Acidic rain has been identified as potentially harmful to the aquatic and terrestrial components of the ecosystem. Sulphate measured in rain and snow has been used as a surrogate indicator of acidic deposition. If sulphur dioxide controls are the means to limit acidic deposition, then the association between sulphate and hydrogen ion concentrations in precipitation is an important factor in establishing such limits. Selected data on rain and snowfall chemistry from the National Atmospheric Deposition Program (NADP), the Electric Power Research Institute's SURE, the utility industries' UAPSP, and the Department of Energy's MAP3S were reviewed. Numerical analyses were performed to assess the relationship between hydrogen and sulphate ion concentrations. The strength of the association between hydrogen and sulphate ions varied from site to site. In the Midwestern and Eastern regions, the Pearson correlation coefficient was over 0.50 while in the Central and Upper Midwestern parts of the United States, the correlation coefficients were less than 0.25. Regardless of the strength of the association between hydrogen and sulphate ions, all but one of the NADP/NTN sites used in our analysis exhibited at least 30% of the anions (sulphate, nitrate, and chloride) associated with cations other than hydrogen. For sites where the strength of the association was weak, between 65% and 98% of the anions appeared to be associated with cations other than hydrogen. Because a large percentage of the anions (i.e. sulphate, nitrate, and chloride) appear to be associated with cations other than hydrogen even at those sites where the association between hydrogen and sulphate ions was strong, the complex chemistry controlling the acidity in precipitation may make it difficult to predict the impact of a reduction in sulphate concentration.

N-acylethanolamine acid amidase (NAAA) is an N-terminal nucleophile (Ntn) hydrolase that catalyses the intracellular deactivation of the endogenous analgesic and anti-inflammatory agent palmitoylethanolamide (PEA). NAAA inhibitors counteract this process and exert marked therapeutic effects in animal models of pain, inflammation and neurodegeneration. While it is known that NAAA preferentially hydrolyses saturated fatty acid ethanolamides (FAEs), a detailed profile of the relationship between catalytic efficiency and fatty acid-chain length is still lacking. In this report, we combined enzymatic and molecular modelling approaches to determine the effects of acyl chain and polar head modifications on substrate recognition and hydrolysis by NAAA. The results show that, in both saturated and monounsaturated FAEs, the catalytic efficiency is strictly dependent upon fatty acyl chain length, whereas there is a wider tolerance for modifications of the polar heads. This relationship reflects the relative stability of enzyme-substrate complexes in molecular dynamics simulations.

Keywords: N-acylethanolamine acid amidase; enzyme kinetic; molecular dynamics; palmitoylethanolamide; substrate selectivity.

Non-terrestrial networks (NTN) are expected to play a key role in extending and complementing terrestrial 5G networks in order to provide services to air, sea, and un-served or under-served areas. This paper focuses the attention on the uplink, where terminals are able to establish a direct link with the NTN at Ka-band. To reduce the collision probability when a large population of terminals is transmitting simultaneously, we propose a grant-free access scheme called resource sharing beamforming access (RSBA). We study RBSA for low Earth orbit (LEO) satellite communications with massive multiple-input multiple-output (MIMO). The idea is to benefit from the spatial diversity to decode multiple overlapped signals. We have devised a blind and open-loop beamforming technique, where neither the receiver must carry out brute-force search in azimuth and elevation, nor are the terminals required to report channel state information. Upon deriving the theoretical throughput, we show that RBSA is appropriate for grant-free access to LEO satellite, it reduces the probability of collision, and thus it increases the number of terminals that can access the media. Practical implementation aspects have been tackled, such as the estimation of the required statistics, and the determination of the number of users.

Keywords: LEO; beamforming; grant-free; massive IoT; massive MIMO; non-orthogonal multiple access; orthogonal frequency division multiplexing.

Objectives: Cardiothoracic training during the COVID-19 pandemic witnessed a dramatic change in the UK, resulting in changes in surgical provisions, centralisation of services and reduced surgical case volume. The aim of this study is to assess the impact COVID-19 on surgical training and seek opinions regarding the future role of surgical simulation in cardiothoracic training.

Methods: A cross sectional survey was designed and distributed amongst cardiothoracic national and non-national trainees in the UK. The survey consisted of 31 questions, split into four sections; Demographics, Service Provision, Training and Simulation Training.

