Objective: Otitis media (OM) is a common disease in childhood, but much less prevalent in adults, where it, however, tends to have a less benign course. The incidence of OM in children has decreased in recent years, which might be partly explained by the introduction of watchful waiting strategies, and by the introduction of conjugate pneumococcal vaccination (PCV) in children. Adult OM has been sparsely studied, and it is not known whether a herd immunity effect has been achieved after the introduction of PCV. Watchful waiting is usually not considered in adult OM patients. The purpose of this study was to use a national database to investigate the incidence of adult OM in Sweden during the last 20 years. Design: The yearly number of patients aged 20 and over, admitted to hospital with OM was obtained through a national database from 2000 to 2019. The corresponding numbers for outpatients were obtained between 2005 and 2019. Main outcome measures: incidence of acute otitis media in adults in in- and outpatient care. Results: Incidence rates decreased steadily during the study period. Comparing data on hospitalised patients from 2016-2019 to 2005-2008 (PCV was introduced in 2009), the incidence rate ratio was 0.49 (95% CI 0.45-0.53); p<0.0001. The corresponding figures for outpatients were 0.62 (95% CI 0.61-0.63); p<0.0001. The decrease in hospital admissions was most pronounced among younger patients, whereas the opposite was true for outpatient visits. Conclusions: There has been a continuous decrease in incidence of adult patients admitted to hospital with OM in Sweden during the last twenty years. Outpatient visits for OM have declined since 2009.

Root exudates can greatly modify microbial activity and soil organic matter (SOM) mineralization. However, the mechanism of root exudation and its stoichiometric ratio of C/N controlling upon paddy soil C mineralization are poorly understand. In this study, we used a mixture of glucose, oxalic acid, and alanine as root exudate mimics, employing three C/N stoichiometric ratios (CN6, CN10, and CN80) to explore the underlying mechanisms involved in C mineralization. The input of root exudates enhanced CO2 emission by 1.8–2.3-fold than that of the control. Artificial root exudates with low C/N ratios (CN6 and CN10) increased the metabolic quotient (qCO2) by 12% over those obtained at higher stoichiometric ratios (CN80 and C-only), suggesting a relatively high energy demand for microorganisms to acquire organic N from SOM by increasing N-hydrolase production. The stoichiometric ratios of enzymes (β-1,4-glucosidase to β-1,4-N-acetyl glucosaminidase) promoting organic C degradation compared to those involved in organic N degradation showed a significant positive correlation with qCO2; the stoichiometric ratios of microbial biomass (MBC/MBN) were positively correlated with carbon use efficiency. This suggests that root exudates with higher C/N ratios entail an undersupply of N for microorganisms, triggering the release of N-degrading extracellular enzymes. This in turn decreases SOM mineralization, implying the C/N ratio of root exudates to be a controlling factor. Our findings show that the C/N stoichiometry of root exudates controls C mineralization by the specific response of the microbial biomass through the release of C- and N-releasing extracellular enzymes to adjust for the microbial C/N ratio.

As in many other countries, Peru has the Water Quality Standard (WQS) as the primary tool for managing and diagnosing water resources. An analysis variable by variable to define water quality as poor or good was applied by setting concentration limits. A second group of tools commonly used are Biotic Indexes based on tolerance of benthic macroinvertebrates to pollution, that reflect the impacts caused by a group of variables, even though they cannot identify which variables determine the viability of the ecosystem. This research proposes to include the Stable States approach to evaluate the ecological integrity in central Andes rivers to explore an alternative approach with the capacity to represent a broader number of factors through multivariate analysis. A ten-year database of biological and physical-chemical variables measured in five Andean rivers were evaluated. Our results suggest these rivers fluctuate into two seasonal stable states (wet and dry season), accounting for approximately 31% of the system variability. In the wet season, the equilibrium of the state was dominated by the highest levels of suspended solids, turbidity, coliform, phosphorus, and some metals. During the dry season, the key variables were dissolved oxygen, flow, physical habitat, and biotic and diversity indexes. Likewise, there seems to be a third alternative state influenced by human pressures because of variables that exceed the WQS. Regarding water quality, the concentrations of coliforms, phosphorus, and lead usually exceeded the limits in two stations, but not every year. The ecological condition was better represented by ABI index than EPT.

