BACKGROUND There is increasing concern that a significant proportion of randomised controlled trials (RCTs) included in Cochrane reviews may not be trustworthy. Applying a trustworthiness screening tool (TST) has already had a clinically important effect on several reviews published by the Cochrane Pregnancy and Childbirth Group. OBJECTIVES We wanted to assess the impact of removing untrustworthy RCTs from already- published Cochrane reviews on a defined clinical area (ante- and post-natal nutritional interventions). METHODS We applied the tool to 18 Cochrane reviews (375 RCTs). The tool had four domains: i) is the research governance trustworthy; ii) are the baseline characteristics trustworthy; iii) is the study feasible; iv) are the results plausible?). When additional information was needed, authors were contacted using a standard template. At least two attempts were made to contact the authors. At the end of the evaluation process each study was classified as: i) included (YES to all domains); ii) excluded (retracted study); or iii) awaiting classification (any NO to the TST questions). RESULTS 95/375 studies (25%) were removed, affecting 14/18 (78%) reviews. 13/18 reviews (72%) showed a difference in the Summary of Findings tables (direction and size of effects and/or GRADE ratings). 6/18 Cochrane reviews (33%) were judged to require updating because of important differences in either in their conclusions, implication for practice, and/or implication for research. CONCLUSIONS Formal assessment of trustworthiness and inclusion only of studies that satisfy prespecified criteria for trustworthiness affect conclusions in a relatively large number of Cochrane reviews, with potentially important clinical implications for practice and research. The lack of consensus regarding the best tool(s) for assessing trustworthiness cannot be an excuse for ignoring this issue in future Cochrane reviews.

MAPK1 signaling pathway promotes development and survival of epithelial tissue. MAPKs family controls the activity of microenvironment by triggering the intracellular signaling. Present study was designed to understand the MAPK1 PPI by using STRING database and retrieve top twenty interaction network to analyze the significance of MAPK1 related proteins in lung Squamous cell carcinoma (SqCC). A cancer platform, cBioPortal was used to perform gene alteration analysis to uncover their participation in lung SqCC progression. TP53 gene showed high ratio alteration among eleven lung SqCC linked disease causing MAPK1 mediated pathway genes in comparative study of lung squamous cell carcinoma TCGA, nature 2012 dataset. In present study, using meta-analysis, it has been characterized that vital MAPK1 signaling pathway altered interactions that are MYLK, BCL2, TP53, BCL6, RAF1, CTTN, FGF19, FGF3, FGF4, ATR, TP63, HES1, CDKN2A, RASA, MRAS, TNFSF10, IL12A, TEK, PTEN, AGTR1, DVL3, EPHB1, TRIO, RPS6KA1, RPS6KA2, FOS, FGFR1, PAK2, PRKCI, PIK3CA, PIK3CG and KDR. These are involved in cell proliferation, growth, differentiation, tumor suppressed activities and apoptosis. In aberrant conditions they act as oncogenic and anti-apoptotic agents. The empirical validation of MAPK1 signaling cascade key interactions in lung SqCC may give fruitful selection of therapeutic targets for squamous cell carcinoma in future.

Background Eating disorders are prevalent in the adolescent and young adult population, with 2.7% of adolescents effected. The American Academy of Pediatrics recommends yearly screening for eating disorders in adolescents. Even with this recommendation, eating disorders often go underdiagnosed. AYAs with cancer possess several risk factors for eating disorders that may place them at an even higher risk, including receiving weight-altering therapies and having their weight/nutrition emphasized. Since these patients see their oncology team frequently, oncology clinics are opportune settings for eating disorder screening. This describes a single-institution study to implement screening for eating disorders in AYA patients in an oncology clinic. Procedures During regularly scheduled oncology visits, eligible patients were given the SCOFF questionnaire. Patients with an oncologic diagnosis aged 13 and older were screened. Patients with known eating disorders and patients receiving cytotoxic therapy were excluded. The questionnaire was scored by a study team member. Patients with a positive screening were referred to adolescent medicine. Results 163 eligible patients filled out the SCOFF questionnaire with 11 positive results (6.75%). Conclusions Our results demonstrate that eating disorder screening was successfully implemented in our pediatric oncology clinic. With a rate more than double than the general population, we observed that AYA patients with a history of cancer are indeed at a higher risk for eating disorders and should undergo routine screening. Since these patients have frequent oncology appointments, oncology clinics should implement screening for eating disorders. Further studies are needed to develop appropriate screening methods for on therapy patients.