Results: A total of 83/200 (41.5%) trainees completed the survey comprised of 44/129 (34.1%) national trainees (NTN) and 39/71 (54.9%) non-national trainees (non-NTN). 80.7% (67/83) of trainees agreed that cardiothoracic training had been negatively impacted as a result of COVID-19, p<0.001. Additionally, 12% agreed that adequate resources were available to learning/practise of technical skills. 87% agreed that simulation had a role in Cardiothoracic training, 81% recognised simulations is an important tool in improving their surgical skills and 79.5% agreed that simulation should be used to meet the increasing need in training/education moving forward.

Conclusions: COVID-19 has had a significant impact on surgical training, with concerns that these effects could have further implications downstream. Simulation training has been under-used thus far and trainees face an uphill challenge to enhance their skills and technical abilities in the operating room. Simulation is recommended by trainees and may represent one of the solutions to the challenges of safe and effective cardiothoracic surgical training.

Keywords: CCT, Certificate of Completion of Training; CESR, Certificate of Eligibility for Specialist Registration; COVID-19; COVID-19, Coronavirus disease 2019; Cardiothoracic Surgery; GMC, General Medical Council; JCST, Joint Committee on Surgical Training; NTN, national trainees (National Training Number); Non-NTN, non-national trainees; SCTS, Society for Cardiothoracic Surgery in Great Britain and Ireland; ST, Speciality Trainee; Simulation Training; Surgical Education; Surgical Training; Survey.

Type 2 asparaginases, a subfamily of N-terminal nucleophile (Ntn) hydrolases, are activated by limited proteolysis. This activation yields a heterodimer and a loop region at the C-terminus of the α-subunit is released. Since this region is unresolved in all type 2 asparaginase crystal structures but is close to the active site residues, we explored this loop region in six members of the type 2 asparaginase family using homology modeling. As the loop model for the childhood cancer-relevant protease Taspase1 differed from the other members, Taspase1 activation as well as the conformation and dynamics of the 56 amino acids loop were investigated by CD and NMR spectroscopy. We propose a helix-turn-helix motif, which can be exploited as novel anticancer target to inhibit Taspase1 proteolytic activity.

To elucidate the role of reactive oxygen species (ROS) in accelerated nephrotoxic nephritis (NTN), effects of SOD-mimic (Fe-TPAA) and anti-PAF (WEB-2086) on NTN were investigated. Three days after the preimmunization with normal rabbit IgG, anti-GBM rabbit IgG were administered unilaterally to the left kidney in order to examine the effect of circulating inflammatory factors by comparing with the contralateral kidney. Normal rabbit IgG were likewise injected to the control rats. Luminol amplified chemiluminescence (CL) assay of isolated glomeruli, histological examination and monoclonal antibody (anti LC-A Ab, anti PMN Ab, anti M phi Ab) positive-cell counting were performed in both kidneys. After induction of NTN, 3.4 mg/kg/day of Fe-TPAA and 2mg/kg/day of WEB-2086 were respectively administered by continuous injection using Osmotic pump. At day 7, glomerular CL and glomerular cellularity were both increased significantly in anti-GBM perfused kidney. Anti-rabbit IgG were also stained in the contralateral kidney, but clearly in lesser amount as compared with the anti-GBM perfused kidney, suggesting very slight glomerulonephritis may have occurred also in nonperfused kidney. While WEB-administration suppressed glomerular hypercellularity, CL and urinary protein, Fe-TPAA administration did only glomerular CL. Taking into account that dosing amount of Fe-TPAA was limited due to its toxicity, the effective ROS scavenging may not have been obtained. However, another possibility is proposed that WEB-2086 suppressed the development of glomerular lesion by not only inhibiting generation of ROS but also by release of other inflammatory factors.

Single aphids can simultaneously or sequentially acquire and transmit multiple potato virus Y (PVY) strains. Multiple PVY strains are often found in the same field and occasionally within the same plant, but little is known about how PVY strains interact in plants or in aphid stylets. Immuno-staining and confocal microscopy were used to examine the spatial and temporal dynamics of PVY strain mixtures (PVY^O and PVY^NTN or PVY^O and PVY^N) in epidermal leaf cells of 'Samsun NN' tobacco and 'Goldrush' potato. Virus binding and localization was also examined in aphid stylets following acquisition. Both strains systemically infected tobacco and co-localized in cells of all leaves examined; however, the relative amounts of each virus changed over time. Early in the tobacco infection, when mosaic symptoms were observed, PVY^O dominated the infection although PVY^NTN was detected in some cells. As the infection progressed and vein necrosis developed, PVY^NTN was prevalent. Co-localization of PVY^O and PVY^N was also observed in epidermal cells of potato leaves with most cells infected with both viruses. Furthermore, two strains could be detected binding to the distal end of aphid stylets following virus acquisition from a plant infected with a strain mixture. These data are in contrast with the traditional belief of spatial separation of two closely related potyviruses and suggest apparent non-antagonistic interaction between PVY strains that could help explain the multitude of emerging recombinant PVY strains discovered in potato in recent years.

