Mucosal signatures of pathogenic T cells in HLA-B*27+ anterior uveitis and axial spondyloarthritis.

HLA-B*27 was one of the first HLA alleles associated with an autoimmune disease, i.e., axial spondyloarthritis (axSpA) and acute anterior uveitis (B27AAU), which cause joint and eye inflammation, respectively. Gastrointestinal inflammation has been suggested as a trigger of axSpA. We recently identified a bacterial peptide (YeiH) that can be presented by HLA-B*27 to expanded public T cell receptors (TCRs) in the joint in axSpA and the eye in B27AAU. While YeiH is present in enteric microbiota and pathogens, additional evidence that pathogenic T cells in HLA-B*27-associated autoimmunity may have had a prior antigenic encounter within the gastrointestinal tract remains lacking. Here, we analyze ocular, synovial, and blood T cells in B27AAU and axSpA, showing that YeiH-specific CD8 T cells express a mucosal gene set and surface proteins consistent with intestinal differentiation, including CD161, integrin α4ß7, and CCR6. In addition, we find an expansion of YeiH-specific CD8 T cells in the blood of axSpA and B27AAU over healthy controls, whereas influenza-specific CD8 T cells were equivalent across groups. Lastly, we demonstrate the dispensability of TRBV9 for antigen recognition. Collectively, our data suggest that, in HLA-B27-associated autoimmunity, early antigen exposure and differentiation of pathogenic CD8 T cells may occur in enteric organs.

Efficacy of WaveOne Gold and ProDesign RT systems in removing filling material: a micro-CT analysis.

The remaining filling material after retreatment can harbor bacteria and organic tissues that can influence the outcome of the therapy. AIM: The aim of this study was to evalúate, by micro-CT, the amount of filling material remaining in the root canal after its removal using WaveOne Gold or ProDesign RT. MATERIAL AND METHOD: Forty human mandibular canines were instrumented with the ProTaper Next system up to the X2 instrument (25.06) and filled with gutta-percha cones and AHPlus. Teeth were divided into 2 groups (n=20): WaveOne Gold 25.07 (WOG) and ProDesign RT 25.08 (PRT) for filling removal, after which they were scanned in a micro-CT device to quantify the volume of remaining filling material. The data were subjected to log 10 transformation, Student 's t-test was performed to account for multiple observationsper sample, significance was set at 5%. RESULTS: Student 's t-test showed that there was no difference between the two systems regarding the volume of remaining filling material in the thirds: apical (p = 0.392), middle (p = 0.065), or cervical (p = 0.918). CONCLUSIÓN: Remaining filling material was present in all groups and both systems were similar in removing root filling material in mandibular canines.

A permanencia de material obturador após o retratamento pode abrigar bactérias e tecidos orgánicos que podem influenciar o resultado da terapia. OBJETIVO: O objetivo deste estudo foi avaliar, por micro-CT, a quantidade de material obturador remanescente no canal radicular após a desobturagdo com WaveOne Gold e ProDesign RT. MATERIAL E MÉTODO: Quarenta caninos inferiores humanos foram instrumentados com o sistema ProTaper Next até o instrumento X2 (25.06) e obturados com cones de guta-percha e AHPlus. Os dentes foram divididos em dois grupos (n=20): WaveOne Gold 25.07 (WOG) e ProDesign RT 25.08 (PRT) e escaneados em micro-CT para quantificagdo do volume de material obturador remanescente. Os dados foram submetidos á transformando log 10, o teste t de Student foi realizado para contabilizar múltiplas observagoes por amostra, a significáncia foi fixada em 5%. RESULTADOS: O teste t de Student mostrou que ndo houve diferenga no volume de material obturador remanescente entre os dois sistemas nos tergos: apical (p = 0,392), médio (p = 0,065) ou cervical (p = 0,918). CONCLUSÃO: O material obturador remanescente estavapresente em todos os grupos e ambos os sistemas foram semelhantes na remogdo do material obturador radicular nos caninos inferiores.

Analysis of morphology and symmetry of the root canal system of incisors, premolars and mandibular molars using CBCT.