This paper investigates the feasibility of using industrial waste acid to prepare geopolymer with significant environmental and economic benefits. Two types of waste acid were examined to discuss the effect of waste acid content on the microstructure, mechanical performance, thermal stability and heavy metal solidification efficiency. The core issue solved in this investigation is the harmless treatment of industrial waste acid and solidification efficiency of heavy metals, so as to reduce the pollution of heavy metals on the environment and the urgent demand for low carbon emissions in the construction and ceramic industries. The results showed that the metal elements (Ag, Fe, Mo, Cu and Ni) can be successfully solidified with over 95% solidification efficiency, indicating that the geopolymers are appropriate for hazardous material stabilisation, and the leaching of the heavy metals to the environment can be well controlled. The environmental and economic analysis indicates that the carbon and energy consumption intensity were reduced by 92.7% and 77.9%, respectively, compared to cement. What is more important is that the cost intensity is negative due to the income of waste acid treatment, meaning that additional value benefit was added to the product. This paves a promising way for the low-carbon transformation of ceramic enterprises and industrial waste treatment enterprises, drives further breakthroughs in high-tech industries, and promotes the sustainable development of the local national economy.

Saccharomyces cerevisiae, a generally recognized as safe (GRAS) microorganism, is regarded as an industrial workhorse for chemical and recombinant protein synthesis. In the biotechnological industry, multi-copy gene integration represents an effective strategy to maintain a high-level production of recombinant proteins-of interest, and to assemble multi-gene biochemical pathways. In the present study, we first created a copper-sensitive yeast strain by abolishing the CUP1 gene (encoding metallothionein that binds copper ions). Subsequently, we harnessed the CUP1 as a selection marker and combined the δ sites for multi-copy gene integration in S. cerevisiae. With the newly developed CUP1-tagging platform, multi-copy chromosomal integration of enhanced green fluorescent protein (eGFP) expression cassette was achieved, and the engineered strains remained quite stable after successive rounds of passaging. Taken together, we envision that the CUP1-tagging system would be a robust and useful method for protein overproduction and metabolic engineering applications in budding yeast.

Study 1 compared perceptions of female athletes in conservative or revealing uniforms. Participants (N=300) in a 2x3 experiment were randomly assigned to view images of three female lacrosse players wearing either tank tops or t-shirts and either short shorts, long shorts, or skirts. Participants rated the appropriateness of the uniform and the athletes’ capability, femininity, and objectification. A MANOVA revealed a significant main effect for shirt. The conservative shirt was seen as more appropriate than the revealing style; however, uniform type did not affect perceptions of the athletes. Study 2 measured how uniform styles are perceived by the women intended to wear them. Participants (N=156) were randomly assigned to view an image of either a revealing (racerback tank top and spandex shorts) or conservative (loose t-shirt and shorts) uniform and imagine themselves wearing it. Participants completed questions on their attitude toward the style, as well as their expected performance, self-objectification, and overall self-confidence. A MANOVA revealed a significant effect for uniform. Participants self-objectified more and were less confident in the revealing uniform despite preferring the style. Overall findings suggest that while uniforms do not affect others’ perceptions of female athletes, they do affect the athletes’ self-perceptions.