ABSTRACT Objectives:To characterize the expression of ANGPTL4 in ovarian granulosa cells (GCs) and its association with polycystic ovary syndrome. Design: A retrospective study. Setting: University-based center for reproductive medicine. Participants:This study included 104 PCOS patients and 112 control women undergoing in vitro fertilization-embryo transfer (IVF-ET) from the reproductive hospital affiliated with Shandong University between 2019 and 2021. Methods: The mRNA expression of ANGPTL4 in GCs were assessed by reverse transcription and real-time quantitative (RT-q)PCR, then clinical information for these patients were reviewed and analyzed. Main outcome measures: ANGPTL4 expression in GCs in participants, correlation between ANGPTL4 expression level and metabolic characteristics of patients and predictive value of ANGPTL4 expression for PCOS. Results:The RT-qPCR results showed that ANGPTL4 expression in the control group is significantly lower than that in the PCOS group(P=0.000). It indicated positive association with AMH(r=0.211), HOMA-IR(r=0.174), LDL/HDL(r=0.176), ApoB/ApoAI(r=0.155) and TC/HDL(r=0.189). Additionally, the ANGPTL4 expression in the ovarian granulosa cells might be a independent factor in PCOS(OR:3.345, 95%CI:1.951–5.734) and served as a good predictor for PCOS (AUC0.704, 95%CI 0.633-0.774,P＜0.001). Conclusions:For the first time our study revealed on the higher ANGPTL4 expression in ovarian GCs with PCOS, and its association with glucose and lipid metabolism showed that ANGPTL4 might be a predictor for PCOS and play an important role in metabolism and pathogenesis of PCOS. Funding: National Key R&D Program of China (2018YFC1003202, 2016YFC1000604) and Taishan scholar project special funds (No. ts201712103). Key words: polycystic ovary syndrome, angiopoietin-like protein 4, mRNA, ovarian granulosa cell, glycolipid metabolism