Knowledge of root canal internal anatomy and its variations is important forproper endodontic treatment. It is therefore necessary to investigate morphological aspects among different dental groups in the same patient to define the best protocol for the case. AIM: To evaluate the morphology and symmetry of homologous incisors, premolars and mandibular molars using cone beam computed tomography (CBCT). MATERIALS AND METHOD: Descriptive statistical analysis was performed for the frequency of categorical variables, and a chi-square test or Fisher 's exact test was used to test whether gender and side were associated with number of roots, number of canals, and Vertucci's classification. Forty-five CBCT scans were evaluated, and 444 mandibular teeth were analyzed. The number of roots, number of canals, classification of the canals in each root according to Vertucci and presence of a symmetrical relationship between pairs of posterior teeth were analyzed. RESULTS: The resuls showed that 74% of mandibular central incisors had type I root canal, 26% of mandibular lateral incisors had type I and, with a significant difference in the number of canals between males and females (p < 0.05). In mandibular first premolars, 70.5% had type I; and in mandibular second premolars, 98.5% had type I. Mandibular first molars had two roots in 98% of the cases. Second mandibular molars had two roots in 92.5% of the cases, one root in 6%, and three roots in 1.5%. Symmetry between central incisors was higher in females than in males. CONCLUSIÓN: Teeth of the same group can have different morphologies in the same patient.

0 conhecimento da anatomia interna e suas variagoes anatómicas é fator importante para o adequado tratamento endodóntico. Portanto, é necessário investigar esses aspectos morfológicos entre diferentes grupos dentários de um mesmo paciente para definir o melhor protocolo para o caso. OBJETIVO: Avaliar a morfologia e simetria de incisivos, pré-molares e molares inferiores homólogos por meio de tomografia computadorizada de feixe cónico (TCFC). MATERIAIS E MÉTODO: Foi realizada análise estatística descri-tiva para a frequéncia das variáveis categóricas e foi utilizado o teste do qui-quadrado ou teste exato de Fisher para testar a relagao entre sexo e lado em comparagao com número de raízes, número de canais e classificagao de Vertucci. Quarenta e cinco TCFC foram avaliadas e 444 dentes inferiores foram analisados. Foram considerados: o número de raízes, o número de canais, o tipo dos canais acordo com a classificagao de Vertucci e a presenga de relagao simétrica entre pares de dentes posteriores. RESULTADOS: Os resultados mostraram que 74% dos incisivos centrais inferiores tinham um canal radicular tipo 1 e 26% tinham dois canais; 73% dos incisivos laterais inferiores, 26%oeram do tipo I, tinham um canal e 27% tinham dois canais, com diferenga significativa no número de canais entre os grupos masculino e feminino (p < 0,05). Nos primeiros pré-molares inferiores, tipo I, um canal foi detectado em 70,5% e dois canais em 29,5%; nos segundos pré-molares inferiores, tipo I, um único canal foi detectado em 98,5%. O primeiro molar inferior foi observado com duas raízes em 98% e tres raízes em 2%o. O segundo molar inferior tinha duas raízes em 92,5% dos casos, uma raiz em 6% e tres raízes em 1,5%. A simetria foi maior nas mulheres em comparagao aos homens nos incisivos centrais. CONCLUSÃO: Pode-se concluir que dentes de um mesmo grupo podem apresentar morfologias diferentes no mesmo paciente.

Relationship between periapical lesions and sinus changes on multi-slice computed tomography sean.

Differentiating orofacial odontogenic pain/disorders from pain/disorders associated with maxillary sinusitis is important to avoid unnecessary dentalprocedures and to properly refer patients to colleagues/dentists and vice versa. AIM: To analyze the association between apical lesions and sinus changes and to evaluate the agreement between the diagnoses of an endodontist, a radiologist, an oral and maxillofacial surgeon, and an otolaryngologist. MATERIALS AND METHOD: 385 axial, coronal, and sagittal MSCT scans were selected using an image archiving andcommunication system (PACS). The examinations had been performed between 2018 and 2022. RESULTS: Apical lesions were observed in 36.10% of sinusitis cases, 73.8% of unilateralsinusitis cases, 48.7% of sinus floor discontinuity cases, and 67.2% of cases in which endodontic treatment had been performed. Agreement between the diagnoses made by the endodontist and those made by the other investigators was high for most study variables (k > 0.60). The exceptions were mucosal thickening, for which agreement between the endodontist and the other investigators was intermediate (k=0.397), and the presence of periapicallesions (k=0.010), previous endodontic treatment (k=0.013), and mucosal thickness (k=0.024), for which agreement between endodontists and radiologists was low. Conclusions: There was an association between sinus changes and apical lesions.

