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We report a case of a male in his 50 s who underwent pancreaticoduodenectomy for solid pseudopapillary neoplasm (SPN) of the pancreas at 30 years. He developed a liver abscess 15 years after the surgery, and CT scan revealed a swollen retroperitoneum lymph node and a tumor in the liver. Symptoms, including abdominal distension, appetite loss, and epigastric pain, appeared due to lymph node metastasis. Endoscopic ultrasonography-guided fine-needle aspiration against the lymph node revealed SPN recurrence. The tumor had invaded the common hepatic artery, and surgery was not indicated. Chemotherapy of Gemcitabine/nab-Paclitaxel biweekly was performed 8 times; however, no reduction in tumor size was observed, and the patient's symptoms worsened. Proton beam therapy (67.5 GyE in 25 fractions) was subsequently performed for lymph node metastasis, and led to a gradual reduction in lymph node metastasis, and an improvement in symptoms. No re-expansion of lymph node metastasis has been observed 3 years after proton beam therapy. Since SPN is low malignancy and most cases can be expected to be cured by surgery, there is currently no standard treatment of unresectable SPN. This case is the first report of proton beam therapy for SPN, and was considered to be effective.

Unconventional mRNA splicing on the endoplasmic reticulum (ER) membrane is the sole conserved mechanism in eukaryotes to transmit information regarding misfolded protein accumulation to the nucleus to activate the stress response. In metazoans, the unspliced form of X-box-binding protein 1 (XBP1u) mRNA is recruited to membranes as a ribosome nascent chain (RNC) complex for efficient splicing. We previously reported that both hydrophobic (HR2) and translational pausing regions of XBP1u are important for the recruitment of its own mRNA to membranes. However, its precise location and the molecular mechanism of translocation are unclear. We show that XBP1u-RNC is specifically recruited to the ER membrane in an HR2- and translational pausing-dependent manner by immunostaining, fluorescent recovery after photobleaching, and biochemical analyses. Notably, translational pausing during XBP1u synthesis is indispensable for the recognition of HR2 by the signal recognition particle (SRP), resulting in efficient ER-specific targeting of the complex, similar to secretory protein targeting to the ER. On the ER, the XBP1u nascent chain is transferred from the SRP to the translocon; however, it cannot pass through the translocon or insert into the membrane. Therefore, our results support a noncanonical mechanism by which mRNA substrates are recruited to the ER for unconventional splicing.

Primary granulocytic sarcoma (GS) is a rare disease defined by the absence of antecedent or concomitant leukemic cells in the bone marrow and the peripheral blood. Immunohistochemical staining for myeloperoxidase is necessary for a definite diagnosis. Otherwise, primary GS is often misdiagnosed as a malignant lymphoma or other malignancies. Primary GS is well known to frequently develop into acute myeloid leukemia (AML). Here we describe a 28-year-old woman with primary GS manifesting as an epidural tumor in the sacral region accompanied by meningeal dissemination. Fluorescence in situ hybridization analysis detected the AML1/MTG8 fusion gene in neoplastic cells obtained from her cerebrospinal fluid specimen and the epidural mass. The AML1/MTG8 fusion gene transcript was also detected by a nested reverse transcriptase-polymerase chain reaction analysis of mononuclear cells from the bone marrow, although leukemic cells were not recognized in a microscopical examination of the patient's bone marrow. Systemic chemotherapy with high-dose cytarabine followed by local radiotherapy was performed, and the patient clinically achieved a complete response. These molecular analyses provide a precise method of diagnosis, especially with respect to the French-American-British AML classification, according to the characteristic karyotypic alterations, and a patient consequently can quickly be given appropriate systemic chemotherapy as induction therapy.