Obstetric care for women that use antidepressants in pregnancyLine Kolding, MD, PhDVera Ehrenstein, MPH, DSc, ProfessorLars Pedersen, MSc, PhD, ProfessorPuk Sandager, MD, PhD, Associate ProfessorOlav B. Petersen, MD, PhD, ProfessorNiels Uldbjerg, MD, DMSc, ProfessorLars H. Pedersen, MD, PhD, ProfessorCorresponding:Lars Henning PedersenAarhus University Hospital / Aarhus UniversityPalle Juul-Jensens Blvd. 99, 8200 Aarhus N, DenmarkEmail: lhp@clin.au.dkPhone: +45 50526512We are grateful to Drs. Braillon and Bewley for their interest in our recent paper in the BJOG 1 and would like to elaborate on some of the important points they raise.We agree with Braillon and Bewley on the urgent need for improved pharmacovigilance of medication in pregnancy in general, and for antidepressants in particular. There are excellent international collaborations (e.g., the EuroCat) and local initiatives (e.g., the Swedish JanusInfo), but clinically we’re often forced to rely on very limited information indeed. Systematic international recording as suggested by Braillon and Bewley would represent an important step forward.On a smaller scale, we are establishing an automated surveillance system based on curated data that include information on both pre- and postnatally diagnosed malformations. We have, however, faced substantial legal and bureaucratic challenges, and have been forced to use data from the Central Denmark Region only, instead of national data. The surveillance system is consequently based on information on approx. 75,000 pregnancies, and even though it has the potential to aide clinical management, it is a drop in the ocean of the huge potential of for instance a comparable European collaboration.In our study, we used ≥2 redeemed prescriptions to define exposure with a prevalence 1.1%.1 The prevalence of pregnant women that redeemed ≥1 prescription was 3.2% (p. 3/ Table S1), and even though this is likely an overestimation due to non-adherence, the estimates are in line with previously reported prevalences in Scandinavia.2Braillon and Bewley emphasise the need to also consider non-pharmacological treatment of some pregnant women with depression and, further, to provide evidence-based and individualised treatment of women in the reproductive ages. Optimal individualised care will definitely result in non-pharmacological treatment of some pregnant women but, reversely, will cause yet other women to continue or initiate pharmacological treatment. This is in line with what is almost a truism in this field, that the potential harmful foetal effects must be balanced against the potential benefits of a pharmacological treatment, but it is no easy task. Pregnant women might overestimate the foetal risks associated with use of medication3 and discontinue important treatment, on the other hand some may use medication when there may be a better alternative for them. Regardless, we need to provide optimal obstetric care for the pregnant women that choose treatment with antidepressants. If our results are correct, prenatal follow-up of pregnant women treated with venlafaxine may include targeted foetal heart scans, even though the underlying causal explanation for the observed association with cardiac malformations is undetermined.1. Kolding L, Ehrenstein V, Pedersen L, Sandager P, Petersen OB, Uldbjerg N, et al. Antidepressant use in pregnancy and severe cardiac malformations: Danish register-based study. BJOG. 2021 May 25.2. Zoega H, Kieler H, Norgaard M, Furu K, Valdimarsdottir U, Brandt L, et al. Use of SSRI and SNRI Antidepressants during Pregnancy: A Population-Based Study from Denmark, Iceland, Norway and Sweden. PLoS One. 2015;10(12):e0144474.3. Wolgast E, Lindh-Åstrand L, Lilliecreutz C. Women’s perceptions of medication use during pregnancy and breastfeeding—A Swedish cross-sectional questionnaire study. Acta Obstetricia et Gynecologica Scandinavica. 2019;98(7):856-64.

CRISPR/Cas9 gene targeting technology has become the most widely used gene editing technology in both plants and animals. However, substantial off-target effect remains as a major imperfection hindering its further application. Here, Nicotiana benthamiana leaf cell-free system was used to simulate in vivo environment. And the effects of different CRISPR/Cas9 components on DNA stability in cell-free system were studied to explore possible mechanisms causing CRISPR off-target. The results showed that overexpressing Cas9, nCas9 and dCas9 significantly inhibited DNA cleavage in the cell extracts. While overexpressing RNPs accelerated the target DNA cleavage but inhibited non-target DNA digestion in cell extracts, overexpressing nRNP and dRNP blocked the cleavage of either target or non-target sequences. Meanwhile, analysis of whole-genome sequencing data from mice and rice edited by different CRISPR tools revealed that the main off-target mutations were SNVs (single nucleotide variants), rather than Indels (insertions and deletions) that were readily induced by DNA double-strand breaks. The off-target sites did not match the conventionally predicted places but were PAM-rich sites preferred. Our study suggests that PAM-dependent binding without cleavage of CRISPR/Cas9 to non-target sequences may increase off-target mutation risks through impeding the necessary cleavage process for repairing spontaneous or environmentally induced non-targeted DNA mutations.

Title Page Manuscript Title:Simultaneous atrial and ventricular pacing during narrow complex tachycardia – The Janus Response. Which one is true?