The study of the diet of extreme specialist species can help to understand the selective pressures that modulate the trophic strategies of raptors. This article provides insight into the trophic strategy of a little-known top predator and allows for a better understanding of the selective pressures that shape its diet. We studied the diet of European honey-buzzards, a raptor specialized in the consumption of social wasp brood. The ratio of predator to prey biomasses is one of the highest among raptors. We studied some factors that affect the energy demand of pairs which influence the diet composition and the daily rate of prey delivery to the nest. We explore hypotheses about the role of native and exotic vespids in the diet and the influence of the number and age of nestlings on diet composition, daily rate of prey delivery, and size of combs delivered. We installed trail cameras in 24 honey-buzzards nests in north-western Spain in 2018-2021. We estimated the proportion and daily rate of delivery of native common-wasps (Vespula vulgaris), invasive Asian-hornets (Vespa velutina), reptiles, and birds and the honey-buzzard´s preferences for vespid species. We performed LMMs, GLMMs, and MLRMs to analyse relationships between response variables and predictors. We identified 4611 prey. Honey-buzzards mainly consumed vespids (82% of prey). Common-wasps and Asian- hornets were almost the only two vespids consumed. The invasive hornet was the second most important prey consumed, but common-wasps were preferred. Vespids were more important as the age and number of nestlings increased. Our results suggest that the honey-buzzard´s diet is the adaptive result of the conflict between being a median-sized insect-eating migratory nidicolous raptor and collecting enough morsels for the growth of its nestlings, supporting the prey size and ingestion rate hypothesis. We discuss implications of our findings for the management of the invasive wasp

Muscle-specific kinase myasthenia gravis (MuSK MG) is caused by autoantibodies against MuSK in the neuromuscular junction (NMJ). MuSK MG patients have fluctuating, fatigable weakness, in particular of bulbar muscles. Severity differs greatly between patients, in spite of comparable autoantibody levels. One explanation for inter-patient and inter-muscle variability in sensitivity might be variations in compensatory muscle responses. Previously, we developed a passive transfer mouse model for MuSK MG. In preliminary ex vivo experiments we observed that muscle contraction, in particular of mice with milder myasthenia, had become partially insensitive to μ-Conotoxin-GIIIB, a blocker of skeletal muscle NaV1.4 voltage-gated sodium channels. We hypothesized that changes in NaV channel expression profile, possibly co-expression of (μ-Conotoxin-GIIIB insensitive) NaV1.5 type channels, might lower the muscle fibre’s firing threshold and facilitate neuromuscular synaptic transmission. To test this, we here performed passive transfer in mice, using ‘high’, ‘intermediate’ and ‘low’ dosing regimens of purified MuSK MG patient IgG4 and compared myasthenia levels, μ-Conotoxin-GIIIB resistance, muscle fibre action potential characteristics and firing thresholds. High- and intermediate-dosed mice showed clear, progressive myasthenia, not seen in low-dosed animals. However, diaphragm NMJ electrophysiology demonstrated almost equal myasthenic severities amongst all regimens. Nonetheless, low-dosed mouse diaphragms showed a much higher degree of μ-Conotoxin-GIIIB resistance. This was not explained by upregulation of Scn5a (the NaV1.5 gene), lowered muscle fibre firing thresholds or histologically detectable upregulated NaV1.5 channels. It remains to be established which factors are responsible for the μ-Conotoxin-GIIIB insensitivity and whether the NaV repertoire change is compensatory beneficial, or a bystander effect.

We compared voluntary drive and corticospinal responses during eccentric (ECC), isometric (ISO) and concentric (CON) muscle contractions to shed light on the mechanisms underpinning the greater force production despite lower electromyographic activity (EMG) commonly reported in ECC than ISO and CON. Sixteen participants (20-33 y) performed ISO and isokinetic (30˚/s) CON and ECC knee extensor contractions between 110˚ and 40˚ knee flexion, while EMG was recorded from vastus lateralis. Voluntary activation (VA) was measured during ISO, CON and ECC maximal voluntary contractions (MVCs). Transcranial magnetic stimulation elicited motor evoked potentials (MEPs) and corticospinal silent periods (CSP) during MVCs and submaximal contractions (30% maximum), and short-interval intracortical inhibition (SICI) in submaximal contractions. MVC torque was greater (P<0.01) during ECC (302.6±90.0 Nm) than ISO (269.8±81.5 Nm) and CON (235.4±78.6 Nm), but VA was lower (P<0.01) for ECC (68.4±14.9%) than ISO (78.3±13.1%) and CON (80.7±15.4%). Additionally, EMG/torque was lower (P<0.02) for ECC (1.9±1.1 μV.Nm-1) than ISO (2.2±1.2 μV.Nm-1) and CON (2.7±1.6 μV.Nm-1), CSP was shorter (p<0.04) for ECC (0.097±0.03 s) than ISO (0.109±0.02 s) and CON (0.109±0.03 s), and MEP amplitude was lower (P<0.01) for ECC (3.46±1.67 mV) than ISO (4.21±2.33 mV) and CON (4.01±2.06 mV). Similar results were found for EMG/torque and CSP during 30% contractions, but MEP amplitude and SICI showed no differences among contractions (p>0.05). Reduced VA during ECC may be at least partly explained by lower corticospinal excitability, while a shorter CSP may reflect extra muscle spindle excitation of the motoneurons from muscle lengthening, rather than reduced corticospinal inhibition.

