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Objective: To investigate the clinicopathological features and differential diagnosis of eosinophilic vacuolated tumor (EVT). Methods: Seven cases of EVT with characteristic morphology and unequivocal diagnosis from the Affiliated Hospital of Qingdao University (6 cases), Qingdao, China and the 971 Hospital of PLA Navy (1 case), Qingdao, China between January 2010 and December 2021 were subject to morphological and immunohistochemical analyses. Additionally, whole exome sequencing (WES) was performed in two cases. Twenty-two cases of renal oncocytoma (RO) and 17 cases of eosinophilic chromophobe renal cell carcinoma (eChRCC) diagnosed at the same time were used as controls. Results: Four males and three females with a mean age of 42 years (range: 29-61 years) were included in the study. The tumors were nodular and well-circumscribed, with sizes ranging from 1.5 to 4.5 cm. On cross-section, they appeared gray-red or gray-white, solid, and soft. Tumor cells were arranged in nests, solid sheets, and acinar or small vesicular structures. These cells exhibited eosinophilic cytoplasm with large, prominent clear vacuoles and round nuclei with prominent nucleoli. Perinuclear halos were focally present in four cases, while small tumor cells with sparse cytoplasm and hyperchromatic nuclei were seen in one case. No necrosis or mitosis was noted. Edematous stroma was detected in three cases. All tumors were positive for CD117 and Cathepsin K, but negative for vimentin and CK7. CK20 was positive in scattered individual cells, and Ki-67 positivity ranged from 1% to 4%. Point mutations in MTOR were identified in both patients who were subject to the molecular analysis. Statistical differences in the expression of Cathepsin K, CD10, S-100A1, and Cyclin D1 between EVT and RO (P<0.05) were significant, so were the differences in the expression of Cathepsin K, CD10, CK7 and claudin 7 between EVT and eChRCC (P<0.001). Seven patients were followed up for 4 to 96 months (mean, 50 months), with no recurrences or metastases. Conclusions: EVT is a rare renal tumor that shares morphological and immunophenotypic features with RO and eChRCC, and it is closely linked to the TSC/MTOR pathway. The presence of large prominent transparent vacuoles in eosinophilic cytoplasm along with conspicuous nucleoli is its key morphological characteristics. The use of combined immunohistochemical stains greatly aids in its diagnosis. Typically, the tumor exhibits indolent biological behaviors with a favorable prognosis.
目的： 探讨肾嗜酸性空泡状肿瘤（eosinophilic vacuolated tumor，EVT）的临床病理学特点及诊断、鉴别诊断。 方法： 收集2010年1月至2021年12月青岛大学附属医院（6例）和海军第九七一医院（1例）形态学典型、病理诊断无异议的肾EVT，进行组织形态学观察、免疫组织化学染色，并对2例进行全外显子测序。另选取同期诊断的肾嗜酸细胞瘤（RO）22例、嗜酸型嫌色细胞肾细胞癌（eChRCC）17例进行比较。 结果： 男性4例，女性3例。年龄29~61岁（平均年龄42岁）。临床上，7例均系体检发现，瘤体直径1.5~4.5 cm，结节状，边界清楚，切面灰红、灰白色，实性，质软。镜下观察：瘤组织呈巢团状或实性排列，伴有多少不等的腺泡状或小囊状结构，瘤细胞胞质嗜酸，胞质内均可见大而突出的透明空泡，核圆形，核仁明显，4例局部可见核周空晕，1例见灶状分布、胞质稀少、核深染的小细胞成分。所有病例均未见坏死和核分裂象。3例存在水肿性间质。免疫组织化学染色：7例肿瘤均呈CD117、Cathepsin K阳性，波形蛋白、细胞角蛋白（CK）7阴性，CK20散在或斑片状阳性，Ki-67阳性指数1%~4%。全外显子测序显示2例肿瘤均存在MTOR基因突变。统计学分析表明，EVT与RO中Cathepsin K、CD10、S-100A1和Cyclin D1的表达（P<0.05）及EVT与eChRCC中Cathepsin K、CD10、CK7和claudin7的表达（P<0.001）差异有统计学意义。7例随访4~96个月（平均50个月），均未见复发或转移。 结论： 肾EVT是一种在形态学、免疫表型与RO、eChRCC均有重叠、与TSC/MTOR通路密切相关的少见肾肿瘤，嗜酸性胞质内可见大而突出的透明空泡以及存在显著的核仁是该肿瘤的重要组织学特征。联合应用系列免疫组织化学抗体有助于EVT的诊断与鉴别诊断。该肿瘤生物学行为通常惰性，预后良好。.