Keywords: Myzus persicae stylet; PVY; confocal microscopy; potato; recombinant strain; strain interaction.

In vitro diagnostics (IVD) has huge potential. Primary drivers in the global market are the patient's awareness of infectious diseases, the introduction of advanced molecular and tissue diagnostic tests for patient-stratified and targeted anti-cancer therapy and, last but not least, the growing geriatric population. Rapid progress in device miniaturization and information technology (IT) offers new possibilities in decentralized testing. Grand View Research Inc. expects the global market for IVD to reach US $ 74.3 billion by 2020. Hence the launch in 2015 by the NTN Swiss Biotech - together with the driving forces of Biotechnet Switzerland - of the 'Thematic Platform in vitro Diagnostics'.

The monitoring of hypertension (HTN) in dialysis is often delicate with potentially false measurements due to the white coat effect on the one hand and masked HTN (M-HTN) on the other hand. In this population, there is much controversy over the ideal moment for taking blood pressure (BP) and the target values. An answer to these questions is given by home BP measurement that can detect white coat HTN (WC-HTN) and M-HTN. The aim of this study was to determine the respective prevalence of permanent HTN (P-HTN), WC-HTN, M-HTN, and permanently normotensive (P-NTN) in this population and to analyze the risk factors of M-HTN and WC-HTN in hemodialysis (HD) centers in sub-Saharan Africa. This was a multicenter, descriptive, and analytical cross-sectional study conducted over a period of one month and 23 days. Data collection was performed using a home BP measurement form, conventional BP measurement form, and clinical and laboratory data collection form. The study included all patients who could take their BP at home using an electronic BP machine and record results on the BP forms. All analyses were performed using the Sphinx plus software version 5. The significance level for all statistical tests was set at 5%. The mean age of patients was 45.57 years ± 14.11, with a sex ratio of 1.42. The mean duration in dialysis was 57.96 months ± 34.86. Adherence to the home BP measurement was 100% in 71.7%. P-NTN patients were 15.2% (7 patients), WC-HTN patients were 13% (6 patients), M-HTN patients were 17.5% (8 patients), and P-HTN patients were 54.3% (25 patients). A statistically significant association was observed between WC-HTN and age (P = 0.01). In this work, we noted an important proportion of M-HTN and WC-HTN. This result confirms the need for home BP measurement in the follow-up of BP in HD patients.

Understanding the interactive effects of different warming levels and tillage managements on crop morphological and physiological traits and radiation use efficiency (RUE) is essential for breeding climate-resilient cultivars. Here, we conducted temperature free-air controlled enhancement (T-FACE) experiments on winter wheat during two growth seasons in the North China Plain. The experiments consisted of three warming treatments and two tillage treatments (CT: conventional tillage & NT: no-tillage). In the normal season, warming had significant positive effects on major morphological and physiological traits and increased significantly RUE of yield (RUE_Y ) and biomass (RUE_DM ) by 13.3% and 11.3%, 19.3% and 12.4%, 42.3% and 43.7%, respectively, under the treatments of CTT1, CTT2, and NTT1 relative to the control (CTN, NTN). By contrast, in the warmer season, warming had negative effects on leaf width, light extinction coefficient, light-saturated net photosynthetic rate, aboveground, stems and spike biomass, and RUE from anthesis to maturity, and consequently grain yield under conventional tillage, but positive effects under no-tillage. Our findings bring new insights into the mechanisms on the interactive effects of warming and tillage treatments on wheat growth and productivity; provide valuable information on crop ideotypic traits for breeding climate-resilient crop cultivars. This article is protected by copyright. All rights reserved.

Keywords: Climate-resilient cultivars; Growth; Heat; T-FACE; Triticum aestivum L; leaf photosynthesis.

