trial which recruited adult smokers (≥10 cigarettes per day), who were willing to attempt to stop smoking with support (n = 248). Participants were randomised to receive either: (a) behavioral support and e-cigarettes with 18 mg/mL nicotine (n = 84); (b) behavioral support and e-cigarettes without nicotine (n = 82); (c) behavioral support and NRT (n = 82). Flow Mediated Dilation (%FMD), peak cutaneous vascular conductance responses to acetylcholine (ACh) and sodium nitroprusside (SNP) and mean arterial pressure (MAP) were recorded at baseline and three days after stopping smoking. General Linear Models were used to compare changes between groups and changes from follow-up. Adjusting for baseline, at follow-up, all outcomes (for the 208 participants that completed the 3-day assessments) with the exception of SNP had improved significantly over baseline and there were no differences between groups (%FMD F = 1.03, p = 0.360, df = 2,207; ACh F = 0.172, p = 0.84, df = 2,207; SNP F = 0.382, p = 0.68, df = 2,207; MAP F = 0.176, p = 0.84, df = 2,207). For smokers ≥20 cigarettes per day, benefits were also pronounced. Smoking cessation showed positive cardiovascular impact even after a 3-day period and the effects did not differ between nicotine-containing e-cigarettes, nicotine-free e-cigarettes and NRT. Full article

FN-III proteins are widely distributed in mammals and are usually involved in cellular growth, differentiation, and adhesion. The FNDC5/irisin regulates energy metabolism and is present in different tissues (liver, brain, etc.). The present study aimed to investigate the physiochemical characteristics and the evolution of FN-III proteins and FNDC5/irisin as a ligand targeting the gonadal receptors including androgen (AR), DDB1 and CUL4 associated factor 6 (DCAF6), estrogen-related receptor β (ERR-β), estrogen-related receptor γ (ERR-γ), Krüppel-like factor 15 (KLF15), and nuclear receptor subfamily 3 group C member 1 (NR3C1). Moreover, the putative role of irisin in folliculogenesis and spermatogenesis was also elucidated. We presented the molecular structure and function of 29 FN-III genes widely distributed in the buffalo genome. Phylogenetic analysis, motif, and conserved domain pattern demonstrated the evolutionary well-conserved nature of FN-III proteins with a variety of stable to unstable, hydrophobic to hydrophilic, and thermostable to thermo-unstable properties. The comparative structural configuration of FNDC5 revealed amino acid variations but still the FNDC5 structure of humans, buffalo, and cattle was quite similar to each other. For the first time, we predicted the binding scores and interface residues of FNDC5/irisin as a ligand for six representative receptors having a functional role in energy homeostasis, and a significant involvement in folliculogenesis and spermatogenesis in buffalo. Full article

Background: Immune checkpoint inhibitors (ICI) have transformed the management of various cancers. Serious and potentially fatal cardiovascular toxicity, as well as a progression of atherosclerosis, have been described, mainly in elderly and comorbid patients. Methods: We investigated 117 arterial segments of 12 young (under 50 years of age), otherwise healthy lymphoma patients pre/post-ICI treatment using 2-[18F]fluorodeoxyglucose (FDG) positron emission tomography (PET). Maximum FDG standardized uptake values (SUV_max) and target-to-background ratios (TBRs) were calculated along arterial segments. Additionally, metabolic activities (SUV_max) of the bone marrow, spleen, and liver were analyzed. The levels of high-sensitivity C-reactive protein (hsCRP) were assessed. Results: ICI therapy induced arterial inflammatory activity, detected by increased TBR in arterial segments without pre-existing inflammation (TBRneg_pre = 1.20 ± 0.22 vs. TBRneg_post = 1.71 ± 0.45, p < 0.001), whereas already-inflamed lesions remained unchanged. Dormant calcified segments (Hounsfield Units-HU ≥ 130) showed a significant increase in TBR values after ICI treatment (TBRcalc_pre = 1.36 ± 0.38 vs. TBRcalc_post = 1.76 ± 0.42, p < 0.001). FDG uptake measured in other organs and hsCRP levels remained unchanged after ICI therapy. Conclusions: Although the effects of ICI therapy on arterial inflammation are still incompletely understood, cancer immunotherapy might be a critical moderator of atherosclerosis with a subsequently increased risk of future cerebro- and/or cardiovascular events in young oncological patients. Full article