The frequency of t(14;18) in follicular lymphoma (FL) in Japan has been reported to be low compared to North America and other European countries. Recently, it has also been reported that FL lacks t(14;18), mainly among histological grade 3b, and occasionally has a rearranged Bcl-6 gene. It is not known whether a difference in histology or immunostaining pattern exists between FL with and without t(14;18). We performed interphase fluorescence in situ hybridization (FISH) analysis to detect Bcl-2/IgH, Bcl-6 gene rearrangement, Bcl-2 gene amplification, and the cyclinD1/IgH gene in formalin-fixed paraffin embedded specimens from our FL archives. The correlation between morphological features, histological grades, immunohistochemical findings, and cytogenetical aberrations was studied. In total, we found that 28 of 47 cases (59.6%) had t(14;18). Bcl-6 gene rearrangement and extra Bcl-2 gene signals were found in five and two cases, respectively. Only one had cyclinD1/IgH fusion. Ten of 12 grade 1, nine of 17 grade 2, and 0 of two grade 3 cases had fusion signals, respectively. None of the above abnormalities were detected in 12 of 47 cases (25.5%). Our data confirmed a high frequency of t(14;18) in FL in grade 1, but a lower incidence among grade 2, that could be attributed to the lower incidence of the translocation in FL in Japan. Immunostaining of both Bcl-2 and CD10 was highly predictable for the presence of t(14;18); the positive predictive value was 75%, suggesting the usefulness of the staining.

Mucosa-associated lymphoid tissue (MALT) lymphoma is a low-grade B-cell lymphoma and is usually slow to disseminate. The t(11;18)(q21;q21) translocation has been identified in a subset of MALT lymphoma cases, and AP12 and MLT1/MALT1 genes have been implicated in this translocation. However, the clinicopathologic features of t(11;18)-bearing MALT lymphoma have not been fully elucidated, and the optimal therapy for patients with disseminating disease is unknown. We report an outstanding case of MALT lymphoma that showed massive pulmonary infiltration and leukemic transformation during a prolonged course of more than 16 years. Reverse transcriptase-polymerase chain reaction and dual-color fluorescence in situ hybridization analyses confirmed the existence of the AP12/MLT1 fusion gene in the lymphoma cells. Peripheral blood lymphoma cells disappeared after glucocorticoid therapy. Although the pulmonary lymphoma was slowly progressive and caused severe hypoxemia despite the continuation of glucocorticoid therapy, treatment with cladribine induced a marked reduction of pulmonary lymphomatous lesions and an improvement of the hypoxemia. These findings show the unique clinical features of this particular indolent B-cell lymphoma with t(11;18) translocation and suggest the potential therapeutic usefulness of glucocorticoid and cladribine.

In order to create an ochre suppressor tRNA which exclusively recognizes UAA codon, we replaced the G34 at the first position of yeast tRNA(Tyr)[GPsiA] anticodon with pseudouridine34 (Psi34) by using the molecular surgery technique. This tRNA(Tyr)[PsiPsiA] recognized only the UAA codon as expectedly, but tRNA(Tyr)[UPsiA] made as a control also behaved similarly. This result may suggest that U34 must be somehow modified to facilitate the wobble-pairing to G at the third position of codon.

Background: Recurrent karyotypic aberrations are shown in MALT-type lymphomas. It is unknown whether these heterogeneities are translated into difference of expression profiles creating distinct subtypes. Ocular adnexal MALT-type lymphoma (OAL) lacks contaminated extra-organ tissue, providing an opportunity to perform array analysis of pure tumor cell component. We have previously reported OAL cases with trisomy 18 might constitute a unique clinicopathological entity with shortened progression-free survival (Blood2005; 106: 289a). Patients and Methods: Archives of samples of OAL of our institute between 1995 and 2003 were utilized. Karyotypic informations were obtained by interphase fluorescence in situ hybridization (FISH) analysis. Oligonucleotide microarray with U133 Plus 2.0 Array (Affymetrix) were carried out. Statistical analysis was performed using GeneSpring GX 7.3.1 software. Biological networks were generated with Ingenuity Pathway Analysis version 3.1. Results: A total of 26 cases of OAL were analyzed. Eighteen were male and 8 were female, with an overall median age of 56 years at the time of diagnosis (range, 30–90 years). Clinical stage was I (23 cases), II (1 case), and IV (2 cases). There were 1 case of IgH/MALT1 fusion, 16 cases of trisomy 3, 11 cases of trisomy 18, and 2 cases of trisomy 7 by FISH analysis. No cases had API2/MALT1 fusion or trisomy 12. In an unsupervised method, no clusters were identified within the categories including sex, age, anatomical site, trisomy3, 18, pathological features and progression. In a supervised method, a set of differentially expressed genes were identified which accurately stratified samples categories of trisomy 18 and others. Hierarchical clustering and principal component analysis of 26 OAL samples based on the differential expression of the 2506 genes, the two distinct subclasses accurately clustered together. The percentage of samples correctly predicted was 92% with leave-one-out cross validation using support vector machine, further confirming that our prior clustering very accurately predicted class assignment of OAL with trisomy 18 and that without trisomy 18 subset. The top three genes with high expression were HLA-DRB3, unidentified gene KIAA0924, and apoptosis related gene, MDM4. A network analysis was performed to identify differently expressing genes related to apoptosis and proliferation. A case with t(14;18) showed over-expression of MALT1 gene and CARD11, as might be expected. Trisomy 18 cases had higher expression of Bcl-2 gene comparing to others. Other genes with higher expression shown in trisomy 18 in this network were anti-apoptotic genes including IKBKbeta and XIAP. A down-regulated gene was RTN4, functioning as down regulator of Bcl-2 gene. Cell cycle regulating genes including cyclin D2, CDK2, CDK6 and cyclin A2 were also highly expressed in OAL with trisomy18. Conclusions: Our expression profile analysis again revealed uniqueness of trisomy 18 OAL. This might be attributed to the different levels of anti-apoptosis and proliferation gene-expressions.