Caspase-3 is the crucial executor caspase of apoptosis in mammalian cells, which is essential for chromatin condensation and DNA fragmentation. Although plants have no caspase-3 homologs, PBA1 acts as a plant caspase-3-like enzyme in plant programmed cell death (PCD). PCD occurs during the formation of secretory cavities in Citrus fruits; hence, secretory cavities could be utilized as a new cell biology model for investigating the regulatory mechanisms of plant PCD. To further study the association between PBA1 and PCD during secretory cavity development in Citrus fruits, CgPBA1 was identified in the fruit of Citrus grandis 'Tomentosa'. The temporal and spatial expression of CgPBA1 during secretory cavity development were analyzed using quantitative real-time PCR and in situ hybridization, and the morphological changes in the apoptotic cell nuclei were observed using TUNEL assay and ultra-thin section technology. The results revealed that the full-length cDNA of CgPBA1 contains a 711 bp ORF that encodes a putative protein containing 236 amino acid with a proteasome-β-6 functional domain that belongs to the Ntn hydrolase super family. CgPBA1 was predominantly expressed in the secretory cavities; its expression changes coincided with the morphological changes and DNA fragmentation in apoptotic cell nuclei. The green fluorescent fusion protein of CgPBA1 is also located in the nucleus of tobacco epidermal cells. Based on previous research and the findings of the present study, we speculate that CgPBA1 is a highly functional conserved protein in plants, and it might be involved in nuclear degradation during PCD for secretory cavity formation in C. grandis 'Tomentosa' fruits.

Keywords: CgPBA1; Nucleus degradation; PCD; Secretory cavity.

Basal Cell Carcinoma (BCC) is one of the most diagnosed cancers worldwide. It develops due to an unrestrained Sonic Hedgehog (SHH) signaling activity in basal cells of the skin. Certain subtypes of BCC are more aggressive than others, although the molecular basis of this phenomenon remains unknown. We have previously reported that Neogenin-1 (NEO1) is a downstream target gene of the SHH/GLI pathway in neural tissue. Given that SHH participates in epidermal homeostasis, here we analyzed the epidermal expression of NEO1 in order to identify whether it plays a role in adult epidermis or BCC. We describe the mRNA and protein expression profile of NEO1 and its ligands (Netrin-1 and RGMA) in human and mouse control epidermis and in a broad range of human BCCs. We identify in human BCC a significant positive correlation in the levels of NEO1 receptor, NTN-1 and RGMA ligands with respect to GLI1, the main target gene of the canonical SHH pathway. Moreover, we show via cyclopamine inhibition of the SHH/GLI pathway of ex vivo cultures that NEO1 likely functions as a downstream target of SHH/GLI signaling in the skin. We also show how Neo1 expression decreases throughout BCC progression in the K14-Cre:Ptch1lox/lox mouse model and that aggressive subtypes of human BCC exhibit lower levels of NEO1 than non-aggressive BCC samples. Taken together, these data suggest that NEO1 is a SHH/GLI target in epidermis. We propose that NEO1 may be important in tumor onset and is then down-regulated in advanced BCC or aggressive subtypes.

Background: Children with beta-thalassemia major (β-TM) suffer from tubular dysfunction even before the onset of any renal impairment symptoms and/or clinical signs. Therefore, identifying innovative biomarkers allowing early renal damage detection has focused attention.

Aim: This study aims to preliminary assess Netrin-1(NTN-1) and Clusterin (CLU) in β-TM children and explore their possible roles as surrogate noninvasive biomarkers of renal tubular dysfunction.

Subjects and methods: In this study, 40 β-TM children and 30 healthy children were enrolled. Routine serum and urinary biochemical variables were determined. Urinary NTN-1 and CLU levels were measured using ELISA and their mRNA expression in PBMCs were assayed using real-time PCR. Serum TNF-α, MDA levels and GST activity were measured.

Results: Urinary NTN-1 and CLU concentrations and mRNA relative expression levels in PBMCs were significantly increased in β-TM children relative to controls. Oxidative stress and inflammatory markers revealed significant elevation in β-TM children compared to controls. The change in these parameters correlated significantly with other renal parameters. ROC curves analysis showed that urinary NTN-1 and CLU levels are of promising diagnostic performance.

Conclusion: Our results suggest that NTN-1 and CLU are qualified as new noninvasive biomarker panels for early detection of renal injury in β-TM children. Moreover, urinary NTN-1 is recommended as a precise one during the clinical practices.

Keywords: Beta Thalassemia Major; Clusterin; GST; Netrin-1; Renal dysfunction; TNF-α.

