The endophytic fungi of certain grasses and other herbaceous plants have long been known to provide plants with anti-herbivore defense compounds, but there is little information about whether the endophytes of trees also engage in such mutualisms. We investigated the influence of the endophytic fungus Cladosporium cladosporioides on the chemical defenses of black poplar (Populus nigra) trees and the consequences for feeding preference and fitness of herbivorous insects and insect community assembly. Endophyte colonization increased both constitutive- and induced poplar defenses. Generalist Lymantria dispar larvae preferred and performed better on uninfected over endophyte-infected poplar leaves, most likely due to higher concentrations of salicinoids in endophytic leaves and the endophyte-produced alkaloid stachydrine. Under field conditions, the endophytic fungus also shapes insect community assembly in young black poplar trees. Our results show that endophytic fungi can play a major role both in defending trees against herbivorous insects and in structuring insect communities.

Adrenal infarction with latent myelodysplastic/myeloproliferative neoplasm, unclassifiable with JAK2 V617F mutationShunichiro Yasuda1, Momoko Chiba1, Rie Nishitani2 and Takako Watanabe21Department of Hematology, Tokyo Kyosai Hospital, Tokyo, Japan2Department of Diabetes, Endocrinology and Metabolism, Tokyo Kyosai Hospital,Tokyo, JapanCorrespondence: Shunichiro Yasuda, M.D., Ph.D.Hematology Division Tokyo Kyosai Hospital 2-3-8 Nakameguro Meguro-ku Tokyo, 153-8934, JapanTel: +81-3-3712-3151 Fax 81-3-3793-5582E-mail:y-shunichiro@tkh.meguro.tokyo.jpMANUSCRIPT TYPE: Case reportAUTHOR CONTRIBUTIONSSY, MC, RN, TW: were involved in the patient care and management. SY, MC and RN: prepared the initial draft of the manuscript. SY, MC, TW: edited the draft and reviewed the manuscript. All authors approved the final version of the manuscript and agreed to be accountable for any aspects related to the accuracy or integrity of the work.ACKNOWLEDGMENTSThe authors would like to thank all clinicians involved in this case and Editage (www.editage.jp) for the English language editing.FUNDING INFROMATIONThere is no funding for this article.CONFLICT OF INTEREST STATEMENTThe authors have no conflicts of interest relevant to the content of this article.DATA AVAILABILITY STATEMENTThe data that support the findings of this study are available from the corresponding author upon reasonable request.ETHICS STATEMENTThis manuscript confirms to the provisions of the Declaration of Helsinki in 1995 (as revised in Brazil 2013).CONSENTWritten informed consent was obtained from the patient to publish this report in accordance with the journal‘s patient consent policy.Key Clinical MessageHematopoietic neoplasms are considered a differential diagnosis when encountering adrenal infarction. We should recognize thatJAK2 V617F mutation can cause thrombogenicity even if there are no abnormal findings in the blood cell counts.AbstractAdrenal infarction, a rare ailment, has been sporadically linked to hematopoietic neoplasms. A 46-year-old male encountered left adrenal infarction, which coincided with a progressive rise in platelet counts. Subsequent diagnosis revealed myelodysplastic/myeloproliferative neoplasm, unclassifiable (MDS/MPN-U), featuring a JAK2 V617F mutation. Simultaneously, the patient manifested multiple arteriovenous thromboses, necessitating treatment with edoxaban, aspirin, and hydroxyurea. Following thrombosis resolution, he was transferred to a transplantation center. This report delves into the thrombogenicity linked to the JAK2 V617F mutation, while also examining documented instances of adrenal infarction in myeloid neoplasms.Key words; adrenal infarction, myelodysplastic/myeloproliferative neoplasm, unclassifiable (MDS/MPN-U), JAK2 V617F, CHIPINTRODUCTIONAdrenal infarction is a rare condition that occasionally develops in a hypercoagulable state 1. Antiphospholipid antibody syndrome (APS), COVID-19 infection, and heparin-induced thrombocytopenia (HITT) have been reported as underlying diseases of adrenal infarction1-3. Adrenal infarctions have also been rarely linked to myeloproliferative neoplasms (MPN), such as polycythemia vera (PV) and essential thrombocythemia (ET) 4,5.Myelodysplastic/myeloproliferative neoplasm (MDS/MPN) is a hematological disorder that exhibits characteristics of both MDS and MPN6,7. According to the revised version of the 4th edition of the World Health Organization (WHO) classification, the categories of MDS/MPN include chronic myelomonocytic leukemia (CMML), juvenile myelomonocytic leukemia (JMML), atypical chronic myeloid leukemia (aCML), MDS/MPN with ring sideroblasts and thrombocytosis (MDS/MPN-RS-T), and MDS/MPN-unclassifiable (MDS/MPN-U)6-8. The diagnosis of MDS/MPN-U is based on ruling out any other subtype of MDS/MPN 7,8. Approximately 25% of cases with MDS/MPN-U have JAK2 V617F mutations8, which is a major mutation in patients with MPN9. As patients with MDS/MPN-U exhibit MPN characteristics, they may have a high risk of thrombosis. However, due to the rarity of this disease, the frequency of thrombosis in patients with MDS/MPN-U has not yet been thoroughly investigated.Herein, we present a case of JAK2 V617F-positive MDS/MPN-U with adrenal infarction. The present case had a unique clinical course in which the platelet count of the patient was almost normal at the time of adrenal infarction; however, later, the counts increased, and the characteristics of MPN became apparent. We report this case with a discussion of the thrombogenicity caused by the JAK2 V617F mutation and review previously reported cases of adrenal infarction in myeloid neoplasms.CASE HISTORY/ EXAMINATIONA 46-year-old previously healthy man presented to our emergency department with epigastric and left hypochondrial pain. He had no history of medication or family history of thrombophilia. On physical examination, his body temperature was 37.1 °C, blood pressure was 152/101 mmHg, heart rate was 118 bpm, and oxygen saturation was 97%. Electrocardiography (ECG) findings were normal, and chest radiograph showed no abnormalities. Blood tests revealed a slight increase in the platelet count (42.6×104/μL) and mild anemia (Hemoglobin: 12.9g/dL); the COVID-19 antigen test was negative. Contrast-enhanced computed tomography (CT) showed left adrenal hypertrophy and non-contrast-enhancing areas of the adrenal gland. Additionally, there was increased density of adipose tissue and fluid retention around the left adrenal gland (Fig.1A). Subsequently, he received a diagnosis of left adrenal infarction. The patient was urgently admitted to our hospital, and heparin 10000 U/day was initiated (Fig. 2). While the adrenal infarction was unilateral, there were no signs of adrenal insufficiency based on vital signs and laboratory data.DIFFERENTIAL DIAGNOSIS, INVESTIGATIONS AND TREATMENTDue to the potential for adrenal insufficiency, we initiated the administration of hydrocortisone as a steroid cover. Endocrine examinations indicated the maintenance of the adrenal function of the patient; consequently, the hydrocortisone dose was tapered and terminated. His epigastric and left hypochondrial pain steadily relieved, and follow-up contrast-enhanced CT on day 7 revealed resolution of the left adrenal infarction (Fig.1B). Subsequently, we transitioned the antithrombotic therapy from heparin 10000U/day to edoxaban 30 mg/day, and the patient was discharged on day 16. As for the thrombotic predisposition that led to the adrenal infarction, the increase in platelet counts was not remarkable. Both protein C activity and protein S activities were within the normal range(148% and 73%, respectively). The lupus anticoagulant was negative. Hence, we could not find any causes of adrenal infarction.OUTCOME AND FOLLOW-UPSoon after discharge from our hospital, the patient suffered from a headache. Considering the possibility of edoxaban-induced headache, edoxaban treatment was discontinued on day 20. Two months later, he presented to our hospital with a persistent headache despite discontinuing edoxaban treatment. He was diagnosed with cerebral venous thrombosis (CVT) on contrast-enhanced magnetic resonance imaging (MRI), and edoxaban treatment was