The conversion of corn oil into quality methyl ester through the transesterification process is an age-old concept. Because of its higher degree of unsaturation, similar to soybean oil, transesterified corn oil can be used to produce bio-based resin. Replicating published methods did not result in the desired quantity and quality of corn methyl ester needed for bio-resin production, which requires further investigation. This research investigated the effect of neutralization on the yield and quality of methyl ester produced from corn oil. The reaction was conducted at 60 ºC with a methanol-oil ratio of 6:1 and NaOH as a catalyst. After stopping the reaction with HCl, a hypothesized recovered yield (>70%) of corn methyl ester was achieved. The process was conducted at different reaction times (0.5, 1, and 1.5 h) and acid amounts in neutralization (0, 25, 50, 75, and 100%). The methyl ester yield ranged from 45-79%. A statistical model was obtained with linear and quadratic terms, and the recovered yield varied significantly with the acid amount, reaction time, and their interactions. Stopping the reaction with 25% neutralization resulted in around 20% yield increase compared to no acid treatment. On the other hand, the yield was reduced by 18-24% with increasing time when no acid was added. All the measured characteristics of the produced corn methyl ester sample were found within the limits of ASTM D6751 pure methyl ester. Overall, the optimization of the neutralization process showed promise in increasing the yield of quality methyl ester from corn oil.

Accuracy of outcome definitions in Mendelian randomization of maternal healthQian Yang,1,2 Maria Carolina Borges1,2 1 MRC Integrative Epidemiology Unit at the University of Bristol, Bristol, UK2 Population Health Sciences, Bristol Medical School, University of Bristol, Bristol, UKDear Dr Papageorphiou,We recently read the article by Dr Ardissino and colleagues entitled ‘Genetically predicted body mass index and maternal outcomes of pregnancy: A two-sample Mendelian randomization study’ [1], where 11 outcomes were investigated. To conduct Mendelian randomization (MR) analyses, this study extracted associations of selected genetic variants with those outcomes from publicly available GWAS (genome-wide association study) summary data from FinnGen (the sixth release, total N=147,061 women) – a national-wide network of Finish Biobank [2]. We noticed that “postpartum depression” included in Ardissino et al was inconsistent with the commonly used definition of postnatal depression occurring within a year of delivery [3,4]. FinnGen defined this outcome based on the International Classification of Diseases 10th Revision (ICD-10) codes (ICD-10 F32, F33 and F53.0) among women with at least one episode of delivery (ICD-10 O80-O84), without considering the time interval between delivery and diagnosis of depression. Therefore, cases of “postpartum depression” could be ascertained at any time after giving birth and, therefore, could be unrelated to pregnancy. As a consequence, findings for “postpartum depression” in Ardissino et al should be interpreted with caution due to the unspecific outcome definition.The increasing availability of publicly available or accessible data from GWAS consortia (e.g. Early Growth Genetics) and large biobanks (e.g. UK Biobank and FinnGen), combined with the creation of automated pipelines (e.g. MR-Base [5] used by Ardissino et al), has supported an rapid increase in publications using two-sample MR. Though such a combination has great potential to promote open science and advance health research, including in maternal-child health, we cautioned that detailed understanding of procedures used to generate GWAS summary data underlying MR analyses is of major importance to obtain reliable evidence and interpretable findings.