Background: Increasing interest in physiological pacing has been countered with challenges such as accurate lead deployment and increasing pacing thresholds with His-bundle pacing (HBP). More recently, left bundle branch area pacing (LBBAP) has emerged as an alternative approach to physiologic pacing. Objective: To compare procedural outcomes and pacing parameters at follow-up during initial adoption of HBP and LBBAP at a single center. Methods: Retrospective review, from September 2016 to January 2020, identified the first 50 patients each who underwent successful HBP or LBBAP. Pacing parameters were then assessed at first follow-up after implantation and after approximately one year, evaluating for acceptable pacing parameters defined as sensing R-wave amplitude >5 mV, threshold <2.5 V @ 0.5 ms and impedance between 400 and 1200 Ohms. Results: The HBP group was younger with lower ejection fraction compared to LBBP (73.2±15.3 vs 78.2±9.2 years, p=0.047; 51.0±15.9% vs 57.0±13.1%, p = 0.044). Post-procedural QRS widths were similarly narrow (119.8±21.2 vs. 116.7±15.2ms; p = 0.443) in both groups. Significantly fewer patients with HBP met the outcome for acceptable pacing parameters at initial follow-up (56.0% vs 96.4%, p = 0.001) and most recent follow-up (60.7% vs 94.9%, p = <0.001; at 399±259 vs. 228±124 days, p = <0.001). More HBP patients required lead revision due to early battery depletion (0 vs 13.3%, at an average of 664 days). Conclusion: During initial adoption, as compared with LBBAP, HBP is associated with a significantly higher frequency of unacceptable pacing parameters, energy consumption, and lead revisions.

Poultry and poultry-derived products such as meat and eggs are among the main sources of non-typhoidal Salmonella (NTS) transmission to the human. Therefore, we performed a systematic review and used random-effects meta-analyses to 1) estimate the prevalence of NTS in poultry samples from birds, products and subproducts, and environmental samples, 2) examine the diversity and frequency of their serovars, and 3) estimate the prevalence and profiles of antimicrobial resistance (AMR) in NTS isolates reported in studies from the Americas. We included 157 studies from 15 countries comprising 261,408 poultry samples and estimated an overall pooled prevalence of 17.9% (95% CI: 10.8–26.3) in birds, 21.8% (17.7–26.1) in products and subproducts, and 29.5% (24.2–35.1) in environmental samples. At the national level, the prevalence of NTS was heterogenous across countries with the highest values in Mexico, the USA, and Canada. In total, 131 serovars were identified from 13,388 isolates, Heidelberg, Kentucky, Enteritidis, and Typhimurium were the most prevalent in the overall top 10 ranking (range 6.5–20.8%). At the national level, Enteritidis and Typhimurium were identified in most of the countries, though with national differences in their ranks. The prevalence of AMR increased from 24.1% for 1 antibiotic, to 36.2% for 2-3 antibiotics, and 49.6% for ≥ 4 antibiotics. Kentucky, Heidelberg, Typhimurium, and Enteritidis were the serovars with the highest prevalence of AMR and tetracycline, ampicillin, streptomycin, ceftiofur, and amoxicillin-clavulanic acid were the top five antibiotics to which NTS isolates were resistant. In conclusion, NTS was distributed through the avian production chain with high and heterogenous values of prevalence in poultry samples. Besides, there were distinctive patterns of serovars distribution across countries and an alarming prevalence of AMR among zoonotic serovars.

We report the third case of lichen planus (LP) following COVID-19 BNT162b2 vaccination in a 59-year-old woman with previous LP. The reactivation of LP in patients with dormant LP, suggests possible vaccine-induced immune dysregulation. We suggest that the already described vaccine-induced up-regulation of Th1 response, may play a relevant role.

Thyrotoxicosis factitia is not very uncommon. There is a high likelihood of it being missed in our day-to-day clinical practice which can lead not only to significant patient morbidity but also put burden on health-care resources. We report a case masquerading as refractory thyrotoxicosis diagnosed to have uncontrolled hyperthyroidism.

Given the increased need for mechanical circulatory support and subsequent development of right ventricular assist devices (RVAD), appropriate imaging needs to be described to facilitate care in patients with cardiogenic shock and heart failure. We present three cases in which the upper esophageal aortic arch short axis (UE AA SAX) view on transesophageal echocardiography (TEE) was utilized to effectively image RVADs: to confirm normal positioning, to detect and guide repositioning, and to visualize malfunction. These cases support the importance of the UE AA SAX TEE view in RVAD outflow imaging and, when obtainable, should be included in routine RVAD assessment.