resumed on day 88. Simultaneously, the patient presented with right hypochondrial pain, and contrast-enhanced CT showed mild hypertrophy of the right adrenal gland (Fig.1C). He was clinically diagnosed with right adrenal infarction. Furthermore, abdominal aortic mural thrombosis was incidentally detected on contrast-enhanced CT. Regarding multiple arteriovenous thromboses in a short period, his thrombotic predisposition was examined again. Blood tests showed that various autoantibodies were all negative, and the patient did not have diabetes or dyslipidemia (Table.1). However, the platelet counts, which were almost normal at the time of the first admission, gradually increased (Fig.2). Suspecting MPN, especially ET, we examined the presence of JAK2 V617F mutation, which was found to be positive. Bone marrow aspiration showed no increase in the blasts but showed dysplastic features in granulocytes, erythroblasts, and megakaryocytes; a chromosomal abnormality with der(1;7)(q10;p10) was found in 20 out of 20 cells. Bone marrow biopsy showed no bone marrow fibrosis. WT1mRNA was slightly high (71 copies/µgRNA). Since our patient had characteristics of both MDS and MPN and the diagnostic criteria for other MDS/MPN subtypes were not fulfilled, he was diagnosed with MDS/MPN-U. At the same time, the patient was aware of a rash resembling urticaria rash and pruritus on both lower legs, with mild increase in eosinophils (8.1%). The rash and pruritus improved with the use of an antihistamine cream. Since his thrombocytosis was not remarkable (59.7×10⁴/µL) and von Willebrand Factor (vWF) ristocetin cofactor activity (vWF:RCo) did not decrease (89%), the risk of bleeding might not be high 10,11. The patient was then treated with edoxaban (30 mg/day) and aspirin (100 mg/day).On day 131, the patient was admitted to our hospital again for close examination and treatment because his platelet counts further increased (Fig.2) and the development of a new thrombosis was expected. After admission, contrast-enhanced MRI and contrast-enhanced CT were performed, which showed resolution of all arteriovenous thromboses, including the right adrenal infarction (Fig.1D). The platelet counts rose to approximately 100×10⁴/µL; therefore, hydroxyurea 500 mg/day was started on day 137 as cytoreductive therapy, which was increased to 1000 mg/day on day 140. His eosinophil count also gradually increased with a skin rash flare-up. A skin biopsy was performed, leading to the pathological diagnosis of leukocytoclastic vasculitis, which was considered closely related to MDS with der(1;7)(q10;p10)10,11. After the hydroxyurea treatment, both the platelet and eosinophil counts decreased, leading to no recurrence of new thrombosis or improvement of the skin rash. Both JAK2 V617F allele burden and the copy numbers of WT1mRNA increased in our patient, suggesting that MDS/MPN-U progressed. Furthermore, MDS with der(1;7)(p10;q10) complicated by eosinophilia has aggressive clinical features and a poor prognosis 13,14. As the prognosis of allogeneic hematopoietic stem cell transplantation (allo-HSCT) is poor in patients with MDS/MPN-U who have progressive disease or severe complications 12, earlier allo-HSCT might be preferable. After obtaining adequate informed consent, including the risks and benefits of allo-HSCT, the patient was transferred to the transplantation center to receive allo-HSCT at the optimal time.DISCUSSIONIn this manuscript, we present a case of JAK2 V617F-positive MDS/MPN-U with multiple thromboses, including adrenal infarction. Although the precise mechanisms of thrombogenesis in MDS/MPN-U have not been elucidated, we speculate that MDS/MPN-U caused multiple thromboses since our patient did not have any other risk factors for thrombosis or a thrombotic predisposition. Notably, the first adrenal infarction occurred when the platelet count was only slightly elevated and MDS/MPN was not apparent. To date, various mechanisms of thrombogenesis byJAK2 V617F have been reported; 1) JAK2 V617F-positive neutrophils and monocytes release inflammatory cytokines, leading to arteriosclerosis and arterial thrombosis 13. 2)JAK2 V617F-positive neutrophils activate β1/2 integrin, promoting venous thrombosis 14,15. 3) Vascular endothelial cell expression of　JAK2 V617F promotes a prothrombotic state due to increased P-selectin expression 16. Recently, Clonal Hematopoiesis (CH) has been identified and genetic mutations associated with the disease have been detected before the development of hematological malignancies 17,18. In addition, when CH occurs and the variant allele frequency (VAF) exceeds 2%, it is called CH with indeterminate potential (CHIP) 17.JAK2 V617F-CHIP holders had a higher incidence of both arterial and venous thromboses than non-holders 17. In the present case, JAK2 V617F-CHIP may have been associated with the thrombogenicity of adrenal infarction.Previously, five cases of myeloid neoplasms that developed into adrenal infarctions have been reported (Table.2). Among these cases, two patients with ET had JAK2 V617F mutations 4,5. Indeed, one patient developed adrenal infarction before the diagnosis of ET, indicating that JAK2 V617F-CHIP was closely associated with adrenal infarction, as in our case 4. However, one patient with MDS/MPN-U did not harbor the JAK2 V617F mutation19, indicating that other gene mutations may contribute to the thrombogenesis in MDS/MPN-U. Furthermore, one case with MDS had adrenal infarction with no other thrombotic predisposition, such as MPN, suggesting that the hypercoagulable state associated with MDS may contribute to thrombogenesis 20. With reference to these case reports, other factors as well asJAK2 V617F mutation, may have caused the adrenal infarction in our case with MDS/MPN-U. A previous study suggested that SF3B1 may be a risk factor for thrombosis in MDS/MPN-RS-T, a subtype of MDS/MPN21. At present, SF3B1 mutation analysis is required for the accurate classification of MDS/MPN subtypes according to the latest 5th WHO classification. However, SF3B1 analysis can only be performed at a limited number of centers in Japan and is not available at our hospital. If SF3B1 or other gene mutation analyses become widespread and can be performed in daily practice, more accurate predictions can be made regarding the risk of thrombosis in MDS/MPN subtypes in the future.All six cases in Table.2, including the current case, developed complications such as adrenal insufficiency and thromboses at other sites. Among the six patients, five had bilateral adrenal infarction4,5,19,20,22 and four developed primary adrenal insufficiency that required steroid replacement therapies4,19,20,22. In our case, the adrenal infarction was unilateral and improved soon after edoxaban treatment. Although our patient subsequently developed adrenal infarction contralaterally, the lesions of adrenal infarction were small; therefore, the patient did not develop adrenal insufficiency.Among the six patients shown in Table.2, four developed thromboses at other sites, including one case of angina pectoris that required percutaneous coronary intervention 5. Our patient simultaneously developed CVT, abdominal aortic mural thrombosis, and adrenal infarction. In accordance with the recommendations of a previous study on the combination of antiplatelet and anticoagulant drugs for arteriovenous thrombosis in patients with MPN 23, the patient was treated with edoxaban and aspirin, and all thromboses disappeared.Although the six cases in Table.2 did not develop adrenal hemorrhagic infarction, they sometimes occurred after adrenal infarction. The mechanisms underlying adrenal infarction and adrenal hemorrhagic infarction are not fully understood; however, it is hypothesized that they are related to the unique vascular anatomy of the adrenal gland4,19,24. When treating adrenal infarction, attention should be paid to adrenal hemorrhagic infarction. In particular, patients with MPN have an increased risk of bleeding owing to a decrease in vWF activity associated with an increase in the platelet count25. Therefore, patients with adrenal infarction who have characteristics of MPN should be examined for vWF activity before starting treatment with antithrombotic agents.Patients with MDS/MPN-U generally have a poor