Objective: Evaluate NHS-funded fertility treatment policies across England, Scotland, and Wales, assessing their alignment with the needs of those trying to conceive and comparing eligibility against NICE guidance. Design: Review of current NHS-funded fertility treatment policies and retrospective analysis of participant data. Setting: Not applicable. Population: Reproductive-age UK-based women using Hertility Health services from 2021-2023, focusing on those actively trying to conceive (TTC) or planning future pregnancies. Methods: Policy evaluation and data review without interventions. Main Outcome Measures: Eligibility for NHS-funded fertility treatment under UK regional policies versus NICE guidance. Results: In total, out of 2980 individuals actively trying to conceive and 6282 individuals planning for future pregnancies, 1700 (57.0%) and 2750 (43.8%) respectively would be ineligible for state-funded treatment under current regional policies: 1551 (57.7%) in England, 123 (53.2%) in Scotland, and 26 (41.9%) in Wales for those actively trying to conceive, and 2558 (43.9%), 146 (41.7%), and 46 (44.2%) for those planning future pregnancies respectively. In contrast, only 248 (8.3%) of those actively trying to conceive and 149 (2.4%) of those planning for future pregnancies would be ineligible if the NICE guidance were universally applied. Conclusions: By imposing outdated clinical and non-clinical qualifying criteria, the current NHS-funded fertility treatment policies exclude a substantial number of individuals requiring treatment. This necessitates a swift policy overhaul, as promised in the 2022 UK Women’s Health Strategy, to avoid further age-related fertility loss in women who require treatment imminently.

With compact footprint, low energy consumption, high scalability, and mass producibility, chip-scale integrated devices are an indispensable part of modern technological change and development. Recent advances in two-dimensional (2D) layered materials with their unique structures and distinctive properties have motivated their on-chip integration, yielding a variety of functional devices with superior performance and new features. To realize integrated devices incorporating 2D materials, it requires a diverse range of device fabrication techniques, which are of fundamental importance to achieve good performance and high reproducibility. This paper reviews the state-of-art fabrication techniques for the on-chip integration of 2D materials. First, an overview of the material properties and on-chip applications of 2D materials is provided. Second, different approaches used for integrating 2D materials on chips are comprehensively reviewed, which are categorized into material synthesis, on-chip transfer, film patterning, and property tuning / modification. Third, the methods for integrating 2D van der Waals heterostructures are also discussed and summarized. Finally, the current challenges and future perspectives are highlighted.

All-optical signal processing based on nonlinear optical devices is promising for ultrafast information processing in optical communication systems. Recent advances in two-dimensional (2D) layered materials with unique structures and distinctive properties have opened up new avenues for nonlinear optics and the fabrication of related devices with high performance. This paper reviews the recent advances in research on third-order optical nonlinearities of 2D materials, focus-ing on all-optical processing applications in the optical telecommunications band near 1550 nm. First, we provide an overview of the material properties of different 2D materials. Next, we review different methods for characterizing the third-order optical nonlinearities of 2D materials, including the Z-scan technique, third-harmonic generation (THG) measurement, and hybrid device characterization , together with a summary of the measured n2 values in the telecommunications band. Finally , the current challenges and future perspectives are discussed.

Microwave photonic (MWP) transversal signal processors offer a compelling solution for realizing versatile high-speed information processing by combining the advantages of reconfigurable electrical digital signal processing and high-bandwidth photonic processing. With the capability of generating a number of discrete wavelengths from micro-scale resonators, optical microcombs are powerful multi-wavelength sources for implementing MWP transversal signal processors with significantly reduced size, power consumption, and complexity. By using microcomb-based MWP transversal signal processors, a diverse range of signal processing functions have been demonstrated recently. In this paper we provide a detailed analysis for the errors induced by experimental imperfections processors. First, we investigate the errors arising from different sources including imperfections in the microcombs, the chirp of electro-optic modulators, chromatic dispersion of the dispersive module, shaping errors of the optical spectral shapers, and noise of the photodetector. Next, we provide a global picture quantifying the impact of error sources on the overall system performance. Finally, we introduce feedback control to compensate the errors caused by experimental imperfections, achieving significantly improved accuracy. These results provide a guide for optimizing the accuracy of microcomb-based MWP transversal signal processors. Index Terms-Microwave photonics, optical microcombs, optical signal processing.