Diferenciar a dor/desordens odontogénicas orofaciais da dor/desordens as sociadas á sinusite maxilar é importante para evitar procedimentos odontológicos desnecessários e para encaminhar adequadamente os pacientes aos colegas/dentistas e vice-versa. OBJETIVO: Analisar a associagdo entre lesoes apicais e alteragóes sinusais e avaliar a concordáncia entre os diagnósticos de um endodontista, um radiologista, um cirurgido bucomaxilofacial e um otorrinolaringologista. MATERIAL E MÉTODO: foram avaliadas 385 imagens. RESULTADOS: Aslesoes apicais foram observadas em 36,10% dos casos de sinusite, em 73,8% dos casos de sinusite unilateral, em 48,7% dos casos de descontinuidade do assoalho do seio e em 67,2%odos casos em que o tratamento endodontico havia sido realizado. A concordancia entre os diagnósticos feitos pelo endodontista e os feitos pelos outros pesquisadores foi alta para a maioria das variáveis do estudo (k > 0,60). As excegoes foram o espessamento da mucosa, para o qual a concordáncia entre o endodontista e os outros pesquisadores foi intermediária (k=0,397) e a presenga delesoes periapicais (k=0,010), tratamento endodontico prévio (k=0,013) e espessura da mucosa (k=0,024), para os quais a concordáncia entre endodontistas e radiologistas foi baixa. CONCLUSÕES: Houve uma associagdo entre as alteragóes sinusais e aslesoes apicais.

Residential history, the electronic health record, and clinical medicine: A descriptive study in a lung transplant cohort.

Neighborhood level social determinants of health are commonly measured using a patient's most recent residential location. Not accounting for residential history, and therefore missing accumulated stressors from prior social vulnerabilities, could increase misclassification bias. We tested the hypothesis that the electronic health record could capture the residential history of lung transplant patients -a vulnerable population. After applying the Social Vulnerability Index (SVI) to individual residential histories, the most recent SVI equaled the first SVI in only 15.4% (58/374) of patients. There is a need for databases with residential histories to inform place-based determinants of health and applications to patient care.

The Near-Gapless Penicillium fuscoglaucum Genome Enables the Discovery of Lifestyle Features as an Emerging Post-Harvest Phytopathogen.

Penicillium spp. occupy many diverse biological niches that include plant pathogens, opportunistic human pathogens, saprophytes, indoor air contaminants, and those selected specifically for industrial applications to produce secondary metabolites and lifesaving antibiotics. Recent phylogenetic studies have established Penicillium fuscoglaucum as a synonym for Penicillium commune, which is an indoor air contaminant and toxin producer and can infect apple fruit during storage. During routine culturing on selective media in the lab, we obtained an isolate of P. fuscoglaucum Pf_T2 and sequenced its genome. The Pf_T2 genome is far superior to available genomic resources for the species. Our assembly exhibits a length of 35.1 Mb, a BUSCO score of 97.9% complete, and consists of five scaffolds/contigs representing the four expected chromosomes. It was determined that the Pf_T2 genome was colinear with a type specimen P. fuscoglaucum and contained a lineage-specific, intact cyclopiazonic acid (CPA) gene cluster. For comparison, a highly virulent postharvest apple pathogen, P. expansum strain TDL 12.1, was included and showed a similar growth pattern in culture to our Pf_T2 isolate but was far more aggressive in apple fruit than P. fuscoglaucum. The genome of Pf_T2 serves as a major improvement over existing resources, has superior annotation, and can inform forthcoming omics-based work and functional genetic studies to probe secondary metabolite production and disparities in aggressiveness during apple fruit decay.

Targeting lymphatic function in cardiovascular-kidney-metabolic syndrome: preclinical methods to analyze lymphatic function and therapeutic opportunities.