prognosis with a median overall survival of 12.4 months, as reported by a previous study8. Although the efficacy of ruxolitinib or hypomethylating agents for MDS/MPN-U has been reported26,27, they are not curative, and the only curative treatment option is allo-HSCT 12. Since patients with severe complications before allo-HSCT have a worse prognosis, management of these complications is crucial for patients with MDS/MPN-U. Although our patient had multiple arteriovenous thromboses, they were managed with antithrombotic agents, and the patient was eventually transferred to the transplantation center without any complications.CONCLUSIONWe report a case of JAK2 V617F-positive MDS/MPN-U with adrenal infarction. Hematopoietic neoplasms are considered a differential diagnosis when encountering adrenal infarction with unknown causes. Even if there are no abnormal findings in the blood cell counts at the time of adrenal infarction, they should be carefully monitored, and when MPN is suspected, JAK2 V617F mutation should be examined. Finally, patients with adrenal infarction of hematopoietic neoplasms often have complications, and management of these complications is crucial, especially in patients eligible for allo-HSCT.REFERENCES1. Espinosa G, Santos E, Cervera R, et al. Adrenal involvement in the antiphospholipid syndrome: clinical and immunologic characteristics of 86 patients. Med (Baltim) . Mar 2003;82(2):106-18. doi:10.1097/00005792-200303000-000052. VanderVeer EA, Torbiak RP, Prebtani AP, Warkentin TE. Spontaneous heparin-induced thrombocytopenia syndrome presenting as bilateral adrenal infarction after knee arthroplasty. BMJ Case Rep . Dec 29 2019;12(12)doi:10.1136/bcr-2019-2327693. Leyendecker P, Ritter S, Riou M, et al. Acute adrenal infarction as an incidental CT finding and a potential prognosis factor in severe SARS-CoV-2 infection: a retrospective cohort analysis on 219 patients.Eur Radiol . Feb 2021;31(2):895-900. doi:10.1007/s00330-020-07226-54. Hada Y, Yamada A, Kobayashi T, et al. Bilateral Adrenal Infarction that Developed in Latent Essential Thrombocythemia. Intern Med . Jun 21 2023;doi:10.2169/internalmedicine.1947-235. Iemura T, Mano C, Oba A, et al. [Essential thrombocythemia accompanied by adrenal infarction]. Rinsho Ketsueki . 2019;60(2):106-111. doi:10.11406/rinketsu.60.1066. Palomo L, Meggendorfer M, Hutter S, et al. Molecular landscape and clonal architecture of adult myelodysplastic/myeloproliferative neoplasms. Blood . Oct 15 2020;136(16):1851-1862. doi:10.1182/blood.20190042297. Gerke MB, Christodoulou I, Karantanos T. Definitions, Biology, and Current Therapeutic Landscape of Myelodysplastic/Myeloproliferative Neoplasms. Cancers (Basel) . Jul 27 2023;15(15)doi:10.3390/cancers151538158. DiNardo CD, Daver N, Jain N, et al. Myelodysplastic/myeloproliferative neoplasms, unclassifiable (MDS/MPN, U): natural history and clinical outcome by treatment strategy.Leukemia . Apr 2014;28(4):958-61. doi:10.1038/leu.2014.89. Luque Paz D, Kralovics R, Skoda RC. Genetic basis and molecular profiling in myeloproliferative neoplasms. Blood . Apr 20 2023;141(16):1909-1921. doi:10.1182/blood.202201757810. Agha A, Bateman H, Sterrett A, Valeriano-Marcet J. Myelodysplasia and malignancy-associated vasculitis. Curr Rheumatol Rep . Dec 2012;14(6):526-31. doi:10.1007/s11926-012-0281-311. Komura A, Meguri Y, Matsubara C, et al. [Myelodysplastic syndrome with der (1;7)(q10;p10) complicated with eosinophilia and organizing pneumonia]. Rinsho Ketsueki . 2023;64(7):619-625. doi:10.11406/rinketsu.64.61912. Kurosawa S, Shimomura Y, Tachibana T, et al. Outcome of Allogeneic Hematopoietic Stem Cell Transplantation in Patients with Myelodysplastic/Myeloproliferative Neoplasms-Unclassifiable: A Retrospective Nationwide Study of the Japan Society for Hematopoietic Cell Transplantation. Biol Blood Marrow Transplant . Sep 2020;26(9):1607-1611. doi:10.1016/j.bbmt.2020.05.01313. Perner F, Perner C, Ernst T, Heidel FH. Roles of JAK2 in aging, inflammation, hematopoiesis and malignant Transformation. Cells . Aug 8 2019;8(8)doi:10.3390/cells808085414. Gupta N, Edelmann B, Schnoeder TM, et al. JAK2-V617F activates β1-integrin-mediated adhesion of granulocytes to vascular cell adhesion molecule 1. Leukemia . May 2017;31(5):1223-1226. doi:10.1038/leu.2017.2615. Edelmann B, Gupta N, Schnoeder TM, et al. JAK2-V617F promotes venous thrombosis through β1/β2 integrin activation. J Clin Invest . Oct 1 2018;128(10):4359-4371. doi:10.1172/jci9031216. Guy A, Gourdou-Latyszenok V, Le Lay N, et al. Vascular endothelial cell expression of JAK2(V617F) is sufficient to promote a pro-thrombotic state due to increased P-selectin expression. Haematologica . Jan 2019;104(1):70-81. doi:10.3324/haematol.2018.19532117. Misaka T, Kimishima Y, Yokokawa T, Ikeda K, Takeishi Y. Clonal hematopoiesis and cardiovascular diseases: role of JAK2V617F. J Cardiol . Jan 2023;81(1):3-9. doi:10.1016/j.jjcc.2022.02.00118. Yokokawa T, Misaka T, Kimishima Y, et al. Clonal hematopoiesis and JAK2V617F mutations in patients with cardiovascular disease. JACC CardioOncol . Mar 2021;3(1):134-136. doi:10.1016/j.jaccao.2021.01.00119. Hoshino Y, Manaka K, Sato J, et al. Recurrent bilateral adrenal infarction with myelodysplastic/myeloproliferative neoplasm-unclassifiable (MDS/MPN-U): a case report. BMC Endocr Disord . Jun 5 2023;23(1):128. doi:10.1186/s12902-023-01384-520. Lockett HA, Hamilton-Wood C, Vaidya B. Addison’s disease due to bilateral adrenal infarction in a patient with myelodysplastic Syndrome.Eur J GEN Med . 2011;8:72-74.21. Nathan DI, Feld J, El Jamal SM, Mascarenhas J, Tremblay D. Myelodysplastic syndrome/myeloproliferative neoplasm with ring sideroblasts and thrombocytosis: Ringing in a new future. Leuk Res . Apr 2022;115:106820. doi:10.1016/j.leukres.2022.10682022. Hirata R, Tago M, Yamashita S, et al. Acute abdominal pain due to atypical bilateral adrenal infarction in acute myeloid leukemia with alterations related to myelodysplasia: A case report. Clin Case Rep . Oct 2023;11(10):e7925. doi:10.1002/ccr3.792523. Tefferi A, Barbui T. Polycythemia vera and essential thrombocythemia: 2021 update on diagnosis, risk-stratification and management. Am J Hematol . Dec 2020;95(12):1599-1613. doi:10.1002/ajh.2600824. Michiels JJ, Berneman Z, Schroyens W, Krestin GP. Bilateral adrenal swelling as a cause of chest, back, and upper abdominal pain in essential thrombocythemia and polycythemia vera is due to microvascular ischemic thrombosis rather than to hemorrhage. Ann Hematol . Dec 2002;81(12):691-4. doi:10.1007/s00277-002-0500-525. Awada H, Voso MT, Guglielmelli P, Gurnari C. Essential thrombocythemia and acquired von Willebrand syndrome: The shadowlands between thrombosis and bleeding. Cancers (Basel) . Jun 30 2020;12(7)doi:10.3390/cancers1207174626. Al-Kali A, Abou Hussein AK, Patnaik M, et al. Hypomethylating agents (HMAs) effect on myelodysplastic/myeloproliferative neoplasm unclassifiable (MDS/MPN-U): single institution experience. Leuk Lymphoma . Nov 2018;59(11):2737-2739. doi:10.1080/10428194.2018.143670527. Wang Q, Dai HP, Liu DD, et al. Efficacy of ruxolitinib in a patient with myelodysplastic/myeloproliferative neoplasm unclassifiable and co-mutated JAK2, SF3B1 and TP53. Leuk Res Rep . 2020;14:100229. doi:10.1016/j.lrr.2020.100229Figure legendsFigure 1. Contrast-enhanced CT showing development (A) and improvement of (B) left adrenal infarction. Contrast-enhanced CT showing development (C) and improvement (D) of right adrenal infarction.A) The arrow shows left adrenal hypertrophy and non-contrast-enhanced areas, and arrowheads show increased lipid concentrations and fluid retention around the left adrenal gland. The patient was diagnosed with left adrenal infarction. B) The left adrenal infarction improved. C) The arrow indicates mild hypertrophy of the right adrenal gland. He was diagnosed with a right adrenal infarction. D) The right adrenal infarction improved.Figure 2. Clinical course of adrenal infarction withJAK2 V617F-positive MDS/MPN-U with der(1;7)(p10;q10).Plt, platelet; Eo, eosinophil; vWF:Rco, von Willebrand Factor ristocetin cofactor activity;U, unit; Lt, left; Rt, right; AI, adrenal infarction; CVT, cerebral venous thrombosis; AMT, aortic mural thrombosis. BMA, bone marrow aspiration; BMB, bone marrow biopsy