Background: It remains unknown whether primary ocular adnexal MALT lymphoma is a homogenous disease, because there have been few reports on cytogenetic or molecular analysis of this disease. Patients and Methods: In 34 cases of ocular adnexal MALT lymphoma, we performed interphase fluorescence in situ hybridization (FISH) analysis to detect IgH/MALT1 and API2/MALT1 fusion genes. Aneuploidy of chromosome 3, 7, 12 and 18 was also identified using corresponding centromere probes. We defined trisomy when three centromeres were recognized in FISH analysis. Histopathologic features were reviewed and categorized according to the degree of plasma cell differentiation, monocytoid B cell feature, nodularity, abundance of reactive germinal centers, multikaryocytic cells, contaminated T cells, Duthcer bodies, and lymphoepithelial lesions (LELs). Correlations among FISH analysis, histopathologic features, and clinical data were analyzed. Results: FISH analysis showed 1 (3%) IgH/MALT1 fusion, 21 (62%) trisomy 3, 16 (47%) trisomy 18, and 3(9%) trisomy 7 cases. No cases showed API2/MALT1 fusion or trisomy 12. Existence of these cytogenetic change did not influence the degree of morphological features significantly, except for trisomy 18; Cases with trisomy 18 had significantly more abundant LEL cells, monocytoid B cells and residual reactive germinal centers, but less nodules, contaminated plasma cells and large cells. Cases with trisomy 18 showed distinct features of female dominance, younger age, and included more cases originated from conjunctiva. In total, five (15%) of 34 patients relapsed between 21 and 103 months, and all of them were found to have trisomy 18. Conclusions: Aneuploidy is found in a ceratin subset of ocular adnexal MALT lymphoma by FISH analysis. Cases with trisomy18 may make a distinct clinicopathlogic entity.

Introduction: Amplification and translocation of bcl-2 gene have been detected in a certain subset of diffuse large B-cell lymphoma (DLBCL). Recently, it has been reported that tissue microarray might be able to divide DLBCL into significant subgroups similar to those determined by cDNA array. The correlations among bcl-2 protein expression, gene translocation or amplification, and molecular signature determined by cDNA array are poorly understood. Materials and methods: This study included 25 cases with de novo nodal DLBCL which were diagnosed between June 1997 and December 2001, and had been followed up at a single institution (NCCH). There were 12 males and 13 females with a median age at diagnosis of 60 years (range, 24 to 83 years). All the tissue samples were formalin-fixed paraffin sections obtained by biopsy before chemotherapy. Interphase fluorescence in situ hybridization (FISH) analysis were performed to evaluate the bcl-2 gene status using IGH/BCL2 and CEP 18 centromere probe (Vysis). We named a “additional chromosome 18” when extra BCL2 gene signals were observed in each tumor cells in proportion to extra signals of CEP18 probe, and “gene amplification” when extra BCL2 signals were observed without additional CEP18 signals. To define each case as “Germinal Center B-cell (GCB) phenotype” or “non-GCB phenotype” a panel of 3 antigens, CD10, bcl-6, and MUM-1 was evaluated following the algorithm reported by Hans CP et al (Blood.2004;103:275). Immunoreactivity was judged to be positive if 20% or more of the tumor cells were stained, and strongly positive if 50% or more. Results: CD10, bcl-6, and MUM1 were positive in 5/25 (20%), 25/25 (100%), and 17/25 (68%) cases, respectively. Nineteen of 25 (76%) cases were positive, 13 of 25 (52%) strongly positive for the bcl-2 protein staining. Of 25 cases in total, 8 cases (32%) were classified into “GCB phenotype” and 17 cases (68%) were classified into “non-GCB phenotype”. FISH analysis revealed that t(14;18) was detected in 2 of 8 cases (25%) of “GCB phenotype”, but in none of 17 cases of “non-GCB phenotype”. Extra BCL2 gene signals were detected in 7 of 25 (28%); “additional chromosome 18” (n=5), “gene amplification” (n=1), and “additional chromosome 18 + gene amplification” (n=1). “Additional chromosome 18” and “gene amplification” were exclusively detected in “non-GCB phenotype” DLBCL, and these cases showed strongly positive bcl-2 staining except for one. Thirteen cases with strongly positive bcl-2 staining included t(14;18) (n=2) and extra signals of BCL2 (n=6). No cases showed both t(14;18) and extra BCL2 gene signals, simultaneously. Conclusion: Our results showed t(14;18) is found among cases with “GCB phenotype”, whereas, additional chromosome 18 and/or bcl-2 gene amplification, among “non-GCB phenotype”. The mechanism for overexpression of bcl-2 protein in de novo DLBCL varies between “GCB phenotype” and “non-GCB phenotype” defined by immunohistochemistry.

