Mutational analysis of DNMT3A gene in acute leukemias and common solid cancers

DNMT3A, a DNA methyltransferase that functions for de novo methylation, is important in development and many cellular processes related to tumorigenesis. Somatic mutations of DNMT3A gene, including recurrent mutations in its Arg‐882, were recently reported in acute myelogenous leukemia (AML), strongly suggesting its role in development of AML. To see whether DNMT3A mutation occurs in other malignancies as well, we analyzed DNMT3A in 916 cancer tissues from 401 hematologic malignancies (AML, acute lymphoblastic leukemias (ALL), multiple myelomas and lymphomas) and 515 carcinomas (lung, breast, prostate, colorectal and gastric carcinomas) using a single‐strand conformation polymorphism (SSCP) assay. We identified DNMT3A mutations, especially the Arg‐882 mutations, in adulthood AML (9.4%). In addition, we found DNMT3A mutations in pre‐B‐ALL and three lung cancers at lower frequencies. Allelic loss of DNMT3A was frequently observed in most cancer types analyzed, including lymphomas (48.1%), gastric cancers (23.5%) and lung cancers (18.3%) irrespective of DNMT3A mutation. Also, loss of DNMT3A expression was common in lung cancers (46.4%), and was associated with the allelic loss. Our data indicate that DNMT3A gene is mutated mainly in AML, but it occurs in other cancers, such as ALL and lung cancer, despite the lower incidences. Also, the data suggest that DNMT3A is altered in many cancer types by various ways, including somatic mutations, allelic loss and loss of expression that might play roles in tumorigenesis.     

Cucurbita moschata Duch. and its active component, β-carotene effectively promote the immune responses through the activation of splenocytes and macrophages

Cucurbita moschata Duch. has long been used for traditional health food in many countries. However, to enhance the immune system of Cucurbita moschata Duch. and its major component, β-carotene is not clear. Here, we determined the immune enhancement effect of Cucurbita moschata Duch. and β-carotene in mouse splenocytes and RAW 264.7 macrophage cell line. We prepared baked Cucurbita moschata Duch. (Sweetme Sweet Pumpkin^TM, SSP) and steamed Cucurbita moschata Duch. (SC). Splenocytes isolated from the spleen of BALB/c mice were treated with SSP, SC, and β-carotene for 24?h. RAW 264.7 cells were stimulated with recombinant interferon-γ (rIFN-γ) for 6?h before treatment with SSP, SC, or β-carotene. SSP, SC and β-carotene significantly up-regulated the proliferation of splenocyte and mRNA expression of KI-67. The levels of interleukin-2 and IFN-γ were up-regulated by SSP, SC, or β-carotene in the splenocytes. SC and β-carotene also increased the levels of tumor necrosis factor-α (TNF-α) in the splenocytes. In addition, SSP, SC, or β-carotene significantly increased the levels of TNF-α through the nuclear translocation of the nuclear factor-κB and phosphorylation of IκBα in the rIFN-γ-primed RAW 264.7 cells. These data indicate that Cucurbita moschata Duch. and β-carotene may have an immune-enhancing effect through the production of Th1 cytokines by activation of splenocytes and macrophages.     

Differential diagnosis of jaw pain using informatics technology

This study aimed to deduce evidence‐based clinical clues that differentiate temporomandibular disorders (TMD)‐mimicking conditions from genuine TMD by text mining using natural language processing (NLP) and recursive partitioning. We compared the medical records of 29 patients diagnosed with TMD‐mimicking conditions and 290 patients diagnosed with genuine TMD. Chief complaints and medical histories were preprocessed via NLP to compare the frequency of word usage. In addition, recursive partitioning was used to deduce the optimal size of mouth opening, which could differentiate TMD‐mimicking from genuine TMD groups. The prevalence of TMD‐mimicking conditions was more evenly distributed across all age groups and showed a nearly equal gender ratio, which was significantly different from genuine TMD. TMD‐mimicking conditions were caused by inflammation, infection, hereditary disease and neoplasm. Patients with TMD‐mimicking conditions frequently used “mouth opening limitation” (P < .001), but less commonly used words such as “noise” (P < .001) and “temporomandibular joint” (P < .001) than patients with genuine TMD. A diagnostic classification tree on the basis of recursive partitioning suggested that 12.0 mm of comfortable mouth opening and 26.5 mm of maximum mouth opening were deduced as the most optimal mouth‐opening cutoff sizes. When the combined analyses were performed based on both the text mining and clinical examination data, the predictive performance of the model was 96.6% with 69.0% sensitivity and 99.3% specificity in predicting TMD‐mimicking conditions. In conclusion, this study showed that AI technology‐based methods could be applied in the field of differential diagnosis of orofacial pain disorders.     