Botryllus schlosseri, is a model marine invertebrate for studying immunity, regeneration, and stress-induced evolution. Conditions for validating its predicted proteome were optimized using nanoElute® 2 deep-coverage LCMS, revealing up to 4,930 protein groups and 20,984 unique peptides per sample. Spectral libraries were generated and filtered to remove interferences, low-quality transitions, and only retain proteins with >3 unique peptides. The resulting DIA assay library enabled label-free quantitation of 3,426 protein groups represented by 22,593 unique peptides. Quantitative comparisons of a laboratory-raised with two field-collected populations revealed (1) a more unique proteome in the laboratory-raised population, and (2) proteins with high/low individual variabilities in each population. DNA repair/replication, ion transport, and intracellular signaling processes were unique in laboratory-cultured colonies. Spliceosome and Wnt signaling proteins were the least variable (highly functionally constrained) in all populations. In conclusion, we present the first colonial tunicate’s deep quantitative proteome analysis, identifying functional protein clusters associated with laboratory conditions, different habitats, and strong versus relaxed abundance constraints. These results empower research on B. schlosseri with proteomics resources and enable quantitative molecular phenotyping of changes associated with transfer from in situ to ex situ and from in vivo to in vitro culture conditions.

Skin microbiomes provide vital functions, yet knowledge about their species assemblages is limited - especially for non-model organisms. In this study, we conducted in situ manipulations and repeated sampling on wild-caught individuals of Rutilus rutilus. Treatments included translocation between fresh and brackish water habitats to investigate the role of environment; community rebooting by disinfection to infer host-microbe interactions; and housing in pairs to study the role of inter-host dispersal for the structure of microbiomes colonizing animals. Results revealed that fish skin microbiomes were biodiversity hotspots with highly dynamic composition that were distinct from bacterioplankton communities. External environmental conditions and individual-specific factors jointly determined the colonization-extinction dynamics, whereas inter-host dispersal had negligible effects. The dynamics of the microbiome composition was seemingly non-affected by reboot treatment, pointing to high resilience to disturbance in these microbial communities. Together, the manipulations demonstrate that host individual characteristics and environment interactively shapes the skin microbiome of fish. The results emphasize the role of inter-individual variability for the unexplained variation found in many host-microbiome systems, although the mechanistic underpinnings remain to be identified.