Following nephron reduction by means of five--sixths nephrectomy (Nx), nephritis was induced in rats by injection of nephrotoxic serum (NTS). A continuous intravenous infusion of synthetic rat--atrial natriuretic peptide (r-ANP) at 2.5 micrograms/min for produced remarkable diuresis and natriuresis in these rats. Furthermore, inulin clearance, para-amino-hippuric acid clearance and filtration fraction increased in spite of the reduction of mean arterial pressure during the infusion of r-ANP. It was therefore evident that diuresis and natriuresis were mediated by a direct action of r-ANP in the kidney. We observed that r-ANP was a diuretic, natriuretic and hypotensive agent but at a higher dosage in Nx rats with NTN.

Background: It is currently not known how many trainees leave vascular surgery, and their reasons for doing so are unclear. This paper is the first to publish the number of UK trainees leaving the training programme and interrogates their reasons for doing so.

Methods: An email survey was distributed to current and recent Training Programme Directors (TPDs) to quantify the number of trainees resigning between 2013 and 2019. Trainees resigning a National Training Number (NTN) were surveyed regarding their reasons for doing so.

Results: Since 2013, 23 UK vascular surgery trainees have resigned NTNs, representing 15.4% of the 149 NTNs awarded between 2013 and our analysis. Reasons for leaving, as relayed by TPDs, included availability of an academic career, geography, health and many other reasons classified as "work-life balance" factors. Data from the trainees surveyed also highlighted work-life balance but also identified pressures within the training system and NHS.

Conclusions: UK data of this sort has not previously been available. The authors' primary recommendation is that prospective data collection on trainee retention is carried out, with structured exit interviews with trainees who decide to leave. Our secondary recommendations include improvements to the inter-deanery transfer process and early realistic exposure to vascular surgery for junior doctors to improve trainee retention rates in vascular surgery.

Keywords: Surgical training; Trainee retention; Vascular surgery.

We performed several experiments using three strains of Virgibacillus salexigens, namely, P2, NT N53, and C-20Mo^T (DSM 11483^T), which were isolated from completely different sources, in relation to bacteriocin production ability. Results of whole-genome sequencing analysis revealed that all strains have very similar sequences encoding class IId bacteriocin. Although a partial amino acid sequence of the purified bacteriocin produced by strain P2 isolated from fermented food was previously reported, whole-genome sequencing and the N-terminal sequencing results in this study showed that its complete amino acid sequence consisted of 48 residues, which corresponded to that of the hypothetical bacteriocin encoded by the gene in Virgibacillus massiliensis strain Vm-5^T (DSM 28587^T) isolated from the human gut. From the results of 16S rRNA gene sequencing and whole-genome sequencing analyses, we taxonomically confirmed Vm-5^T to be a strain of V. salexigens, and its broth culture showed antibacterial activity. Strain NT N53 isolated from the deep-sea floor produced two bacteriocins, namely, NTN-A and NTN-B. The results of N-terminal sequencing, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and whole-genome sequencing analyses showed that their amino acid sequences differed in only one residue, and NTN-A showed the same sequence as the bacteriocin produced by strain P2. Although strain C-20Mo^T isolated from a solar saltern had the coding sequence very similar to that of NTN-A, its broth culture showed no antibacterial activity. This finding suggests that class IId bacteriocin-producing or bacteriocin-gene-encoding V. salexigens strains are widely distributed in distinct environment sources with different geographical and material properties.

Keywords: Antibacterial substance; Bacteriocin; Different isolation sources; Virgibacillus salexigens.

In recent years, several recombinant strains of potato virus Y, notably PVY^NTN and PVY^N:O have displaced the ordinary strain, PVY^O, and emerged as the predominant strains affecting the USA potato crop. Previously we reported that recombinant strains were transmitted more efficiently than PVY^O when they were acquired sequentially, regardless of acquisition order. In another recent study, we showed that PVY^NTN binds preferentially to the aphid stylet over PVY^O when aphids feed on a mixture of PVY^O and PVY^NTN. To understand the mechanism of this transmission bias as well as preferential virus binding, we separated virus and active helper component proteins (HC), mixed them in homologous and heterologous combinations, and then fed them to aphids using Parafilm sachets. Mixtures of PVY^O HC with either PVY^N:O or PVY^NTN resulted in efficient transmission. PVY^N:O HC also facilitated the transmission of PVY^O and PVY^NTN, albeit with reduced efficiency. PVY^NTN HC failed to facilitate transmission of either PVY^O or PVY^N:O. When PVY^O HC or PVY^N:O HC was mixed with equal amounts of the two viruses, both viruses in all combinations were transmitted at high efficiencies. In contrast, no transmission occurred when combinations of viruses were mixed with PVY^NTN HC. Further study evaluated transmission using serial dilutions of purified virus mixed with HCs. While PVY^NTN HC only facilitated the transmission of the homologous virus, the HCs of PVY^O and PVY^N:O facilitated the transmission of all strains tested. This phenomenon has likely contributed to the increase in the recombinant strains affecting the USA potato crop.