Isolation and characterisation of microsatellite loci in the tire track eel,Mastacembelus favus and cross-species amplification

Nine microsatellite loci were isolated and characterized in the tire track eel, Mastacembelus favus by using library enriched for microsatellite region. Among 47 specimens collected from Mekong River, the number of alleles ranged from 2 to 17 while levels of observed and expected heterozygosities ranged from 0.021 to 0.873 and from 0.021 to 0.893, respectively. The primers were successfully tested in six closely related species, hence valuable for investigating population genetics and other related aspects of M. favus and its congener species.     

A Case of Paclitaxel-induced Maculopathy Treated with Methazolamide

A 54-year-old female patient who had been undergoing anti-cancer chemotherapy and radiotherapy for seven years after surgery for left breast cancer visited our clinic for visual disturbance in the right eye at nine months after paclitaxel administration. The best-corrected visual acuity was 0.5 in the right eye and 1.0 in the left eye. The patient was diagnosed with maculopathy due to paclitaxel administration based on the finding of cystoid macular edema in the right eye on fundus examination and optical coherence tomography; however, no leakage was detected on fluorescein angiography. Thus, drug replacement was planned. On the other hand, no abnormal finding was observed in the left eye. However, as the anti-cancer effect of paclitaxel is significant, replacing paclitaxel with another agent was not warranted; therefore, maintenance therapy with methazolamide was performed before and after administering the anti-cancer agent. Aggravation of cystoid macular edema was prevented, and vision improvement was achieved by oral maintenance therapy with methazolamide. In addition, the same fundus findings as shown in the right eye were detected in the left eye at 16 months after paclitaxel administration. After administering methazolamide, macular thickness was reduced, and vision was improved in the left eye. Paclitaxel administration was discontinued due to cutaneous metastasis from the breast cancer, and another anti-cancer agent was then administered. No subsequent cystoid macular edema has occurred.     

Conditional ligands for Asian HLA variants facilitate the definition of CD8+ T‐cell responses in acute and chronic viral diseases

Conditional ligands have enabled the high‐throughput production of human leukocyte antigen (HLA) libraries that present defined peptides. Immunomonitoring platforms typically concentrate on restriction elements associated with European ancestry, and such tools are scarce for Asian HLA variants. We report 30 novel irradiation‐sensitive ligands, specifically targeting South East Asian populations, which provide 93, 63, and 79% coverage for HLA‐A, ‐B, and ‐C, respectively. Unique ligands for all 16 HLA types were constructed to provide the desired soluble HLA product in sufficient yield. Peptide exchange was accomplished for all variants as demonstrated by an ELISA‐based MHC stability assay. HLA tetramers with redirected specificity could detect antigen‐specific CD8⁺ T‐cell responses against human cytomegalovirus, hepatitis B (HBV), dengue virus (DENV), and Epstein‐Barr virus (EBV) infections. The potential of this population‐centric HLA library was demonstrated with the characterization of seven novel T‐cell epitopes from severe acute respiratory syndrome coronavirus, HBV, and DENV. Posthoc analysis revealed that the majority of responses would be more readily identified by our unbiased discovery approach than through the application of state‐of‐the‐art epitope prediction. This flow cytometry‐based technology therefore holds considerable promise for monitoring clinically relevant antigen‐specific T‐cell responses in populations of distinct ethnicity.