FEV 1/FVC normally decreases through childhood, increases briefly during early adolescence, and then declines throughout life. The physiology behind this temporary increase during early adolescence is not well understood. The objective of this study was to determine if this pattern also occurs in children with asthma. Design: Single-center, cross-sectional, retrospective analysis of pulmonary function tests (PFTs) obtained over a 5-year period in children 5 to 18 years of age with persistent asthma. Results: 1,793 patients satisfied all inclusion and exclusion criteria. Mean age (± SD) was 10.4 ± 3.8 years. 48% were female. Mean FEV 1 /FVC was 0.83 ± .09. FEV 1 /FVC in children with persistent asthma declined from age 5 to age 11 by 5.7 % compared to 7.3% in healthy girls, and 5.8% compared to 9.4% in healthy boys. FEV 1/FVC increased by 1.2% until age 16 in children with asthma, compared to 2.2% in healthy girls, and 2.5% compared to 2.3% in healthy boys. The ratio was lower in obese children with asthma at all ages but demonstrated the same curvilinear shape as in healthy children. In absolute terms, FEV 1 grew proportionately more than FVC during early adolescence, so the ratio of FEV 1/FVC increased during that period. The curvilinear shape of the curve remained in postbronchodilator testing, though significantly blunted. Conclusions: The “Shepherd’s Hook” pattern in the FEV 1/FVC curve is preserved in children with persistent asthma. This was also true in obese patients with asthma, although their FEV 1/FVC ratios were lower throughout all stages of childhood and adolescence.

1. The overwhelming majority of patients referred into secondary care with suspected head and neck cancer (HNC) do not have cancer (~95%). 2. During the COVID-19 pandemic telephone triage of patients with suspected HNC was necessary. During this time, a validated HNC risk-calculator, HaNC-RC-v2 (a set of symptomatology-based questions) was recommended by ENT UK to stratify patients into high or low risk of having HNC via telephone triage [(1)](#ref-0001) 3. Ufonia, a digital health company which uses an Artificial Intelligence (AI) voice assistant to automate clinical conversations via telephone, and *INSTITUTION*, were awarded an SBRI Healthcare grant to help develop an AI-delivered HNC triage telephone call. This was based on the HaNC-RC-v2 and co-created with HNC patients from the Heads2Gether charity via round-table discussions and one-to-one sessions. 4. Twenty-nine patients underwent a clinician-supervised AI-delivered HNC triage conversation as part of their standard telephone consultation. 100% of calls were completed with an average agreement of 89% between the clinician and the AI system for all symptoms asked. The technology was highly acceptable to patients with a median net promoter score (NPS) score of 8 out of 10. 5. Novel technologies involving AI automated telephone calls can be generated to remotely triage suspected HNC patients. This technology may offer an exciting opportunity to help departments triage suspected HNC referrals in an ever increasingly resource pressurised NHS.