The lymphatic vascular system spans nearly every organ in the body and serves as an important network that maintains fluid, metabolite, and immune cell homeostasis. Recently, there has been a growing interest in the role of lymphatic biology in chronic disorders outside the realm of lymphatic abnormalities, lymphedema, or oncology, such as cardiovascular-kidney-metabolic syndrome (CKM). We propose that enhancing lymphatic function pharmacologically may be a novel and effective way to improve quality of life in patients with CKM syndrome by engaging multiple pathologies at once throughout the body. Several promising therapeutic targets that enhance lymphatic function have already been reported and may have clinical benefit. However, much remains unclear of the discreet ways the lymphatic vasculature interacts with CKM pathogenesis, and translation of these therapeutic targets to clinical development is challenging. Thus, the field must improve characterization of lymphatic function in preclinical mouse models of CKM syndrome to better understand molecular mechanisms of disease and uncover effective therapies.

Cis-regulatory evolution of the recently expanded Ly49 gene family.

Comparative genomics has revealed the rapid expansion of multiple gene families involved in immunity. Members within each gene family often evolved distinct roles in immunity. However, less is known about the evolution of their epigenome and cis-regulation. Here we systematically profile the epigenome of the recently expanded murine Ly49 gene family that mainly encode either inhibitory or activating surface receptors on natural killer cells. We identify a set of cis-regulatory elements (CREs) for activating Ly49 genes. In addition, we show that in mice, inhibitory and activating Ly49 genes are regulated by two separate sets of proximal CREs, likely resulting from lineage-specific losses of CRE activity. Furthermore, we find that some Ly49 genes are cross-regulated by the CREs of other Ly49 genes, suggesting that the Ly49 family has begun to evolve a concerted cis-regulatory mechanism. Collectively, we demonstrate the different modes of cis-regulatory evolution for a rapidly expanding gene family.

Expression of a single inhibitory Ly49 receptor is sufficient to license NK cells for effector functions.

Natural killer (NK) cells recognize target cells through germline-encoded activation and inhibitory receptors enabling effective immunity against viruses and cancer. The Ly49 receptor family in the mouse and killer immunoglobin-like receptor family in humans play a central role in NK cell immunity through recognition of MHC class I and related molecules. Functionally, these receptor families are involved in licensing and rejection of MHC-I-deficient cells through missing-self. The Ly49 family is highly polymorphic, making it challenging to detail the contributions of individual Ly49 receptors to NK cell function. Herein, we showed mice lacking expression of all Ly49s were unable to reject missing-self target cells in vivo, were defective in NK cell licensing, and displayed lower KLRG1 on the surface of NK cells. Expression of Ly49A alone on a H-2Dd background restored missing-self target cell rejection, NK cell licensing, and NK cell KLRG1 expression. Thus, a single inhibitory Ly49 receptor is sufficient to license NK cells and mediate missing-self in vivo.

Snyder-Robinson syndrome presenting with learning disability, epilepsy, and osteoporosis: a novel SMS gene variant.

Snyder-Robinson syndrome (SRS) is a rare X-linked recessive disorder characterized by a collection of clinical features including mild to severe intellectual disability, hypertonia, marfanoid habitus, facial asymmetry, osteoporosis, developmental delay and seizures. Whole genome sequencing (WGS) identified a mutation in the spermine synthase (SMS) gene (c.746 A>G, p.Tyr249Cys) in a male with kyphosis, seizures, and osteoporosis. His phenotype is unique in that he does not have intellectual disability (ID) but does have a mild learning disability. This case demonstrates a milder presentation of SRS and expands the phenotype beyond the reported literature.

Tenecteplase for Stroke at 4.5 to 24 Hours with Perfusion-Imaging Selection.