Ubiquitination is a posttranslational modification in eukaryotes that plays a significant role in the infection of intracellular microbial pathogens, such as Legionella pneumophila. While the Legionella-containing vacuole (LCV) is coated with ubiquitin (Ub), it avoids recognition by autophagy adaptors. Here, we report that the Sdc and Sde families of effectors work together to build ubiquitinated species around the LCV. The Sdc effectors catalyze canonical polyubiquitination directly on host targets or on phosphoribosyl-Ub conjugated to host targets by Sde. Remarkably, Ub moieties within poly-Ub chains are either modified with a phosphoribosyl group by PDE domain-containing effectors or covalently attached to other host substrates via Sde-mediated phosphoribosyl-ubiquitination. Furthermore, these modifications prevent the recognition by Ub adaptors and therefore exclude host autophagy adaptors from the LCV. In this work, we shed light on the nature of the poly-ubiquitinated species present at the surface of the LCV and provide a molecular mechanism for the avoidance of autophagy adaptors by the Ub-decorated LCV.

The Legionella pneumophila Sde family of translocated proteins promotes host tubular endoplasmic reticulum (ER) rearrangements that are tightly linked to phosphoribosyl-ubiquitin (pR-Ub) modification of Reticulon 4 (Rtn4). Sde proteins have two additional activities of unclear relevance to the infection process: K63 linkage-specific deubiquitination and phosphoribosyl modification of polyubiquitin (pR-Ub). We show here that the deubiquitination activity (DUB) stimulates ER rearrangements while pR-Ub protects the replication vacuole from cytosolic surveillance by autophagy. Loss of DUB activity is tightly linked to lowered pR-Ub modification of Rtn4, consistent with the DUB activity fueling the production of pR-Ub-Rtn4. In parallel, phosphoribosyl modification of polyUb, in a region of the protein known as the isoleucine patch, prevents binding by the autophagy adapter p62. An inability of Sde mutants to modify polyUb results in immediate p62 association, a critical precursor to autophagic attack. The ability of Sde WT to block p62 association decays quickly after bacterial infection, as predicted by the presence of previously characterized L. pneumophila effectors that inactivate Sde and remove polyUb. In sum, these results show that the accessory Sde activities act to stimulate ER rearrangements and protect from host innate immune sensing in a temporal fashion.

An 83-year-old man with a history of monoclonal gammopathy of unknown significance, macrocytic anemia, and cytopenias presented with hemorrhagic bullae on his left hand, alongside intermittent fevers and joint pain. Laboratory findings indicated anemia, elevated mean corpuscular volume, thrombocytopenia, leukopenia, and a high C-reactive protein level. A biopsy of the bullae showed neutrophilic dermatosis, and computed tomography scans of the thorax revealed lung opacities and mediastinal lymphadenopathy, suggesting neutrophilic alveolitis. Bone marrow examination found hypercellularity with myeloid and histiocytic hyperplasia, vacuolated precursors, and 3% blasts of an immature myelomonocytic lineage. Genetic testing uncovered a UBA1 mutation at an 81% allele frequency, confirming a diagnosis of VEXAS syndrome. Treatment commenced with prednisone, initiated at 60 mg daily and tapered to 10 mg, with tocilizumab considered for future symptom management. This regimen has successfully maintained remission, as observed in follow-up appointments. This case highlights the diagnostic complexity and effective management of VEXAS syndrome, underscoring the importance of genetic testing in guiding treatment decisions.
VEXAS syndrome, initially identified in 2020, is a rare disease characterized by overlapping hematologic and rheumatologic conditions, primarily affecting middle-aged to older men.Our case presents a unique instance of VEXAS syndrome, featuring inflammatory symptoms alongside pancytopenia and a history of monoclonal gammopathy of unknown significance, highlighting the diverse clinical spectrum of this condition.The diagnosis was confirmed through bone marrow biopsy revealing vacuoles in myeloid precursors and a UBA1 mutation, reaffirming the diagnostic criteria outlined for VEXAS syndrome.Management challenges persist, with current treatment options limited to glucocorticoids for symptomatic relief, underscoring the need for further research into targeted therapies such as ruxolitinib to improve patient outcomes.