A document by Naveen Gautam. Click on the document to view its contents.

Introduction:Sickle cell disease (SCD) is a group of inherited diseases characterised by the presence of haemoglobin S (Hb S), either by homozygosity for the sickle mutation (Hb SS) or by mixed heterozygosity with another beta-globin variant (e.g. sickle beta thalassaemia, Hb SC disease). It is one of the most common haemoglobinopathies, especially in Middle Eastern countries. The hallmarks of SCD are recurrent vaso-occlusive crises and haemolytic anaemia. Vaso-occlusive crises manifest as acute pain crises, acute chest syndrome and priapism - they are mainly caused by obstructions of the microcirculation and lead to tissue hypoxia and severe pain.[1][2][3]Priapism is defined as a persistent erection of the penis that is not associated with sexual interest or desire. It can occur with low flow (ischaemic, vaso-occlusive) or high flow (non-ischaemic). In some series, it is estimated that 35 to 45 per cent of men with sickle cell disease (SCD) are affected by priapism. Ischaemic episodes lasting ≥4 hours (severe episodes) are of particular concern as they carry a high risk of permanent tissue damage from penile compartment syndrome. There is another vriant of ischaemic priapism, known as stuttering priapism, which is characterised by brief, recurrent episodes of transient, self-limited priapism.[4][5]Recent therapeutic advances have seen the approval of crizanlizumab, a monoclonal antibody targeting P-selectin, by the FDA in 2019 for reducing the frequency of SCD-related vaso-occlusive crises. Nonetheless, the efficacy of crizanlizumab in managing SCD-associated priapism has yet to be thoroughly investigated, underscoring a vital area for further clinical exploration [6].

Changes in leaf temperature are known to drive stomatal responses, because the leaf-to-air water vapor gradient (Δ w) increases with temperature if ambient vapor pressure is held constant, and stomata respond to changes in Δ w. However, the direct response of stomata to temperature (DRST; the response when Δ w is held constant by adjusting ambient humidity) has been examined far less extensively. Though the meager available data suggest the response is usually positive, results differ widely and defy broad generalization. As a result, little is known about the DRST. This review discusses the current state of knowledge about the DRST, including numerous hypothesized biophysical mechanisms, potential implications of the response for plant adaptation, and possible impacts of the DRST on plant-atmosphere carbon and water exchange in a changing climate.

Semi-arid alpine ecosystems on the Tibetan Plateau are experiencing rapid climate change and extensive anthropogenic activity, concomitant with the encroachment of shrubs. Shrub encroachment leads to changes in both the structure and functioning of semi-arid alpine steppes. Yet, the encroaching impacts of shrubs on the above- and belowground ecosystem functioning of the semi-arid alpine steppes remains uncertain. To quantify shrub encroachment impacts on ecosystem functioning of the semi-arid alpine steppes, two shrub encroached alpine steppe sites are investigated, with one site encroached by a leguminous shrub ( Caragana spinifera) and another site encroached by a non-leguminous shrub ( Dasiphora fruticosa). Results showed that following both Leguminosae and Non-Leguminosae shrub encroachment, not only do these alpine steppes greatly increase the individual ecosystem functions but that this included significant enhancing of the ecosystem multifunctionality (EMF). We concluded that the main impacts of shrub encroachment were to facilitate belowground EMF (BEMF) rather than aboveground EMF (AEMF) in alpine steppes. Our findings also highlight that soil nutrients play critical roles in driving ecosystem functioning responses to shrubs encroachment. These findings further our understanding of shrub encroachment impacts on ecosystem functioning of the Tibetan semi-arid alpine steppes.

Duddingtonia flagrans, a nematode-eating fungus, is an effective component of animal parasitic nematode biocontrol agents. In the dried formulation, the majority of spores are in an endogenous dormant state. This study focuses on dormant chlamydospore and non-dormant chlamydospore of D. flagrans to investigate the differences in cAMP and protein content between the two types of spores. In this study, cAMP and soluble proteins were extracted from the non-dormant chlamydospore and dormant chlamydospore of D. flagrans isolates SDH035 and DH055, respectively. The cAMP Direct Immunoassay Kit and Bradford protein concentration assay kit (Coomassie brilliant blue method) were used to detect the cAMP and protein content in two types of spores. Results showed that the content of cAMP in dormant spores of both isolates was significantly higher than that in non-dormant spores (p<0.05). The protein content of dormant spores in DH055 bacteria was significantly higher than that of non-dormant spores (p<0.05). In addition, the protein content of dormant spores of the SDH035 strain was slightly higher than that of non-dormant spores, but the difference was not significant (p>0.05). The results obtained in this study provide evidence for the biochemical mechanism of chlamydospore dormancy or the germination of the nematophagous fungus D. flagrans.

The idea of a Cyber-Physical-Social System, or CPSS for short, is a relatively new concept that has emerged as a response to the requirement to comprehend the influence that Cyber-Physical Systems (CPS) have on people and vice versa. Conversational assistants (CAs), also called bots, are dedicated to oral or written communication. Over time, the CAs have gradually diversified to today touch various fields such as e-commerce, healthcare, tourism, fashion, travel, and many others sectors. Natural-language understanding (NLU) is fundamental in the Natural Language Processing (NLP) field. Identifying user intents from natural language utterances is a crucial step in conversational systems, and the diversity in user utterances makes intent detection even a challenging problem. Recently, with the emergence of Deep Neural Networks. New State of the Art (SOA) results have been achieved for different NLP tasks. Recurrent Neural networks (RNNs) and recent Transformer architectures are two major players in those improvements. In addition, RNNs have been playing an increasingly important role in sequence modeling in different application areas. On the other hand, Transformer models are new architectures that benefit from the attention mechanism, extensive training datasets, and compute power. First, this review paper presents a comprehensive overview of RNN and Transformer models. Then, a comparative study of the performance of different RNNs and Transformer architectures for the specific task of intent recognition for CAs which is a fundamental task of NLU.