Globally-threatened freshwater mussels belonging to the order Unionida (Bivalvia) may be adversely affected by dense beds of submerged macrophytes that modify habitat at the sediment-water interface. Such effects can be particularly pronounced in modified lentic ecosystems such as reservoirs which are subject to hydrological regimes (e.g., hydropeaking) that can exacerbate macrophyte-mediated impacts, including anoxic or hypoxic conditions, the related release of toxic ions (e.g., ammonia), and silt accumulation that inhibits filter-feeding. Accordingly, we compared how population size-structure and biomass of the New Zealand mussel Echyridella menziesii varied inside and outside of dense beds of invasive macrophytes known to have similar impacts on water chemistry (e.g., anoxia) in two northern New Zealand hydroreservoir locations with contrasting hydrology (lacustrine location dominated by Ceratophyllum demersum; and riverine location dominated by Egeria densa). We found adverse sediment-water interface conditions were not always associated with dense submerged macrophyte beds in littoral zones. Nonetheless, where they occurred, adverse sediment-water interface conditions were primary drivers in reduced mussel density and adult skewed size-structure, inferring reduced recruitment. Disentangling direct and indirect effects with structural equation modelling indicated that increased pore-water ammonia did not impact these primarily adult populations of freshwater mussels. Increased sediment organic matter, silt, and previously recorded hypoxia and anoxia were exacerbated in the lacustrine section where variable flows promoting water mixing were not present to reduce their effects. High densities of mussels less than 40 mm in length were associated with favourable sediment-water interface conditions of low silt and sediment organic matter, suggesting that enhanced water exchange in and around macrophyte beds may increase juvenile mussel survival in littoral zones of the riverine lake section. Our findings highlight a potential role for hydropeaking management in mitigating the development of adverse physicochemical conditions, and underscore the context-specific effects that dense non-native macrophyte beds can have on mussel populations.

Objectives To explore the impact of COVID-19 on the management and outcomes of acute paediatric mastoiditis across the UK. Design National retrospective and prospective audit Setting 48 UK secondary care ENT departments Participants Consecutive children aged 18 years or under, referred to ENT with a clinical diagnosis of mastoiditis. Main outcome measures Cases were divided into: Period 1 (01/11/19-15/03/20) before the UK population were instructed to reduce social contact, and Period 2 (16/03/20-30/04/21), following this. Periods 1&2 were compared for population variables, management and outcomes. Secondary analyses compared outcomes by primary treatment (medical/needle aspiration/surgical). Results 286 cases met criteria (median 4 per site, range 0-24). 9.4 cases were recorded per week in period 1 versus 2.0 in period 2, with no winter increase in cases in Dec 2020-Feb 2021. Patient age differed between period 1&2 (3.2 Vs 4.7 years respectively, p<0.001). 85% of children in period 2 were tested for COVID-19 with a single positive test. In period 2 cases associated with P. aeruginosa significantly increased. 48.6% of children were scanned in period 1 vs 41.1% in period 2. Surgical management was used more frequently in period 1 (43.0% Vs 24.3%, p=0.001). Treatment success was high, with failure of initial management in 6.3%, and 30-day re-admission for recurrence in 2.1%. The adverse event rate (15.7% overall) did not vary by treatment modality or between periods 1& 2. Conclusion The COVID-19 pandemic led to a significant change in the presentation and case-mix of acute paediatric mastoiditis in the UK.