Sleep disorders and depression are significant nonmotor symptoms (NMSs) of Parkinson disease (PD). However, few effective, evidence-proven medical treatments are available for alleviating these symptoms. Bright light therapy (BLT) is a well-established treatment for circadian rhythm sleep disorders and seasonal affective disorder. The present study conducted a literature review for the effect of BLT on PD, especially a meta-analysis of randomized controlled trials (RCTs). We searched for studies using the PubMed and Cochrane Library databases. The major outcomes were the effects on sleep and depression. The effect on motor symptoms was also analyzed as a secondary outcome. This study was registered with PROSPERO (CRD42020204454). Six studies were included in the literature review only, and the other five RCTs were included in the meta-analysis. Despite the positive effects of BLT on PD patients, which were demonstrated in noncontrolled studies, in the meta-analysis of the RCTs, BLT did not significantly improve the depressive symptoms (standardized mean difference (SMD): −0.15, 95% confidence interval (CI): −0.48 to 0.17, p = 0.36) and excessive daytime sleepiness (EDS) (SMD: −0.12, 95% CI: −0.49 to 0.25, p = 0.53) in PD patients. Regarding motor symptoms, no significant beneficial effects were conferred (SMD: −0.11, 95% CI: −0.44 to 0.21, p = 0.49). In conclusion, BLT did not significantly alleviate depression and sleepiness. The inconsistency between BLT protocols, such as the varied timing, dosages, and treatment durations, may render BLT’s efficacy difficult to demonstrate. The small effect size obtained from the present meta-analysis indicates that future RCTs are necessary, for which BLT protocols are standardized and more patients are enrolled to determine whether a significant therapeutic benefit was conferred. Full article

Mammalian circadian genes are capable of producing a self-sustained, autonomous oscillation whose period is around 24 h. One of the major characteristics of the circadian clock is temperature compensation. However, the mechanism underlying temperature compensation remains elusive. Previous studies indicate that a single clock gene may determine the temperature compensation in several model organisms. In order to understand the influence of each individual clock gene on the temperature compensation, twenty-three well-known mammalian clock genes plus Timeless and Myc genes were knocked out individually, using a powerful gene-editing tool, CRISPR/Cas9. First, Bmal1, Cry1, and Cry2 were knocked out as examples to verify that deleting genes by CRISPR is effective and precise. Cell lines targeting twenty-two genes were successfully edited in mouse fibroblast NIH3T3 cells, and off-target analysis indicated these genes were correctly knocked out. Through measuring the luciferase reporters, the circadian periods of each cell line were recorded under two different temperatures, 32.5 °C and 37 °C. The temperature compensation coefficient Q₁₀ was subsequently calculated for each cell line. Estimations of the Q₁₀ of these cell lines showed that none of the individual cell lines can adversely affect the temperature compensation. Cells with a longer period at lower temperature tend to have a shorter period at higher temperature, while cells with a shorter period at lower temperature tend to be longer at higher temperature. Thus, the temperature compensation is a fundamental property to keep cellular homeostasis. We further conclude that the temperature compensation is a complex gene regulation system instead of being regulated by any single gene. We also estimated the proliferation rates of these cell lines. After systematically comparing the proliferation rates and circadian periods, we found that the cell growth rate is not dependent on the circadian period. Full article