Real-time ultrasound guided segmental bronchoscopic insufflation in a Tay-Sachs patient with atelectasisMatej Šapina, MD, PhD1,2,3*, Bojana Olujić, MD1,2, Tihana Nađ, MD1,2, Hrvoje Vinković, MD1,2, Zdravka Krivdić Dupan, MD1,2, Nikolina Hamidović, MD1, Mia Damašek, MD1,2, Krešimir Milas, MD1, Željko Zubčić, MD, PhD1,2, and Ivan Erić, MD, PhD1,21University hospital center Osijek, 31000 Osijek, Croatia2Medical faculty Osijek, 31000 Osijek, Croatia3Faculty of dental medicine and health Osijek, 31000 Osijek, Croatia*Corresponding author: sapina.matej@yahoo.comPhone: +385 31 512 279Keywords: bronchoscopy, atelectasis, Tay-Sachs diseaseIntroductionTo the Editor,Atelectasis is the collapse of lung tissue and is a common problem in patients suffering from severe neurological diseases. Treatment depends on the underlying cause, and ranges from respiratory physiotherapy, mucus clearance methods, and in some cases bronchoscopic interventions. If left untreated recurrent infections may occur, along with impaired gas exchange and in the long run fibrosis of the affected area1.A female patient at the age of 3 years with Tay-Sachs disease and consequent severe global psychomotor delay and epilepsy was initially admitted due to a respiratory infection to the neuropediatric department, where she developed an epileptic status and desaturation, prompting a transfer to the Intensive Care Unit. The refractory epileptic status resolved only with several antiepileptics, which lead to intubation requiring invasive mechanical ventilation, after which the patient stabilized.An attempt to reduce sedation the next day resulted in repeated seizures. On the third day, inflammatory parameters increased, with a radiological finding suggestive of pneumonia and a large left atelectasis confirmed by ultrasound. Antibiotics were empirically started, and Moraxella catarrhalis and Adenovirus in the respiratory tract were obtained in the cultures.Lung ultrasound showed a large consolidation in the dorsal upper two thirds of the left hemithorax. Due to the size of the atelectasis, we decided to perform bedside bronchoscopy.A single use paediatric flexible bronchoscope (Ambu® aScope™ 5 Broncho) was introduced through the ETT. The tracheobronchial morphology did not show any obstructions. We applied our recently published modified segmental insufflation technique, however without the use of surfactant2.First, a bronchoalveolar lavage was performed for microbiological and cytological analysis, followed by ultrasound guided bronchoscopic treatment of the affected area. The patient was positioned on the right lateral decubitus position. As described previously, a cuff pressure manometer (AG Cuffill Cuff) and a 50 ml syringe were attached to a three-way stopcock connected to the bronchoscope’s working channel. Pressure controlled segmental insufflation of the lingula and upper lobe was performed directly observing an aeration of the atelectatic areas on the ultrasound with no complications.Subsequent ultrasound monitoring resulted in total expansion of the affected lung and resolution of inflammation, confirmed radiologically (Figure 1.). The patient could be safely extubated and transferred to the wards. No reoccurrence of atelectasis was observed.ConclusionThe treatment of pediatric atelectasis poses a challenge, with limited approaches available and a lack of evidence-based guidelines to guide clinical decision-making. A question that remains to be answered is not only the mode of treatment, but timing. Even with bronchoscopic treatment, different methods exist. In obstructive atelectasis, the approach is simple - to eliminate the obstruction, however, nonobstructive atelectasis remain challenging. Bronchoscopic lung insufflation technique has been explored in adults, with limited cases reported in children3. Several case reports utilized pressure-controlled insufflation with or without surfactant in long standing atelectasis as an add on treatment2; 4.We believe that patients with severe neurological diseases require special attention regarding their susceptibility for developing atelectasis5. Our approach is early bronchoscopic pressure-controlled insufflation in non-obstructive atelectasis. Depending on the duration, we additionally instill surfactant in long standing ones.Ultrasound is recently being used in lung recruitment maneuvers to monitor lung expansion, however, it was only recently reported to be utilized for segmental insufflation monitoring in a spinal muscular atrophy type I patient4. However, to our best knowledge, our case report is the first successful use on a Tay-Sachs patient using real time ultrasound monitoring during pressure-controlled segmental bronchoscopic insufflation. The benefits of ultrasound guidance is the direct visualisation of lung expansion reducing the need for fluoroscopic radiation exposure of visualisation and further monitoring. Besides it increases the safety of the procedure allowing for detection of development of pneumothorax.In conclusion, pressure controlled real-time ultrasound guided bronchoscopic segmental insufflation, with its low-cost, lightweight, and flexible setup, seems to be a safe and promising new modality for treatment of non-obstructive atelectasis, especially in children requiring complex care.FIGURESFigure 1. On the left a large left sided atelectasis can be seen causing the mediastinal shift. On the right complete regression of the atelectasis is observed before extubatingReferences1. Ogake S, Bellinger C. 2020. Role of bronchoscopy in atelectasis. Clinical Pulmonary Medicine. 27(1):30-32.2. Šapina M, Olujic B, Nađ T, Vinkovic H, Dupan ZK, Bartulovic I, Milas K, Kos M, Divkovic D, Zubčić Z. 2023. Bronchoscopic treatment of pediatric atelectasis: A modified segmental insufflation‐surfactant instillation technique. Pediatric Pulmonology.3. Abu‐Hasan MN, Chesrown SE, Jantz MA. 2013. Successful use of bronchoscopic lung insufflation to treat left lung atelectasis. Pediatric Pulmonology. 48(3):306-309.4. Esteban IA, Osona B, Ballestín AS, Rubio CC, de Mendiola JMFP, Gómez VA, López ME, Vicente JCDC. 2024. Bronchoscopic segmental alveolar recruitment in a patient with spinal muscular atrophy and massive atelectasis. Pediatric pulmonology.5. St‐Laurent A, Zysman‐Colman Z, Zielinski D. 2022. Respiratory prehabilitation in pediatric anesthesia in children with muscular and neurologic disease. Pediatric Anesthesia. 32(2):228-236.