Organogels are semisolid and 3D systems in which both an organic liquid solvent and other dispersed particles are entrapped. Today there is not much information about the potential of organogels as carriers of lipophilic nutraceuticals. Therefore, in this work, the preparation and characterization of soybean oil-based organogels with candelilla wax (CW) and lecithin, tocopherol, or EPA/DHA were studied. Soybean oil was structured with 0.5, 1.0, 2.0, and 4.0% w/w of CW. In the organogels, the visual appearance, thermal stability, color parameters, microstructure, hardness, and rheological properties were analyzed. Results showed that soybean oil was successfully structured into an array of solid-like organogels varying in concentrations of CW and added nutraceutical compounds. Increasing CW concentration yielded stronger 3D-network gel formation (type 5 or solid organogel). All organogels were stable at 5, 25, and 35°C during the cyclization process. The nutraceuticals affected the color parameters and 3D network structure. Larger crystalline spherulites and a uniform microstructure were observed, especially in organogels formulated with 4.0% w/w of CW and lecithin. In comparison with the control, organogels (4.0% w/w) with tocopherol or EPA/DHA required lower shear stress to flow whereas lecithin required higher shear stress to flow. Texture analysis showed that organogels with nutraceuticals presented higher hardness, this behavior was attributed to the formation of a larger and softer crystal networks. In conclusion, soybean oil-based organogels formulated with lecithin, tocopherol, or EPA/DHA yielded stable organogels with promising properties as nutraceutical carriers.

Since the first in-human implantation, trans-catheter aortic valve replacement (TAVR) has shown an exciting development in both technical and technological terms, becoming the standard of care for many patients, even not only inoperable ones. Although trans-femoral (TF) access has the scepter of first-line route for TAVR, in some cases, this access is not feasible, so several alternative routes were introduces over time. The network meta-analysis by Hameed et al has the great merit to provide a comprehensive picture. Hence, through either direct and indirect comparison, the authors confirmed as TF is the gold standard as access, followed by trans-carotid and trans-subclavian. Conversely, trans-thoracic (trans apical and trans-aortic) routes are the least safe and should be reserved only to sporadic cases.

The adsorption and fixation of potassium in agricultural soils are important as they influence K availability for crops. Soil organic matter (SOM) and ammonium (NH4+) exist in soils and play indispensable roles in soil fertility and crop yield; however, the effects of SOM and NH4+ on K retention in soil aggregate remains unclear. This study aimed to evaluate the effects of SOM and NH4+ on K adsorption and fixation in soil microaggregates (<0.25 mm). Soil microaggregates were extracted from three long-term fertilization treatments under rice-wheat rotations: no fertilizer (CK), fertilized with inorganic NPK (NPK), and inorganic NPK fertilizers combined with straw return (NPKS). Long-term fertilization, particularly the application of inorganic NPK combined with straw return, significantly improved the SOM content in microaggregates. Both NPK and NPKS treatments increased K adsorption but decreased K fixation, and SOM oxidation of microaggregates reduced K adsorption but increased K fixation in all treatments, indicating the positive and inhibitory effects of SOM on K adsorption and fixation, respectively. NH4+ significantly inhibited K adsorption and fixation, and this inhibitory effect was more significant in microaggregates with a higher SOM content. Although NH4+ reduced the positive effect of SOM on K adsorption, it enhanced the inhibitory effect of SOM on K fixation. Conclusionally, long-term fertilization increases K adsorption but reduces K fixation by improving SOM content, where NH4+ enhances SOM inhibited K retention in soil microaggregates, which is considered to improve K availability in soils amended with K fertilizers. Keywords: soil organic matter, NH4+, K, adsorption, fixation

We study the small initial date Cauchy problem for the generalized incompressible Navier-Stokes-Coriolis equations in critical hybrid-Besov space $\dot{\mathscr{B}}_{2,\, p}^{\frac{5}{2}-2\alpha, \frac{3}{p}-2\alpha+1}(\mathbb{R}^3)$ with $1/2<\alpha<2$ and $2\leq p\leq 4$. We prove that hybrid-Besov spaces norm of a class of highly osillating initial velocity can be arbitrarily small. and we prove the estimation of highly frequency $L^p$ smoothing effect for generalized Stokes-Coriolis semigroup with $1\leq p\leq\infty$, At the same time, we prove space-time norm estimation of hybrid-Besov spaces for Stokes-Coriolis semigroup. From this result we deduce bilinear estimation in our work space. Our method relies upon Bony’s high and low frequency decomposition technology and Banach fixed point theorem.