BACKGROUND: Thrombolytic agents, including tenecteplase, are generally used within 4.5 hours after the onset of stroke symptoms. Information on whether tenecteplase confers benefit beyond 4.5 hours is limited. METHODS: We conducted a multicenter, double-blind, randomized, placebo-controlled trial involving patients with ischemic stroke to compare tenecteplase (0.25 mg per kilogram of body weight, up to 25 mg) with placebo administered 4.5 to 24 hours after the time that the patient was last known to be well. Patients had to have evidence of occlusion of the middle cerebral artery or internal carotid artery and salvageable tissue as determined on perfusion imaging. The primary outcome was the ordinal score on the modified Rankin scale (range, 0 to 6, with higher scores indicating greater disability and a score of 6 indicating death) at day 90. Safety outcomes included death and symptomatic intracranial hemorrhage. RESULTS: The trial enrolled 458 patients, 77.3% of whom subsequently underwent thrombectomy; 228 patients were assigned to receive tenecteplase, and 230 to receive placebo. The median time between the time the patient was last known to be well and randomization was approximately 12 hours in the tenecteplase group and approximately 13 hours in the placebo group. The median score on the modified Rankin scale at 90 days was 3 in each group. The adjusted common odds ratio for the distribution of scores on the modified Rankin scale at 90 days for tenecteplase as compared with placebo was 1.13 (95% confidence interval, 0.82 to 1.57; P = 0.45). In the safety population, mortality at 90 days was 19.7% in the tenecteplase group and 18.2% in the placebo group, and the incidence of symptomatic intracranial hemorrhage was 3.2% and 2.3%, respectively. CONCLUSIONS: Tenecteplase therapy that was initiated 4.5 to 24 hours after stroke onset in patients with occlusions of the middle cerebral artery or internal carotid artery, most of whom had undergone endovascular thrombectomy, did not result in better clinical outcomes than those with placebo. The incidence of symptomatic intracerebral hemorrhage was similar in the two groups. (Funded by Genentech; TIMELESS ClinicalTrials.gov number, NCT03785678.).

Characterisation and engineering of a thermophilic RNA ligase from Palaeococcus pacificus.

RNA ligases are important enzymes in molecular biology and are highly useful for the manipulation and analysis of nucleic acids, including adapter ligation in next-generation sequencing of microRNAs. Thermophilic RNA ligases belonging to the RNA ligase 3 family are gaining attention for their use in molecular biology, for example a thermophilic RNA ligase from Methanobacterium thermoautotrophicum is commercially available for the adenylation of nucleic acids. Here we extensively characterise a newly identified RNA ligase from the thermophilic archaeon Palaeococcus pacificus (PpaRnl). PpaRnl exhibited significant substrate adenylation activity but low ligation activity across a range of oligonucleotide substrates. Mutation of Lys92 in motif I to alanine, resulted in an enzyme that lacked adenylation activity, but demonstrated improved ligation activity with pre-adenylated substrates (ATP-independent ligation). Subsequent structural characterisation revealed that in this mutant enzyme Lys238 was found in two alternate positions for coordination of the phosphate tail of ATP. In contrast mutation of Lys238 in motif V to glycine via structure-guided engineering enhanced ATP-dependent ligation activity via an arginine residue compensating for the absence of Lys238. Ligation activity for both mutations was higher than the wild-type, with activity observed across a range of oligonucleotide substrates with varying sequence and secondary structure.

Apixaban to Prevent Recurrence After Cryptogenic Stroke in Patients With Atrial Cardiopathy: The ARCADIA Randomized Clinical Trial.

Importance: Atrial cardiopathy is associated with stroke in the absence of clinically apparent atrial fibrillation. It is unknown whether anticoagulation, which has proven benefit in atrial fibrillation, prevents stroke in patients with atrial cardiopathy and no atrial fibrillation. Objective: To compare anticoagulation vs antiplatelet therapy for secondary stroke prevention in patients with cryptogenic stroke and evidence of atrial cardiopathy. Design, Setting, and Participants: Multicenter, double-blind, phase 3 randomized clinical trial of 1015 participants with cryptogenic stroke and evidence of atrial cardiopathy, defined as P-wave terminal force greater than 5000 µV × ms in electrocardiogram lead V1, serum N-terminal pro-B-type natriuretic peptide level greater than 250 pg/mL, or left atrial diameter index of 3 cm/m2 or greater on echocardiogram. Participants had no evidence of atrial fibrillation at the time of randomization. Enrollment and follow-up occurred from February 1, 2018, through February 28, 2023, at 185 sites in the National Institutes of Health StrokeNet and the Canadian Stroke Consortium. Interventions: Apixaban, 5 mg or 2.5 mg, twice daily (n = 507) vs aspirin, 81 mg, once daily (n = 508). Main Outcomes and Measures: The primary efficacy outcome in a time-to-event analysis was recurrent stroke. All participants, including those diagnosed with atrial fibrillation after randomization, were analyzed according to the groups to which they were randomized. The primary safety outcomes were symptomatic intracranial hemorrhage and other major hemorrhage. Results: With 1015 of the target 1100 participants enrolled and mean follow-up of 1.8 years, the trial was stopped for futility after a planned interim analysis. The mean (SD) age of participants was 68.0 (11.0) years, 54.3% were female, and 87.5% completed the full duration of follow-up. Recurrent stroke occurred in 40 patients in the apixaban group (annualized rate, 4.4%) and 40 patients in the aspirin group (annualized rate, 4.4%) (hazard ratio, 1.00 [95% CI, 0.64-1.55]). Symptomatic intracranial hemorrhage occurred in 0 patients taking apixaban and 7 patients taking aspirin (annualized rate, 1.1%). Other major hemorrhages occurred in 5 patients taking apixaban (annualized rate, 0.7%) and 5 patients taking aspirin (annualized rate, 0.8%) (hazard ratio, 1.02 [95% CI, 0.29-3.52]). Conclusions and Relevance: In patients with cryptogenic stroke and evidence of atrial cardiopathy without atrial fibrillation, apixaban did not significantly reduce recurrent stroke risk compared with aspirin. Trial Registration: ClinicalTrials.gov Identifier: NCT03192215.