Keywords: Myzus persicae; PVY; aphid; helper component; interaction; potato; strain shift; virus concentration.

Forest management practices play an important role in regulating the soil water-holding capacity of plantation. However, most studies focus on soil water dynamics present during large-scale forest loss and afforestation events, while little is known about how soil water under different forest management practices responds to rainfall events and which factors mainly regulate soil water-holding capacity. In this study, a stable hydrogen isotope was used to explore the contribution of three natural rainfall events (8.9, 13.3 and 67.7 mm) to soil water (CRSW) in a Pinus massoniana plantation under four forest management practices (no thinning (NTN), understory removal (USR), light-intensity thinning (LIT) and heavy-intensity thinning (HIT)) in the Three Gorges Reservoir Area of the Yangtze River Basin in China. Furthermore, a structural equation model was employed to determine the effects of vegetation biomass and soil properties on the CRSW. The results showed that plantation soil under different forest management practices exhibited different water-holding capacities. Following light (8.9 mm) and moderate (13.3 mm) rainfall events, the CRSW in the HIT stand was slightly higher than that in the other stands. Following heavy (66.7 mm) rainfall event, the CRSW of most layers in USR stand was not different from the other three stands, while the CRSW in the LIT and NTN stands was significantly higher than that in the HIT stand in the 0-100 cm soil layers, suggesting that soil in the LIT and NTN stands had a greater water-holding capacity than that in the HIT stand. In addition, soil properties were the main factors directly affecting the CRSW, explaining 60% and 37% of the variation in the CRSW on the first and seventh days after heavy rainfall, respectively. Overall, compared to the HIT stand, the LIT and NTN stands showed greater capacity in retaining rainwater. Therefore, under expected global changes with frequent occurrences of extreme precipitation events, methods involving light-intensity and no thinning should be employed to build up soil and water conservation functions, which will be critical for keeping water-holding capacity and moderating floods.

Potato leafroll virus (PLRV) and Potato virus Y (PVY) are two important viruses causing serious potato yield losses in the North-east region and other planting areas in India. As a consequence, it is urgent to develop an efficient and quick method for the identification and diagnosis in the field. The results presented here showed that the reverse transcription loop-mediated isothermal amplification (RT-LAMP) method was efficient and sensitive than reverse transcription-polymerase chain reaction (RT-PCR) for the detection of PLRV and PVY. The RT-LAMP primers specifically targeted PLRV and PVY (including PVY^O, PVY^N, and PVY^NTN strains) and resulted in typical sigmoidal amplification curves. Ten-fold serial dilutions of PLRV and PVY total RNA indicated that RT-LAMP is faster and at least a hundred times more sensitive than RT-PCR in detecting both the viruses. Additionally, samples that RT-PCR could not detect at a diluted concentration of 10^-3 and 10^-4 ng/μl were identified by RT-LAMP. Thus, RT-LAMP offers many advantages over RT-PCR such as low cost and high accuracy, sensitivity, and specificity for the rapid diagnosis of plant virus diseases. In conclusion, the results highlighted the efficacy of the RT-LAMP method in quickly detecting PLRV and PVY in infected plants.

Keywords: Potato; Potato leafroll virus; Potato virus Y; RT-LAMP; RT-PCR.

Background: Pregnant women, particularly in developing countries are facing a huge burden of preeclampsia (PE) leading to high morbidity and mortality rates. This is due to delayed diagnosis and unrecognised early targeted preventive measures. Adapting innovative solutions via shifting from delayed to early diagnosis of PE in the context of predictive diagnosis, targeted prevention and personalisation of medical care (PPPM/3 PM) is essential. The subjective assessment of suboptimal health status (SHS) and objective biomarkers of oxidative stress (OS) and angiogenic growth mediators (AGMs) could be used as new PPPM approach for PE; however, these factors have only been studied in isolation with no data on their combine assessment. This study profiled early gestational biomarkers of OS and AGMs as 3 PM approach to identify SHS pregnant mothers likely to develop PE specifically, early-onset PE (EO-PE) and late-onset PE (LO-PE).