Irradiation of facial bones is associated with a lifelong risk of osteonecrosis. In a rat model, maxillae were exposed to a single 5 Gy dose of external beam radiation and orthodontic force was applied for 2 weeks on the first maxillary molar; control rats were treated identically without radiation. Tooth movement in irradiated jaws was 30% less than in controls, representing radiation-related damage. Micro-CT, histological, and molecular outcomes of orthodontic tooth movement were studied. Microstructurally, bone parameters (trabecular thickness, bone volume fraction, bone mineral density) were significantly affected by orthodontic force but not by radiation. Histological parameters were influenced only by orthodontic force, especially by an increase in osteoclasts. A molecular study revealed a differential distribution of cells expressing pre-osteoclast markers (RANK+—majority, CD11b+, CD14+—minority), with changes being influenced by orthodontic force (increased CD11b+ and CD14+ cells) and also by radiation (decreased RANK+ cells). The activation status of osteoclasts (TRAP staining) showed an orthodontic-force-related increase, which probably could not fully compensate for the radiation-associated impairment. The overall balance showed that orthodontic force had elicited a substantial microstructural, histological, and functional normalization process in irradiated maxillae but a radiation-induced impact was still conspicuous. Additional studies are needed to validate these findings. Full article

Bacterial biological control agents (BCAs) have been increasingly used against plant diseases. The traditional approach to manufacturing such commercial products was based on the selection of bacterial species able to produce secondary metabolites that inhibit mainly fungal growth in optimal media. Such species are required to be massively produced and sustain long-term self-storage. The endpoint of this pipeline is large-scale field tests in which BCAs are handled as any other pesticide. Despite recent knowledge of the importance of BCA-host-microbiome interactions to trigger plant defenses and allow colonization, holistic approaches to maximize their potential are still in their infancy. There is a gap in scientific knowledge between experiments in controlled conditions for optimal BCA and pathogen growth and the nutrient-limited field conditions in which they face niche microbiota competition. Moreover, BCAs are considered to be safe by competent authorities and the public, with no side effects to the environment; the OneHealth impact of their application is understudied. This review summarizes the state of the art in BCA research and how current knowledge and new biotechnological tools have impacted BCA development and application. Future challenges, such as their combinational use and ability to ameliorate plant stress are also discussed. Addressing such challenges would establish their long-term use as centerfold agricultural pesticides and plant growth promoters. Full article

Aims: Myocardial abnormalities are common during COVID-19 infection and recovery. We examined left (LV) and right (RV) ventricular longitudinal strain in patients who had recovered from COVID-19 and assessed the correlation with exercise capacity. Methods and results: One hundred and eighty-four consecutive patients with history of COVID-19 disease who had been referred to rest or stress echocardiography because of symptoms, mainly dyspnea and chest pain, were included in the study. These patients were compared to 106 patients with similar age, symptoms, and risk factor profile with no history of COVID-19 disease. Clinical and echocardiographic parameters, including strain imaging, were assessed. The patient’s age was 48 ± 12 years. Twenty-two patients had undergone severe disease. There were no differences in the LV ejection fraction and diastolic function between the groups. However, LV and RV global and free wall strain were significantly lower (in absolute numbers) in patients who had recovered form COVID-19 infection (−20.41 ± 2.32 vs −19.39 ± 3.36, p = 0.001, −23.69 ± 3.44 vs −22.09 ± 4.20, p = 0.001 and −27.24 ± 4.7 vs −25.43 ± 4.93, p = 0.021, respectively). Global Longitudinal Strain (GLS) < −20% was present in only 37% of post COVID-19 patients. Sixty-four patients performed exercise echocardiography. Patients with GLS < −20% had higher exercise capacity with higher peak metabolic equivalent and exercise time compared to patients with GLS ≥ −20% (12.6 ± 2 vs 10 ± 2.5 METss and 8:00 ± 2:08 vs 6:24 ± 2:03 min, p < 0.001 and p = 0.003, respectively). Conclusion: In patients, who had recovered from COVID-19 infection, both LV and RV strain are significantly lower compared to control patients. The exercise capacity of these patients correlates with LV strain values. Rest and stress echocardiography in patients with symptoms after COVID-19 infection may identify patients that need further follow up to avoid long term complications of the disease. These preliminary results warrant further research, to test the natural history of these findings and the need and timing of treatment. Full article