Cranes soar on thermal updrafts behind cold fronts as they migrate across the sea.

Thermal soaring conditions above the sea have long been assumed absent or too weak for terrestrial migrating birds, forcing obligate soarers to take long detours and avoid sea-crossing, and facultative soarers to cross exclusively by costly flapping flight. Thus, while atmospheric convection does develop at sea and is used by some seabirds, it has been largely ignored in avian migration research. Here, we provide direct evidence for routine thermal soaring over open sea in the common crane, the heaviest facultative soarer known among terrestrial migrating birds. Using high-resolution biologging from 44 cranes tracked across their transcontinental migration over 4 years, we show that soaring performance was no different over sea than over land in mid-latitudes. Sea-soaring occurred predominantly in autumn when large water-air temperature difference followed mid-latitude cyclones. Our findings challenge a fundamental migration research paradigm and suggest that obligate soarers avoid sea-crossing not due to the absence or weakness of thermals but due to their low frequency, for which they cannot compensate with prolonged flapping. Conversely, facultative soarers other than cranes should also be able to use thermals over the sea. Marine cold air outbreaks, imperative to global energy budget and climate, may also be important for bird migration.

Network connectivity between psychological resilience and depression in prostate cancer patients.

OBJECTIVES: Prostate cancer (PCa) patients often experience depression. One possible buffer against stress-related depression is psychological resilience (PR), which has been described as heterogeneous in structure, like major depressive disorder (MDD). Although both of these constructs are central to understanding and assisting distressed PCa patients, no data have been reported on how they connect via network arrays at a component and symptom level. Such information has the potential to inform clinical practice with depressed PCa patients. METHODS: Using a cross-sectional design, 555 PCa patients completed the Patient Health Questionnaire-9 (PHQ-9) and the Connor-Davison Resilience Scale (CDRISC). Data were analysed via network analysis. RESULTS: Network analysis indicated that various CDRISC factors interacted with different PHQ-9 symptoms. For example, trust in one's instincts, tolerance of negative affect, and strengthening effects of stress (CDRISC) was associated with concentration problems and suicidal ideation (PHQ-9); positive acceptance of change, and secure relationships (CDRISC) was linked to low self-worth, anhedonia, fatigue/lethargy, motor problems, depressed mood, and concentration and appetite problems (PHQ-9). Similarly heterogeneous associations were found between individual CDRISC items and PHQ-9 symptoms. Network analytic figures depict both these sets of associations. CONCLUSIONS: As well as confirming the heterogeneous nature of PR and MDD in PCa patients, these findings argue for the further development of 'individualised' medicine approaches when working with PCa patients and their experiences of depression.

Mining the Penicillium expansum Genome for Virulence Genes: A Functional-Based Approach to Discover Novel Loci Mediating Blue Mold Decay of Apple Fruit.