Methods: A prospective cohort of 593 singleton normotensive pregnant (NTN-P) women were recruited at 10-20th (visit 1) and followed from 21 weeks gestation until the time of PE diagnosis and delivery. At visit 1, SHS was assessed using SHS questionnaire-25 (SHSQ-25) and women were classified as SHS and optimal health status (OHS). Biomarkers of OS (8-hydroxy-2-deoxyguanosine [8-OHdG], 8-epi-prostaglansinF2alpha [8-epi-PGF2α] and total antioxidant capacity [TAC]) and AGMs (vascular endothelial growth factor [VEGF-A], soluble Fms-like tyrosine kinase-1 [sFlt-1], placental growth factor [PlGF] and soluble endoglin [sEng]) were measured at visit 1 and time of PE diagnosis.

Results: Of the 593 mothers, 498 (248 SHS and 250 OHS) returned for delivery and were included in the final analysis. Fifty-six, 97 and 95 of the 248 SHS mothers developed EO-PE, LO-PE and NTN-P respectively, versus 14 EO-PE, 30 LO-PE and 206 NTN-P among the 250 OHS mothers. At the 10-20th week gestation, unbalanced levels of OS and AGMs were observed among SHS women who developed EO-PE than LO-PE compared to NTN-P women (p < 0.0001). The combined ratios of OS and AGMs, mainly the levels of 8-OHdG/PIGF ratio at 10-20th week gestation yielded the best area under the curve (AUC) and highest relative risk (RR) for predicting SHS-pregnant women who developed EO-PE (AUC = 0.93; RR = 6.5; p < 0.0001) and LO-PE (AUC = 0.88, RR = 4.4; p < 0.0001), as well as for OHS-pregnant women who developed EO-PE (AUC = 0.89, RR = 5.6; p < 0.0001) and LO-PE (AUC = 0.85; RR = 5.1; p < 0.0001).

Conclusion: Unlike OHS pregnant women, SHS pregnant women have high incidence of PE coupled with unbalanced levels of OS and AGMs at 10-20 weeks gestation. Combining early gestational profiling of OS and AGMs created an avenue for early differentiation of PE subtypes in the context of 3 PM care for mothers at high risk of PE.

Keywords: Angiogenic growth mediators; Oxidative stress; Predictive preventive personalised medicine (3P/PPPM); Preeclampsia; Risk assessment, Treatment algorithm, Patient stratification, Maternal health care, Public health education; Suboptimal health status.

Taspase1 is an Ntn-hydrolase overexpressed in primary human cancers, coordinating cancer cell proliferation, invasion, and metastasis. Loss of Taspase1 activity disrupts proliferation of human cancer cells in vitro and in mouse models of glioblastoma. Taspase1 is synthesized as an inactive proenzyme, becoming active upon intramolecular cleavage. The activation process changes the conformation of a long fragment at the C-terminus of the α subunit, for which no full-length structural information exists and whose function is poorly understood. We present a cloning strategy to generate a circularly permuted form of Taspase1 to determine the crystallographic structure of active Taspase1. We discovered that this region forms a long helix and is indispensable for the catalytic activity of Taspase1. Our study highlights the importance of this element for the enzymatic activity of Ntn-hydrolases, suggesting that it could be a potential target for the design of inhibitors with potential to be developed into anticancer therapeutics.

Keywords: Ntn-hydrolases; Taspase1; X-ray crystallography; anisotropy; cancer; intrinsically disordered proteins; plant-type L-asparaginases; proteases.