Background It remains unknown whether primary ocular adnexal extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) is a homogeneous entity, as there are few reports of the results of cytogenetic or molecular analyses of these tumors. Methods We performed interphase fluorescence in situ hybridization (FISH) analysis to detect translocations and aneuploidy in 34 cases of primary ocular adnexal MALT lymphoma, and reviewed the histopathological findings. Correlations between the results of FISH analysis, the histopathological features and the clinical data were also analyzed. Results Among the 34 cases, FISH analysis revealed t(14;18)(q32;q21) in one case, trisomy 3 in 21 cases (62%), and trisomy 18 in 16 cases (47%). The cases with trisomy 18 had significantly more prominent lymphoepithelial lesions (LELs) and less nodularity in the tumors. In regard to the clinical correlations, tumors with trisomy 18 were observed predominantly in females and younger patients; also, in the majority of the cases, the tumor was of conjunctival origin. All the cases with recurrence showed trisomy 18 in the tumor. Conclusion Primary ocular adnexal MALT lymphoma is a significantly heterogeneous entity. Cases with trisomy 18 may have unique clinicopathological features.

Amplification and translocation of the Bcl-2 gene has been detected in a certain subset of diffuse large B-cell lymphomas (DLBCL). The correlations among Bcl-2 protein expression, gene translocation or amplification, and the molecular signature determined by cDNA array are poorly understood. This study examined 25 cases with de novo nodal DLBCL. Interphase fluorescence in situ hybridization (FISH) analysis was performed to evaluate the Bcl-2 gene using IGH/BCL2 and CEP18 centromere probes (Vysis). When extra Bcl-2 gene signals were observed in each tumor cell and when these signals were in proportion to the extra CEP18 probe signals, we regarded the findings as indicating the presence of an additional chromosome 18; when extra Bcl-2 signals were observed but additional CEP18 signals were not, we regarded the findings as indicating the presence of gene amplification. A panel of 3 antigens (CD10, Bcl-6, and MUM-1) was applied to categorize each case as either a "germinal center B-cell (GCB) phenotype" or a "non-GCB phenotype." Of the 25 cases examined, 8 cases (32%) were classified as "GCB phenotype" and 17 cases (68%) were classified as "non-GCB phenotype." A FISH analysis revealed that t(14;18) was detected in 2 of the 8 cases (25%) with the "GCB phenotype" but in none of the 17 "non-GCB phenotype" cases. Extra Bcl-2 gene signals were detected in 7 of the 25 (28%) cases examined: n = 5 for an additional chromosome 18, n = 1 for gene amplification, and n = 1 for additional chromosome 18 + gene amplification. Extra Bcl-2 gene signals were exclusively detected in DLBCL with the "non-GCB phenotype"; these cases, with the exception of one, stained strongly positive for Bcl-2. The DLBCLs with Bcl-2 protein overexpression were classified into at least two heterogeneous molecular groups, based on the results of the FISH analysis.