In this article we present a new method called PSF-Radon transform algorithm. This algorithm consists on recovering the instrument point spread function (PSF) from the Radon tranform (in the line direction axis) of the Line Spread function (that is, the image of a line). We present the method and tested with synthetic images, and real images from macro lens camera and microscopy. A stand-alone program along with a tutorial is available, for any interested user, in [? ] .

Aim: The 4th Davos Declaration, convened during the Global Allergy Forum (GAF) in Davos, aimed to elevate patient care for patients with atopic dermatitis (AD) by uniting experts and stakeholders. The forum addressed the high prevalence of AD, with a strategic focus on advancing research, treatment, and management to meet the evolving challenges in the field. Methods: This multidisciplinary forum brought together top leaders from research, clinical practice, policy, and patient advocacy to discuss the critical aspects of AD, including neuroimmunology, environmental factors, comorbidities, and breakthroughs in prevention, diagnosis, and treatment. The discussions were geared towards fostering a collaborative approach to integrate these advancements into practical, patient-centric care. Results The forum underlined the mounting burden of AD, attributing it to significant environmental and lifestyle changes. It acknowledged the progress in understanding AD and in developing targeted therapies but recognized a gap in translating these innovations into clinical practice. Emphasis was placed on the need for enhanced awareness, education, and stakeholder engagement to address this gap effectively and to consider environmental and lifestyle factors in a comprehensive disease management strategy. Conclusion: The 4th Davos Declaration marks a significant milestone in the journey to improve care for people with AD. By promoting a holistic approach that combines research, education, and clinical application, the Forum sets a roadmap for stakeholders to work together to improve patient outcomes in AD, reflecting a commitment to adapt and respond to the dynamic challenges of AD in a changing world.

Connectivity and data exchange are key features of Industry 4.0. In this paradigm, (Industrial) Internet of Things ((I)IoT) devices are a vital component facilitating the collection and transmission of environmental data from the physical sys- tem to the central station for processing and analysis(Digital Twin). However, although (I)IoT devices play a critical role in this process, they are not inherently equipped to run strong encryption mechanisms to secure the data they transmit over wired or wireless channels. This research aims to explore the potential of Digital Twins (DT) in securing Industry 4.0 appli- cations and the security mechanism employed to ensure con- fidentiality, integrity, and authentication of data communi- cated between (I)IoT and DT through a Systematic Literature Review (SLR). This systematic literature review, based on the analysis of 67 papers published between 2018 and 2023, un- derscores the evolving significance of Digital Twin technol- ogy, particularly within the ambit of Industry 4.0. The find- ings illuminate the pervasive influence of Digital Twin tech- nology across multiple industrial sectors. The result SLR re- vealed that DT is growing and being widely adopted as a se- curity tool particularly in Industry 4.0 using enabling tech- nology like machine learning, data analytics, blockchain, and 5G networks to provide security solutions such as intrusion detection, vulnerability assessment, cyber range, and threat intelligence.

Fish ear bones, known as otoliths, are often collected by fisheries to assist in management, and are a common sample type in museum and national archives. Beyond their utility for aging, morphological and trace element analysis, otoliths are a repository of valuable genomic information. Previous work has shown that DNA can be extracted from the trace quantities of tissue remaining on the surface of otoliths, despite the fact that they are often stored dry at room temperature. However, much of this work has used reduced-representation sequencing methods in clean lab conditions, to achieve adequate yields of DNA, libraries, and ultimately single-nucleotide polymorphisms (SNPs). Here, we pioneer the use of small-scale (spike-in) sequencing to screen otolith samples prepared in regular molecular biology laboratories for contamination and quality levels, submitting for whole-genome resequencing only samples above a defined endogenous DNA threshold. Despite the typically low quality and quantity of DNA extracted from otoliths, we are able to produce whole-genome libraries and ultimately sets of filtered, unlinked and even adaptive SNPs of ample size for downstream uses in population, climate, and conservation genomics. By comparing with a set of tissue samples from the same species, we are able to highlight the quality and efficacy of otolith DNA samples from DNA extraction and library preparation, to bioinformatic preprocessing and SNP calling. We provide detailed schematics, protocols, and scripts of our approach, such that it can be adopted widely by the community, improving the use of otoliths as a source of valuable genomic data.

Translation of messenger RNA (mRNA) in bacteria occurs in the steps of initiation, elongation, termination and ribosome recycling. The initiation step comprises multiple stages and uses a special transfer RNA (tRNA) called initiator tRNA (i-tRNA), which is first aminoacylated and then formylated using methionine and N 10-formyl-tetrahydrofolate (N 10-fTHF), respectively. Both methionine and N 10-fTHF are produced via one-carbon metabolism, linking translation initiation with active cellular metabolism. The fidelity of i-tRNA binding to the ribosomal peptidyl-site (P-site) is attributed to the structural features in its acceptor stem, and the highly conserved three consecutive G-C base pairs (3GC pairs) in the anticodon stem. While the acceptor stem region is important in formylation of the amino acid attached to i-tRNA and its initial binding to the P-site, the 3GC pairs are crucial in transiting i-tRNA through various stages of initiation. We utilized the feature of 3GC pairs to investigate the nuanced layers of scrutiny that ensure fidelity of translation initiation through i-tRNA abundance and its interactions with the components of the translation apparatus. We discuss the importance of i-tRNA in the final stages of ribosome maturation, as also the roles of the Shine-Dalgarno sequence, ribosome heterogeneity, initiation factors, ribosome recycling factor and coevolution of the translation apparatus in orchestrating a delicate balance between the fidelity of initiation and/or its leakiness to generate proteome plasticity in cells to confer growth fitness advantages in response to the dynamic nutritional states.