Human bone marrow mesenchymal stem cells (h-BMSCs) have the potential to differentiate into dopaminergic neuron-like cells to treat Parkinson's disease. The Notch signaling pathway has been implicated in the regulation of cell fate decisions such as differentiation of BMSCs. This study investigated changes in the expression of Notch-related genes in the differentiation of BMSCs in vitro into dopaminergic (DA) neuron-like cells. BMSCs transfected with empty lentiviral vectors served as the control group and those transfected with NTN and Lmx1α recombinant lentiviral vectors served as the experimental group. After induction and culture of NTN and Lmx1α-transfected h-BMSCs for 21 days, the cells exhibited features of dopaminergic neuron-like cells, which were observed by transmission and scanning electron microscopy and verified by immunofluorescence of tyrosine hydroxylase (TH) and dopamine transporter (DAT). These induced cells could secrete dopamine and had basic action potentials. Expression of the neural stem cell (NSC) markers, including octamer-binding protein (Oct4), paired box gene 6 (Pax6), and sex determining region Y-box 1 (SOX1), increased on day 14 of induction and decreased on day 21 of induction during differentiation. The human Notch signaling pathway PCR array showed a differential expression of Notch-related genes during the differentiation of h-BMSCs into DA neuron-like cells in vitro relative to that in the control group. In conclusion, h-BMSCs overexpressing NTN and Lmx1α can successfully be induced to differentiate into dopaminergic neuron-like cells with a neuronal phenotype exhibiting fundamental biological functions in vitro, and NTN and Lmx1α may affect the expression of Notch-related genes during differentiation.

Potato virus Y (PVY) belongs to the genus Potyvirus and is considered to be one of the most harmful and important plant pathogens. Its RNA-dependent RNA polymerase (RdRp) is known as nuclear inclusion protein b (NIb). The recent findings show that the genome of PVY replicates in the cytoplasm of the plant cell by binding the virus replication complex to the membranous structures of different organelles. In some potyviruses, NIb has been found to be localized in the nucleus and associated with the endoplasmic reticulum membranes. Moreover, NIb has been shown to interact with other host proteins that are particularly involved in promoting the virus infection cycle, such as the heat shock proteins (HSPs). HSP70 is the most conserved among the five major HSP families that are known to affect the plant-pathogen interactions. Some plant viruses can induce the production of HSP70 during the development of infection. To understand the molecular mechanisms underlying the interactive response to PVY^NTN (necrotic tuber necrosis strain of PVY), the present study focused on StHSC70-8 and PVY^NTN-NIb gene expression via localization of HSC70 and NIb proteins during compatible (susceptible) and incompatible (hypersensitive) potato-PVY^NTN interactions. Our results demonstrate that NIb and HSC70 are involved in the response to PVY^NTN infections and probably cooperate at some stages of the virus infection cycle. Enhanced deposition of HSC70 proteins during the infection cycle was associated with the dynamic induction of PVY^NTN-NIb gene expression and NIb localization during susceptible infections. In hypersensitive response (HR), a significant increase in HSC70 expression was observed up to 3 days post-inoculation (dpi) in the nucleus and chloroplasts. Thereafter, between 3 and 21 dpi, the deposition of NIb decreased, which can be attributed to a reduction in the levels of both virus accumulation and PVY^NTN-NIb gene expression. Therefore, we postulate that increase in the expression of both StHSC70-8 and PVY^NTN-NIb induces the PVY infection during susceptible infections. In contrast, during HRs, HSC70 cooperates with PVY^NTN only at the early stages of interaction and mediates the defense response signaling pathway at the later stages of infection.

Keywords: heat shock cognate 70 protein; hypersensitive response; nuclear inclusion b protein; plant–virus interaction.

Many intercellular communication processes, known as quorum sensing (QS), are regulated by the autoinducers N-acyl-l-homoserine lactones (AHLs) in Gram-negative bacteria. The inactivation of these QS processes using different quorum quenching (QQ) strategies, such as enzymatic degradation of the autoinducers or the receptor blocking with non-active analogs, could be the basis for the development of new antimicrobials. This study details the heterologous expression, purification, and characterization of a novel N-acylhomoserine lactone acylase from Actinoplanes utahensis NRRL 12052 (AuAHLA), which can hydrolyze different natural penicillins and N-acyl-homoserine lactones (with or without 3-oxo substitution), as well as synthesize them. Kinetic parameters for the hydrolysis of a broad range of substrates have shown that AuAHLA prefers penicillin V, followed by C₁₂-HSL. In addition, AuAHLA inhibits the production of violacein by Chromobacterium violaceum CV026, confirming its potential use as a QQ agent. Noteworthy, AuAHLA is also able to efficiently synthesize penicillin V, besides natural AHLs and phenoxyacetyl-homoserine lactone (POHL), a non-natural analog of AHLs that could be used to block QS receptors and inhibit signal of autoinducers, being the first reported AHL acylase capable of synthesizing AHLs.

Keywords: Actinoplanes utahensis; N-acyl-homoserine lactone; N-acyl-homoserine lactone acylase; Ntn-hydrolase; penicillin acylase; quorum quenching; quorum sensing.