Coherent mapping with CARTO3 is useful to identify the critical isthmus of scar-related AT. However, it has also a pitfall. We present atrial tachycardia (AT) with pseudo micro-reentrant activation pattern created by coherent mapping on the right atrial free wall. It is possible that a pseudo tachycardia circuit was created due to the algorithm for the reconstruction of coherent mapping. Finally, entrainment mapping led to the identification of correct tachycardia circuit and termination of tachycardia by catheter ablation. When using coherent mapping with CARTO3, it is necessary to understand the pitfall well.

The complexity and uncertainty of land use and environmental factors pose challenges to the management decisions of ecological restoration and conservation.We integrated the mixed-cell CA model and Bayesian belief networks to develop an innovative method for optimizing ecosystem services under different land development scenarios, including consideration of the uncertainty and variability of factors.The southern region of Sichuan Province, China, was selected as an example.We first established three development scenarios between 2015 and 2035, namely, natural development scenario (NDS), ecological protection scenario (EPS), and cultivated land protection scenario (CLPS).The MCCA model was utilized to simulate the land use pattern in 2035 under different scenarios.We then construced a BBN-based model to investigate the carbon sequestration, grain supply, soil conservation, habitat quality, and water yield in 2035 under uncertain scenarios.After the sensitivity analysis and evaluation of the model, we determined the state combination of influential factors at various ecosystem service levels and the ecological restoration and conservation key areas.The obtained result showed that the key influencing factors impacting the ecosystem services level included NPP, Slope, forestland and ET, and the state combination corresponding to the highest level of ecosystem services was predominantly distributed in regions with the highest NPP, the highest Slope, the highest forestland area and low ET.Based on this finding, we proposed some suggestions for ecological restoration and conservation of key areas.This model considers uncertainties and is capable of providing scientific recommendations on restoration and conservation; therefore, it can serve as an effective tool to assist stakeholders in making decisions.

The concentration and homeostasis of intracellular phosphate (Pi) are crucial for sustaining cell metabolism and growth. During short-term Pi starvation, intracellular Pi is maintained relatively constant at the expense of vacuolar Pi. After the vacuolar stored Pi is exhausted, the plant cells induce the synthesis of intracellular acid phosphatase (APase) to recycle Pi from expendable organic phosphate (Po). In this study, the expression, enzymatic activity and subcellular localization of ACID PHOSPHATASE 1 (OsACP1) were determined. OsACP1 expression is specifically induced in almost all cell types of leaves and roots under Pi stress conditions. OsACP1 encodes an acid phosphatase with broad Po substrates and localizes in the endoplasmic reticulum (ER) and Golgi apparatus (GA). Phylogenic analysis demonstrates that OsACP1 has a similar structure with human acid phosphatase PHOSPHO1. Overexpression or mutation of OsACP1 affected Po degradation and utilization, which further influenced plant growth and productivity under both Pi-sufficient and Pi-deficient conditions. Moreover, overexpression of OsACP1 significantly affected intracellular Pi homeostasis and Pi starvation signalling. We concluded that OsACP1 is an active acid phosphatase that regulates rice growth under Pi stress conditions by recycling Pi from Po in the ER and GA.