The progression of non-small cell lung cancer (NSCLC) is linked to epithelial-mesenchymal transition (EMT), a biologic process that enables tumor cells to acquire a migratory phenotype and resistance to chemo- and immunotherapies. Discovery of novel biomarkers in NSCLC progression is essential for improved prognosis and pharmacological interventions. In the current study, we performed an integrated bioinformatics analysis on gene expression datasets of TGF-β-induced EMT in NSCLC cells to identify novel gene biomarkers and elucidate their regulation in NSCLC progression. The gene expression datasets were extracted from the NCBI Gene Expression Omnibus repository, and differentially expressed genes (DEGs) between TGF-β-treated and untreated NSCLC cells were retrieved. A protein-protein interaction network was constructed and hub genes were identified. Functional and pathway enrichment analyses were conducted on module DEGs, and a correlation between the expression levels of module genes and survival of NSCLC patients was evaluated. Prediction of interactions of the biomarker genes with transcription factors and miRNAs was also carried out. We described four protein clusters in which DEGs were associated with ubiquitination (Module 1), regulation of cell death and cell adhesions (Module 2), oxidation-reduction reactions of aerobic respiration (Module 3) and mitochondrial translation (Module 4). From the module genes, we identified ten prognostic gene biomarkers in NSCLC. Low expression levels of KCTD6, KBTBD7, LMO7, SPSB2, RNF19A, FOXA2, DHTKD1, CDH1 and PDHB and high expression level of KLHL25 were associated with reduced overall survival of NSCLC patients. Most of these biomarker genes were involved in protein ubiquitination. The regulatory network of the gene biomarkers revealed their interaction with tumor suppressor miRNAs and transcription factors involved in the mechanisms of cancer progression. This ten-gene prognostic signature can be useful to improve risk prediction and therapeutic strategies in NSCLC. Our analysis also highlights the importance of deregulation of ubiquitination in EMT-associated NSCLC progression. Full article

The aims of this study are: (1) to identify morphological asymmetries in road cycling by using a novel 3D scanning method and electrical bioimpedance, (2) to investigate possible asymmetries in road cyclists of low (LPG) and high (HPG) performance group, (3) to compare the number of morphological asymmetries between HPG and LPG of cyclists, and (4) to explore correlations between asymmetry scores and competition performance. Body composition and 3D anthropometric measurements were conducted on 48 top-level male road cyclists (178.98 ± 5.39 cm; 68.37 ± 5.31 kg) divided into high (n = 22) and low (n = 26) performance groups. Competition performance (CP) is represented through racing points gathered at the end of the competition season. The latter was used to divide road cyclists into low- and high-performing groups. One-way ANOVA was used to determine differences between groups, while paired-samples T-test and Absolute Asymmetry index (AA) were calculated (p ≤ 0.05) for paired variables inside the groups, and the Spearman correlation coefficient was used to explore correlations between AA and CP. Results showed statistically significant differences between the left and right side of different body segments (16 paired variables) among low-performing road cyclists in five paired variables of the upper body: elbow girth (4.35, p = 0.000), forearm girth (6.31, p = 0.000), arm surface area (2.54, p = 0.018), and arm volume (2.71, p = 0.012); and six paired variables of the lower body: leg lean mass (5.85, p = 0.000), leg length (3.04, p = 0.005), knee girth (4.93, p = 0.000), calf girth (5.25, p = 0.000), leg surface area (4.03, p = 0.000), and leg volume (5.3, p = 0.000). Altogether, the high-performing group of road cyclists statistically differed only in 2 out of 16 paired variables of the upper body: elbow girth (4.93, p = 0.000) and in forearm girth (5.12, p = 0.000). Low- and high-performing groups were statistically significantly different in the asymmetry of leg lean mass F(1,46) = 6.25, p = 0.016 and asymmetry of the calf girth F(1,46) = 7.44, p = 0.009. AA of calf girth on the total sample (n = 48) showed a significant correlation with CP (r = −0.461; p = 0.001). In conclusion, the study’s main finding was that high-performance road cyclists are more symmetrical than the low-performance group, for which it is significant to have a higher amount of morphological asymmetries. Full article