Blue mold, a postharvest disease of pome fruits, is caused by the filamentous fungus Penicillium expansum. In addition to the economic losses caused by P. expansum, food safety can be compromised, as this pathogen is mycotoxigenic. In this study, forward and reverse genetic approaches were used to identify genes involved in blue mold infection in apple fruits. For this, we generated a random T-DNA insertional mutant library. A total of 448 transformants were generated and screened for the reduced decay phenotype on apples. Of these mutants, six (T-193, T-275, T-434, T-588, T-625, and T-711) were selected for continued studies and five unique genes were identified of interest. In addition, two deletion mutants (Δt-625 and Δt-588) and a knockdown strain (t-434KD) were generated for three loci. Data show that the ∆t-588 mutant phenocopied the T-DNA insertion mutant and had virulence penalties during apple fruit decay. We hypothesize that this locus encodes a glyoxalase due to bioinformatic predictions, thus contributing to reduced colony diameter when grown in methylglyoxal (MG). This work presents novel members of signaling networks and additional genetic factors that regulate fungal virulence in the blue mold fungus during apple fruit decay.

Avirulent Isolates of Penicillium chrysogenum to Control the Blue Mold of Apple Caused by P. expansum.

Blue mold is an economically significant postharvest disease of pome fruit that is primarily caused by Penicillium expansum. To manage this disease and sustain product quality, novel decay intervention strategies are needed that also maintain long-term efficacy. Biocontrol organisms and natural products are promising tools for managing postharvest diseases. Here, two Penicillium chrysogenum isolates, 404 and 413, were investigated as potential biocontrol agents against P. expansum in apple. Notably, 404 and 413 were non-pathogenic in apple, yet they grew vigorously in vitro when compared to the highly aggressive P. expansum R19 and Pe21 isolates. Whole-genome sequencing and species-specific barcoding identified both strains as P. chrysogenum. Each P. chrysogenum strain was inoculated in apple with the subsequent co-inoculation of R19 or Pe21 simultaneously, 3, or 7 days after prior inoculation with 404 or 413. The co-inoculation of these isolates showed reduced decay incidence and severity, with the most significant reduction from the longer establishment of P. chrysogenum. In vitro growth showed no antagonism between species, further suggesting competitive niche colonization as the mode of action for decay reduction. Both P. chrysogenum isolates had incomplete patulin gene clusters but tolerated patulin treatment. Finally, hygromycin resistance was observed for both P. chrysogenum isolates, yet they are not multiresistant to apple postharvest fungicides. Overall, we demonstrate the translative potential of P. chrysogenum to serve as an effective biocontrol agent against blue mold decay in apples, pending practical optimization and formulation.

Perinatal murine cytomegalovirus infection reshapes the transcriptional profile and functionality of NK cells.

Infections in early life can elicit substantially different immune responses and pathogenesis than infections in adulthood. Here, we investigate the consequences of murine cytomegalovirus infection in newborn mice on NK cells. We show that infection severely compromised NK cell maturation and functionality in newborns. This effect was not due to compromised virus control. Inflammatory responses to infection dysregulated the expression of major transcription factors governing NK cell fate, such as Eomes, resulting in impaired NK cell function. Most prominently, NK cells from perinatally infected mice have a diminished ability to produce IFN-γ due to the downregulation of long non-coding RNA Ifng-as1 expression. Moreover, the bone marrow's capacity to efficiently generate new NK cells is reduced, explaining the prolonged negative effects of perinatal infection on NK cells. This study demonstrates that viral infections in early life can profoundly impact NK cell biology, including long-lasting impairment in NK cell functionality.

NK cell expansion requires HuR and mediates control of solid tumors and long-term virus infection.

Natural killer (NK) cells are lymphocytes capable of controlling tumors and virus infections through direct lysis and cytokine production. While both T and NK cells expand and accumulate in affected tissues, the role of NK cell expansion in tumor and viral control is not well understood. Here, we show that posttranscriptional regulation by the RNA-binding protein HuR is essential for NK cell expansion without negatively affecting effector functions. HuR-deficient NK cells displayed defects in the metaphase of the cell cycle, including decreased expression and alternative splicing of Ska2, a component of the spindle and kinetochore complex. HuR-dependent NK cell expansion contributed to long-term cytomegalovirus control and facilitated control of subcutaneous tumors but not tumor metastases in two independent tumor models. These results show that posttranscriptional regulation by HuR specifically affects NK cell expansion, which is required for the control of long-term virus infection and solid tumors, but not acute infection or tumor metastases, highlighting fundamental differences with antigen-specific T cell control.