Objective The aim of this study was to assess inter-rater reliability for components of the Visual-Numeric Endometriosis Scoring System (VNESS). VNESS is a novel system for describing surgical findings in each compartment of the pelvis in a way that is simple to use, visually intuitive and mirrors a laparoscopic image of the pelvis. Design Validation study using short surgical laparoscopic video clips. Setting Anonymized video clips of endometriosis surgery scored by 50 Gynaecologists from 12 countries. Population Gynaecologists as scores of short surgical clips selected from a series of endometriosis procedures performed between 2012 and 2022. Methods 50 gynaecologists scored 93 short surgical videoclips using VNESS. 4650 scores were compared against a reference score and analysis was performed to assess inter-rater reliability. Main outcome measures The outcome measures were percentage agreement between given and reference scores, as well as intra-class correlation coefficients (ICC), Cohen Kappa and Quadratic Weighted Kappa Coefficients were calculated to evaluate inter-rater reliability. Results The highest and lowest percentage agreement with the reference score was seen in VNESS 4 (full thickness disease, 97% perfect agreement) and VNESS 1 (superficial disease, 53% perfect agreement) respectively. The intraclass correlation coefficient showed strong inter-rater reliability for all VNESS compartments except the vagina. Conclusions This study suggests that VNESS has excellent inter-observer reliability. Correlation is stronger with more severe disease.

In the context of sensory processing, visual discrimination is a fundamental function that enables survival. Previous findings suggest that such discrimination function can be decoded from electroencephalographic brain responses, especially by using oscillation feature. However, how to evaluate the fast visual discrimination is still unclear. In this study, we hypothesize that brain’s oscillatory activity in a passive viewing condition can serve as a sensitive predictor of fast visual discrimination. A visual multi-feature paradigm which allowing investigation of several different change types was used to record both event-related potentials (ERPs) and behavioral responses. First, we investigated separating the behavioral hit rate as a function of reaction time (categorized from 200 ms to 1000 ms with step of 100 ms). In the subsequent step, we extract the slow theta component from ERP’s time frequency represents with time frequency principal component analysis (TF-PCA) and correlate its average power with behavioral performance. Our results showed that the significant detect window for different deviants’ level was from 400 to 600 ms, while the hit rates in such detect window showed a significant correlation with the averaged time frequency power in the slow theta band during 100-300 ms latency for the color and shape deviants. These findings suggest that the oscillation power, particularly in the slow theta range, of the brain responses is a predictor of fast visual discrimination.

The current knowledge on insects preying on fruits is limited, and some of the scarce existing data on fruit-associated insects are secluded within the host institutions. Consequently, their value is not fully realized. However, the integration and interlinking of historical biocollections data of plants, fruits, and insects, collected in Kenya, within a digital framework have not been fully exploited. This necessitates the need to enhance accessibility by consolidating the historical biodiversity data onto a unified platform. To address these gaps, this article presents a description of the development of a web-based platform for data sharing and integrating biodiversity historical data of wild plants, fruits, associated insects, and their molecular barcodes (WiPtFruIM) while leveraging data science technologies. The platform holds invaluable potential in fruit pest management, by providing information on potential biocontrol agents for fruit pests, which can function as a decision-making tool and fruit-pest ecological modeling. The platform is invaluable information to a worldwide community (such as researchers, classroom education, nature enthusiasts, fruit pest management, modeling, etc.) to make informed decisions and build innovative tools.