Aims. The management of patients with asymptomatic significant aortic regurgitation (sAR) is often challenging and appropriate timing of aortic valve surgery remains controversial. Prognostic value of diastolic parameters has been demonstrated in several cardiac diseases. In particular, left atrial (LA) function has been shown to be an important determinant of morbimortality. The purpose of this study was to analyze the prognostic significance of diastolic function in asymptomatic patients with sAR. Methods and results. A total of 126 patients with asymptomatic sAR were included. Conventional echocardiographic systolic and diastolic function parameters were assessed. LA auto-strain analysis was performed in a subgroup of 57 patients. During a mean follow up of 33±19 months, 25 (19,8%) patients reached the combined end-point. Univariate analysis showed that LV volumes, LVEF, E wave, E/e’ ratio, LA volume and LA reservoir strain (LASr) were significant predictors of events. Multivariate model 1 that tested all echocardiographic variables statistically significant in the univariate model showed that LVEDV [HR=1,02;95% CI:1,01-1,03 (p<0,001)] and E/e’ ratio [HR=1,12;95% CI:1,03-1,23 (p=0,01)] were significant predictors of events. In the subgroup of patients with LA auto-strain analyzed, a second multivariate model including the previous significant variables for the first model as well as LASr, showed that LVEDV [HR=1,05;95% CI:1,01-1,08 (p=0,016)] and LASr [HR=0,8;95% CI:0,65-0,98 (p<0,035)] were the most significant predictors of cardiovascular events. Conclusions. In this population of asymptomatic patients with sAR and normal LV systolic function, baseline diastolic parameters were prognostic markers of cardiovascular events; among them, LASr played a strong independent predictor role.

Background and Purpose Growing evidence demonstrates that the delta opioid receptor (DOP) is an attractive candidate for novel antidepressants with the potential to exhibit rapid action with few adverse effects. However, the underlying detailed functional mechanism remains elusive. Previously, we reported that the selective DOP agonist, KNT-127, produced robust antidepressant-like effects in the mice forced swimming test (FST). Thus, we attempted to identify the cellular mechanism underlying this effect. Experimental Approach Male ICR mice (4–6 weeks) were used in all experiments. The FST was conducted as a screening model for antidepressants. The phosphorylation level of proteins in specific brain regions was quantified using Western blotting. Glutamate/gamma-aminobutyric acid-dependent postsynaptic currents were detected using whole-cell voltage-clamp recordings. Key Results The selective mTOR inhibitor, rapamycin, and the PI3K inhibitor, LY294002, blocked the antidepressant-like effects of KNT-127 in the FST. KNT-127 increased the phosphorylation level of mTOR signal-related proteins, Akt and p70S6K, in the medial prefrontal cortex. The bilateral microinfusion of KNT-127 in the infralimbic cortex decreased immobility in the FST. The frequency of miniature excitatory postsynaptic currents in the infralimbic cortex increased and that of miniature inhibitory postsynaptic currents decreased with the perfusion of KNT-127, which was blocked by pretreatment with rapamycin. Conclusions and Implications KNT-127 displays antidepressant-like actions through the direct facilitation of neuronal excitability in the mice infralimbic cortex, which is implicated in the PI3K-Akt-mTOR-p70S6K signaling pathway. These results could indicate the first steps in elucidating the complete mechanical functions of DOPs as a potential candidate for novel antidepressants.

G protein-coupled receptors modulate a plethora of physiological processes and mediate the effects of one-third of FDA-approved drugs. Notably, depending on which ligand has activated a particular receptor, it can engage different intracellular transducers. This paradigm of ligand-dependent ‘biased signaling’ dictates a need to advance beyond the level of receptors to consider the combined ligand-receptor pair in order to understand physiological signaling. Bias signaling also has the potential to improve medicines by reducing adverse effects. However, this is challenged by inconsistent interpretation of results and lack of commonly agreed guidelines. Here, we present recommended terminology and guidelines to conduct, report and quantify bias in a comparable and reproducible fashion. We expect these recommendations will facilitate a common understanding of experiments and findings across basic receptor research and drug discovery, while the area and the analytical methods to measure bias are still evolving, especially in complex cellular, tissue and organismal systems.