This study investigated the dynamics of the accumulation and release of Hg²⁺ in lichens, using Evernia prunastri (L.) Ach. as a model species. Thalli were incubated with solutions containing 1, 10, and 100 µM Hg²⁺ and then exposed for 1, 2, 3, 6, 12, 18, and 24 months at the Botanical Garden of the University of Siena (a location free from local Hg sources). Lichen samples accumulated Hg proportionally to the exposure concentration, and after the exposure, reductions over time were evident, already starting from 1–2 months. After 24 months, samples released 72–74 (healthy thalli) to 94% (unhealthy thalli) of the accumulated Hg, but control values of untreated samples were never reached. Depending on the Hg content after the exposure, stable decreased concentrations were reached after 6–24 months. The results of this study highlight the ability of the lichen E. prunastri to reflect rapidly increasing environmental Hg concentrations, as well as to indicate an ameliorated situation (e.g., the closure of an Hg source). However, we have found evidence that an acute pollution episode can influence the content of Hg in lichens for several years. Full article

β₂-microglobulin (β2m), the light chain of the MHC-I complex, is associated with dialysis-related amyloidosis (DRA). Recently, a hereditary systemic amyloidosis was discovered, caused by a naturally occurring D76N β2m variant, which showed a structure remarkably similar to the wild-type (WT) protein, albeit with decreased thermodynamic stability and increased amyloidogenicity. Here, we investigated the role of the D76N mutation in the amyloid formation of β2m by point mutations affecting the Asp76-Lys41 ion-pair of WT β2m and the charge cluster on Asp38. Using a variety of biophysical techniques, we investigated the conformational stability and partial unfolding of the native state of the variants, as well as their amyloidogenic propensity and the stability of amyloid fibrils under various conditions. Furthermore, we studied the intermolecular interactions of WT and mutant proteins with various binding partners that might have in vivo relevance. We found that, relative to WT β2m, the exceptional amyloidogenicity of the pathogenic D76N β2m variant is realized by the deleterious synergy of diverse effects of destabilized native structure, higher sensitivity to negatively charged amphiphilic molecules (e.g., lipids) and polyphosphate, more effective fibril nucleation, higher conformational stability of fibrils, and elevated affinity for extracellular components, including extracellular matrix proteins. Full article

Endurance profile assessment is of major interest to evaluate the cyclist’s performance potential. In this regard, maximal oxygen uptake and functional threshold power are useful functional parameters to determine metabolic training zones (ventilatory threshold). The aim of this study was to evaluate and compare the physiological profile of different road cyclist age categories (Youth, Junior, and Under-23) to obtain the performance requirements. Sixty-one competitive road cyclists (15–22 years) performed a maximal incremental test on a bike in order to determine functional parameters (maximal fat oxidation zone, ventilatory thresholds, maximal oxygen uptake, and functional threshold power) and metabolic training zones. The results suggest major differences, with the Youth group showing clear changes in all metabolic zones except in fat oxidation. The main differences between Under-23 vs. Junior groups were observed in maximal relative power output (Under-23: 6.70 W·Kg⁻¹; Junior: 6.17 W·Kg⁻¹) and relative functional threshold power (Under-23: 4.91 W·Kg⁻¹; Junior: 4.48 W·Kg⁻¹). The Youth group physiological profile is clearly different to the other age categories. Some parameters normalized to body weight (maximal oxygen consumption, load and functional threshold power) could be interesting to predict a sporting career during the Junior and Under-23 stages. Full article