The auditory system has an amazing ability to rapidly encode auditory regularities. Evidence comes from the popular oddball paradigm, in which frequent (standard) sounds are occasionally exchanged for rare deviant sounds, which then elicit signs of prediction error based on their unexpectedness (e.g., MMN, P3a). Here, we examine the widely neglected characteristics of deviants being bearers of predictive information themselves: Naïve participants listened to sound sequence constructed according to a new, modified version of the oddball paradigm including two types of deviants that followed diametrically opposed rules: one deviant sound occurred mostly in pairs (repetition rule), the other deviant sound occurred mostly in isolation (non-repetition rule). Due to this manipulation, the sound following a first deviant (either the same deviant or a standard) was either predictable or unpredictable based on its conditional probability associated with the preceding deviant sound. Our behavioural results from an active deviant-detection task replicate previous findings that deviant repetition rules (based on conditional probability) can be extracted when behaviourally relevant. Our electrophysiological findings obtained in a passive-listening setting indicate that conditional probability also translates into differential processing at the P3a level. However, MMN was confined to global deviants and was not sensitive to conditional probability. This suggests that higher-level processing concerned with stimulus selection and/or evaluation (reflected in P3a) but not lower-level sensory processing (reflected in MMN) considers rarely encountered rules.

Rationale. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) can provide precise analysis of a protein’s conformational dynamics across varied states, such as heat-denatured vs. native protein structures, localizing regions that are specifically affected by such conditional changes. Maximizing protein sequence coverage provides high confidence that regions of interest were located by HDX-MS, but one challenge for complete sequence coverage is N-glycosylation sites. The deuteration of peptides post-translationally modified by asparagine-bound glycans (glycopeptides) has not always been identified in previous reports of HDX-MS analyses, causing significant sequence coverage gaps in heavily glycosylated proteins and uncertainty in structural dynamics in many regions throughout a glycoprotein. Methods. We detected deuterated glycopeptides with a Tribrid Orbitrap Eclipse mass spectrometer performing data-dependent acquisition. An MS scan was used to identify precursor ions, if high-energy collision-induced dissociation (HCD) MS/MS of the precursor indicated oxonium ions diagnostic for complex glycans then electron transfer low-energy collision-induced dissociation (EThcD) MS/MS scans of the precursor identified the modified asparagine residue and the glycan’s mass. As in traditional HDX-MS the identified glycopeptides were then analyzed at the MS level in samples labeled with D 2O. Results. We report HDX-MS analysis of the SARS-CoV-2 spike protein ectodomain in its trimeric pre-fusion form, which has 22 predicted N-glycosylation sites per monomer, with and without heat treatment. We identified glycopeptides and calculated their average isotopic mass shifts from deuteration. Inclusion of the deuterated glycopeptides increased sequence coverage of spike ectodomain from 76% to 84%, demonstrated that glycopeptides had been deuterated, and improved confidence in results localizing structural re-arrangements. Conclusion. Inclusion of deuterated glycopeptides improves the analysis of the conformational dynamics of glycoproteins such as viral surface antigens and cellular receptors.

Valsa canker is a fungal disease of apple and pear growing areas in China even Asia. Malectin like kinases take crucial roles in plant recognition of the pathogen-induced signals and subsequent activation of partially host immune responses. However, the role of MEDOS1 (MDS1) in plant immunity has not yet been extensively explored. In the Malus domestic genome, we found that the MdMDS1, a homologous gene of MDS1 in Arabidopsis, responded to Valsa mali ( Vm) and Valsa pyri ( Vp). Over-expression of MdMDS1 decreased Valsa canker resistance of Pyrus betulifolia ‘Duli-G03’ suspension cells, and apple and pear fruits. During suspension cells in response to Vp signals, up-regulation of MdMDS1 inhibited expression of defense-related genes but activated cell wall related genes. Among these, pectin methylesterase gene PbePME1 was robustly induced. Further analysis confirmed that PbePME1, a negative regulator of Valsa canker resistance, was indispensable for MdMDS1’s function by interaction. Our results provide a theoretical reference for the resistant breeding and comprehensive control of Valsa canker of both apple and pear.