We previously reported autophagy-mediated degradation of nuclei, nucleophagy, in the filamentous fungus Aspergillus oryzae. In this study, we examined whether nuclei are degraded as a whole. We generated A. oryzae mutants deleted for orthologs of Saccharomyces cerevisiae YPT7 and ATG15 which are required, respectively, for autophagosome-vacuole fusion and vacuolar degradation of autophagic bodies. Degradation of histone H2B-EGFP under starvation conditions was greatly decreased in the ΔAoypt7 and ΔAoatg15 mutants. Fluorescence and electron microscopic observations showed that autophagosomes and autophagic bodies surrounding the entire nuclei were accumulated in the cytoplasm of ΔAoypt7 and the vacuole of ΔAoatg15, respectively. These results indicate that nuclei are engulfed in the autophagosomes as a whole and transported/released into the vacuolar lumen where they are degraded.

Plants have developed precise defense mechanisms against cadmium (Cd) stress, with vacuolar compartmentalization of Cd2+ being a crucial process in Cd detoxification. The transport of Cd into vacuoles by these cation / H+ antiporters is powered by the pH gradient created by proton pumps. In this study, the full-length cDNA of a vacuolar H+-pyrophosphatase (V-PPase) gene from Boehmeria nivea (ramie), BnVP1, was isolated using the rapid amplification of cDNA ends (RACE) method. The open reading frame (ORF) of BnVP1 is 2292 bp, encoding a 763 amino acid V-PPase protein with 15 predicted transmembrane domains. Sequence alignment and phylogenetic analysis revealed that BnVP1 belongs to the Type I V-PPase family. Quantitative RT-PCR assays demonstrated that BnVP1 expression was significantly higher in ramie roots than in shoots. Cd treatments markedly induced BnVP1 expression in both roots and leaves of ramie seedlings, with a more pronounced effect in roots. Additionally, BnVP1 expression was significantly upregulated by the plant hormone methyl jasmonate (MeJA). Heterologous expression of BnVP1 in transgenic Arabidopsis significantly enhanced V-PPase activity in the roots. The growth performance, root elongation, and total chlorophyll content of transgenic plants with high tonoplast H+-PPase (V-PPase) activity were superior to those of wild-type plants. Overexpression of BnVP1 reduced membrane lipid peroxidation and ion leakage, and significantly increased Cd accumulation in the roots of transgenic Arabidopsis seedlings. This study provides new genetic resources for the phytoremediation of Cd-contaminated farmland.

Coxiella burnetii is a globally distributed obligate intracellular pathogen. Although often asymptomatic, infections can cause acute Q fever with influenza-like symptoms and/or severe chronic Q fever. Coxiella burnetii develops a unique replicative niche within host cells called the Coxiella-containing vacuole (CCV), facilitated by the Dot/Icm type IV secretion system translocating a cohort of bacterial effector proteins into the host. The role of some effectors has been elucidated; however, the actions of the majority remain enigmatic and the list of true effectors is disputable. This study examined CBU2016, a unique C. burnetii protein previously designated as an effector with a role in infection. We were unable to validate CBU2016 as a translocated effector protein. Employing targeted knock-out and complemented strains, we found that the loss of CBU2016 did not cause a replication defect within Hela, THP-1, J774, or iBMDM cells or in axenic media, nor did it affect the pathogenicity of C. burnetii in the Galleria mellonella infection model. The absence of CBU2016 did, however, result in a consistent decrease in the size of CCVs in HeLa cells. These results suggest that although CBU2016 may not be a Dot/Icm effector, it is still able to influence the host environment during infection.

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The pistil is the most important organ for fertilization in flowering plants, and the stigmatic papilla cells are responsible for pollen acceptance and pollen tube germination. Arabidopsis plants possess dry stigmas exhibiting high selectivity for compatible pollen. When compatible pollens are recognized and accepted by stigmatic papilla cells, water and nutrients are then transported from the stigma to pollen grains through the secretory pathway. Here, we present light microscopy-based methods for investigating autophagy and senescence of stigmatic papilla cells. These methods include the assessment of viability of stigmatic papilla cells using dual staining with fluorescein diacetate/propidium iodide, as well as the examination of vacuolar-accumulated proteins during stigma senescence. These methods can be used to understand the functions of the stigma tissue from a subcellular perspective.