DNA topoisomerases are the enzymes that regulate DNA topology in all living cells. Since the discovery and purification of ω (omega), when the first were topoisomerase identified, the function of many topoisomerases has been examined. However, their ability to relax supercoiling and unlink the pre-catenanes of partially replicated molecules has received little attention. Here, we used two-dimensional agarose gel electrophoresis to test the function of three type II DNA topoisomerases in vitro: the prokaryotic DNA gyrase, topoisomerase IV and the human topoisomerase 2α. We examined the proficiency of these topoisomerases on a partially replicated bacterial plasmid: pBR-TerE@AatII, with an unidirectional replicating fork, stalled when approximately half of the plasmid had been replicated in vivo. DNA was isolated from two strains of Escherichia coli: DH5αF’ and parE10. These experiments allowed us to assess, for the first time, the efficiency of the topoisomerases examined to resolve supercoiling and pre-catenanes in partially replicated molecules and fully replicated catenanes formed in vivo. The results obtained revealed the preferential functions and also some redundancy in the abilities of these DNA topoisomerases in vitro. Full article

This study aimed to investigate the effects of three strains of probiotics, these being Lactobacillus rhamnosus GG, Bifidobacterium animalis subsp. Lactis BB-12 and Saccharomyces boulardii CNCM I-745, on the body weight, animal performances and blood parameters of rabbits (male and female) of the ITELV2006 strain. The supplementation of the feed of the rabbits with the three probiotic strains allowed observing positive effects on most of the biochemical and haematological parameters investigated during a period of 60 days (30 days of supplementation and 30 days without treatment). Further, there was a significant improvement in the body weight of the rabbits at the end of the experiment. The effect of the three probiotics investigated in this trial was found to be related to the sex of the rabbits and to the intake period (duration). Ultimately, these findings raise the possibility of using probiotics to investigate in an in-depth and specific manner based on fixed factors such as the strain, the gender and age of the animals, the main underlying mechanisms and effects, which would allow achieving optimal and adapted health benefits and sustainable production. In the context of animal production, it is worth investigating in a targeted study the effect of the three strains on muscle growth and development and finding evidence of the possible consequences on meat quality traits of the rabbits supplemented with probiotics. Full article

Many recent reports have demonstrated remarkable preservation of proteins in fossil bones dating back to the Permian. However, preservation mechanisms that foster the long-term stability of biomolecules and the taphonomic circumstances facilitating them remain largely unexplored. To address this, we examined the taphonomic and geochemical history of Tyrannosaurus rex specimen Museum of the Rockies (MOR) 1125, whose right femur and tibiae were previously shown to retain still-soft tissues and endogenous proteins. By combining taphonomic insights with trace element compositional data, we reconstruct the postmortem history of this famous specimen. Our data show that following prolonged, subaqueous decay in an estuarine channel, MOR 1125 was buried in a coarse sandstone wherein its bones fossilized while interacting with oxic and potentially brackish early-diagenetic groundwaters. Once its bones became stable fossils, they experienced minimal further chemical alteration. Comparisons with other recent studies reveal that oxidizing early-diagenetic microenvironments and diagenetic circumstances which restrict exposure to percolating pore fluids elevate biomolecular preservation potential by promoting molecular condensation reactions and hindering chemical alteration, respectively. Avoiding protracted interactions with late-diagenetic pore fluids is also likely crucial. Similar studies must be conducted on fossil bones preserved under diverse paleoenvironmental and diagenetic contexts to fully elucidate molecular preservation pathways. Full article