Nucleotide-binding, leucine-rich repeat (NLR) genes play a pivotal role in shaping plant effector-triggered immunity in response to pathogen invasions. However, the mechanisms governing the expression and behavior of NLRs, particularly in the context of head-to-head NLR gene pairs, in the presence of pathogens, remain uncovered. In this study, we dissected the Pik-H4 promoter (P Pik-H4) at the TATA boxes and conducted an in-depth investigation into split promoter activity using Agro-infiltration assays. The segments spanning 593-1232 bp and 2016-2492 bp (starting from -1 bp of Pik1-H4) within P Pik-H4 emerged as core regions for expressing Pik1-H4 and Pik1-H4 respectively. Nevertheless, merging these two core fragments failed to recover the promoter activity in both directions. Employing Gus staining, promoter activity assays and qRT-PCR, we unveiled the co-expression of Pik1-H4 and Pik2-H4 throughout the whole plant. Additionally, in the presence of the rice blast fungus, their co-amplification was observed in leaves and leaf sheaths. Strikingly, Pik-H4 exhibited heightened expression within vascular bundles. Moreover, perturbing the Pik1-H4 and Pik2-H4 co-expression relationship via overexpression in rice or Nicotiana did not disrupt the immune response. Upon infection, the singleton Pik 1-H4 localized within vesicles, while Pik 2-H4 predominantly occupied the nucleus within leaf sheath cells. Transcriptome analysis highlighted Pik-H4-mediated resistance triggering a transcriptome reprogramming between 12 and 24 hours post-inoculation. Notably, overexpression of Pik1-H4 or Pik2-H4 enriches various pathways compared to the Pik-H4 Lijiangheituanxingu near-isogenic lines. In summary, these findings unravel the intricate dynamics of co-expression and singular functionality within NLR bidirectional gene pairs upon pathogen invasion.

Introduction Endometriosis is a chronic estrogen-dependent disease characterized by the ectopic presence of endometrial glands and stroma outside the uterine cavity, estimated to affect up to 10% of reproductive-aged women.1 Although the clinical picture of the disease is widely varied, chronic pelvic pain appears to be the most prevalent presentation among patients with endometriosis. Furthermore, endometriosis has been identified as the most common cause of secondary dysmenorrhea.2 While the contributing factors leading to endometriosis remain unclear, several risk factors have been highlighted in the literature suggested to play a role in the pathogenesis of endometriosis. A recent systematic review highlighted consumption of red meat, trans fatty acids (TFA), and saturated fatty acid (SFA) increase the risk of endometriosis, whereas a higher intake of total dairy [all low-fat and high-fat dairy foods] was connected with lower risk of developing endometriosis.3Similarly, vitamin D, calcium, and fruits and vegetables consumption are additional factors that led to a decreased risk of endometriosis.4,5 Interestingly, Asian women are more likely to develop endometriosis which suggests race\ethnicity may play a role in developing endometriosis, as well as disease severity.6Furthermore, lean body size, menarche before the age of 12 years, and short menstrual cycle length have been associated with a lower risk.7 Endometriosis can be broadly classified into three subtypes according to its pathophysiology and anatomical location: superficial endometriosis, ovarian endometrioma (OMA), and deep infiltrating endometriosis (DIE).8 Superficial endometriosis, also known as peritoneal endometriosis found in 15-50% of women diagnosed with endometriotic lesions.9 While ovarian Endometrioma, also known as chocolate cyst, is found in 50% of women complaining of infertility.10 Further, Deep infiltrating endometriosis consider the most severe form of endometriosis, with an estimated prevalence of 14–20% in women with endometriosis.11 Several options can be used for the management of endometriosis including medical, surgical, and assisted reproductive technology (ART). The treatment of choice depends on the patient’s condition, if the main complaint is chronic pain, analgesics, and hormonal treatments can be used.12 However, a large majority of endometriosis patients seeking treatment for infertility, surgery or medically assisted reproduction may be the optimal choice of management for endometriosis-associated infertility.13 In this review, we aim to compare and contrast the pathology of adenomyosis with endometriosis as well as investigate uterine disorders. In addition, understanding the unique aspects of endometriosis and adenomyosis may direct future research, leading to enhanced management plans and therapeutic techniques, and potentially novel therapeutic interventions. Thus, this presents an opportunity for researchers to advance medical knowledge and divulge new insights into the underlying etiology of uterine disorders.

A document by Sara Abou Al-Saud. Click on the document to view its contents.