The genus Aeromonas has been recognised as an important pathogenic species in aquaculture that causes motile Aeromonas septicaemia (MAS) or less severe, chronic infections. This study compares the pathogenicity of the different Aeromonas spp. that were previously isolated from freshwater fish with signs of MAS. A total of 124 isolates of Aeromonas spp. were initially screened for the ability to grow on M9 agar with myo-inositol as a sole carbon source, which is a discriminatory phenotype for the hypervirulent A. hydrophila (vAh) pathotype. Subsequently, LD₅₀ of six selected Aeromonas spp_. were determined by intraperitoneal injection of bacterial suspension containing 10³, 10⁵, and 10⁷ CFU/mL of the respective Aeromonas sp. to red hybrid tilapias. The kidneys, livers and spleens of infected moribund fish were examined for histopathological changes. The screening revealed that only A. dhakensis 1P11S3 was able to grow using myo-inositol as a sole carbon source, and no vAh strains were identified. The LD_50–240h of A. dhakensis 1P11S3 was 10⁷ CFU/mL, while the non-myo-inositol utilizing A. dhakensis 4PS2 and A. hydrophila 8TK3 was lower at 10⁵ CFU/mL. Similarly, tilapia challenged with the myo-inositol A. dhakensis 1P11S3 showed significantly (p < 0.05) less severe signs, gross and histopathological lesions, and a lower mortality rate than the non-myo-inositol A. dhakensis 4PS2 and A. hydrophila 8TK3. These findings suggested that myo-inositol utilizing A. dhakensis 1P11S3 was not a hypervirulent Aeromonas sp. under current experimental disease challenge conditions, and that diverse Aeromonas spp. are of concern in aquaculture farmed freshwater fish. Therefore, future study is warranted on genomic level to further elucidate the influence of myo-inositol utilizing ability on the pathogenesis of Aeromonas spp., since this ability correlates with hypervirulence in A. hydrophila strains. Full article

Anthropogenic nitrogen (N) and sulfur (S) deposition can change above- and belowground biodiversity, including soil microbial diversity. The diversity of denitrifying microorganisms is of great significance to the calculation of the global nitrogen cycle and nitrogen flux. For a long time, nirK and nirS have been used as the functional genes to study denitrifying microorganisms, and have gradually become molecular markers for studying the composition and diversity of denitrifying bacteria. Here, three-time exposures to N and S applications (7, 30, and 60 days), were independently established. Additionally, the abundance, diversity, and structure of nirK- and nirS-type denitrifying communities were examined by sequencing analyses in response to three treatments, namely, N and S (T_N/S), sodium chloride (T_NaCl) and deionized water (pH = 7.0) (CK). Our results suggest that T_N/S led to higher electrical conductivity (EC), total nitrogen (TN), total organic carbon (TOC), nitrate nitrogen (NO₃⁻-N), ammonium nitrogen (NH₄⁺-N), and lower pH compared with T_NaCl and CK, which affected the diversity of nirK- and nirS-type denitrifying bacterial communities. We also observed that the nirK-type denitrifying community demonstrated a higher sensitivity to N and S additions. Overall, our results are important for the understanding of nitrogen in soil and N₂O emissions. Full article

The major priority of research in the present day is to conserve the environment by reducing GHG emissions. A proposed solution by an expert panel from 195 countries meeting at COP 21 was to increase global SOC stocks by 0.4% year⁻¹ to compensate for GHG emissions, the ‘4 per 1000′ agreement. In this context, the application of biocrusts is a promising framework with which to increase SOC and other soil functions in the soil–plant continuum. Despite the importance of biocrusts, their application to agriculture is limited due to: (1) competition with native microbiota, (2) difficulties in applying them on a large scale, (3) a lack of studies based on carbon (C) balance and suitable for model parameterization, and (4) a lack of studies evaluating the contribution of biocrust weathering to increase C sequestration. Considering these four challenges, we propose three perspectives for biocrust application: (1) natural microbiome engineering by a host plant, using biocrusts; (2) quantifying the contribution of biocrusts to C sequestration in soils; and (3) enhanced biocrust weathering to improve C sequestration. Thus, we focus this opinion article on new challenges by using the specialized microbiome of biocrusts to be applied in a new environment to counteract the negative effects of climate change. Full article