Sphingosine-1-phosphate receptor 1 as a prognostic biomarker and therapeutic target for patients with primary testicular diffuse large B-cell lymphoma.

Sphingosine-1-phosphate (S1P) is a potent lipid mediator that is produced during the metabolism of sphingolipid by sphingosine kinase. S1P has been implicated in the migration and trafficking of lymphocytes and several lymphoid malignancies through S1P receptors. Moreover, the overexpression of sphingosine-1-phosphate receptor 1 (S1PR1) has been correlated with the constitutive activation of signal transducer and activator of transcription (STAT)3 and poor prognosis of diffuse large B-cell lymphoma (DLBCL). Thus, in this study, we examined the expression of S1PR1 in 198 DLBCL samples collected from nodal and various extranodal sites and sub-classified formalin-fixed paraffin-embedded tissue samples into germinal centre B-cell-like (GCB) and non-GCB subgroups using immunohistochemistry. These analyses showed S1PR1 overexpression in 15·7% of all cases with DLBCL and in 54·2% of 24 cases with primary testicular (PT)-DLBCL; S1PR1 expression correlated with S1PR1mRNA expression and STAT3 phosphorylation in fresh samples. Analyses of data from a single institution suggested that S1PR1 overexpression was an independent negative prognostic marker in 68 patients with DLBCL of clinical stages I and II. The present high prevalence of S1PR1 overexpression warrants the consideration of PT-DLBCL as a distinct disease subtype and suggests the potential of the S1P/S1PR1 axis as a therapeutic target.

Poikilodermatous mycosis fungoides with a CD8+ CD56+ immunophenotype: a case report and literature review.

Mycosis fungoides (MF) represents the most common type of cutaneous lymphoma. MF shows varieties in both its clinical presentation and immunophenotype. We herein report one case of poikilodermatous MF with a CD8+ CD56+ immunophenotype and present a literature review. A 20-year-old Japanese woman presented with a 10-year history of multiple poikilodermatous and reddish or brownish patches with mild pruritus on the chest, abdomen, back, buttock and thighs. Histopathologically, small- to medium-sized atypical lymphocytes infiltrated into the epidermis, indicating epidermotropism, along the basal layer, and distributed in band-like appearance in the papillary dermis. Immunohistochemically, atypical lymphocytes expressed CD3, CD8, CD56, T-cell intracellular antigen (TIA)-1, granzyme B and beta F1 but lacked expression of CD4, CD20, CD30 and Epstein-Barr virus (EBV) latent membrane protein 1. An EBV-encoded small non-polyadenylated RNA-1 (EBER-1) signal was not detected. On the basis of these findings, the diagnosis of CD8+ CD56+ MF was established. Poikilodermatous MF with a CD8+ CD56+ immunophenotype, as presented herein, is extremely rare. Although further investigation is needed to fully clarify the nature of this aberrant phenotype of MF, we stress that it is important to recognize this rare immunophenotype of MF to distinguish it from aggressive cytotoxic cutaneous lymphomas.

Relationship between gastroesophageal junction adenocarcinoma and Helicobacter pylori infection in Japan.

BACKGROUND/AIMS: The relationship between gastroesophageal junction adenocarcinoma (GEJA) and Helicobacter pylori infection is not well defined; thus, we retrospectively investigated this relationship. METHODS: We examined 852 cases (646 men) of gastric cancer. GEJA was defined as type II according to the classification system of Siewert and Stein. We compared the prevalence of H. pylori infection and corporal gastritis in GEJA patients with distal gastric cancer. RESULTS: GEJA was observed in 80 (including 6 cases of Barrett's esophageal cancer) of the 852 cases of gastric cancer examined (9.4%). The rate of H. pylori infection was significantly lower in patients with GEJA than in patients with distal gastric cancer (73.8 vs. 94.1%, p < 0.05). The prevalence of corporal gastritis was also significantly lower in patients with GEJA than in patients with distal gastric cancer (80.7 vs. 94.6%, p < 0.05). Concurrent H. pylori infection and corporal gastritis were not observed in patients with Barrett's esophageal cancer. CONCLUSION: Our study demonstrated that GEJA has 2 etiologic types; one of these types is associated with H. pylori infection and resembles distal gastric cancer, and the other one is not associated with H. pylori infection or Barrett's esophageal cancer.

Expression of sphingosine-1-phosphate receptor 1 in mantle cell lymphoma.

The distribution and pathological significance of sphingosine-1-phosphate receptor 1 expression are still unclear. In this study, we evaluated sphingosine-1-phosphate receptor 1's suitability as a diagnostic marker for malignant lymphoma by immunostaining formalin-fixed paraffin-embedded sections using a well-defined commercial anti-sphingosine-1-phosphate receptor 1 antibody. Sphingosine-1-phosphate receptor 1 was strongly expressed on the surface of small lymphocytes forming primary lymphoid follicles and in the mantle zone of secondary lymphoid follicles. Microarray-based immunohistochemistry with tissue samples from 85 lymphoid malignancy cases demonstrated that sphingosine-1-phosphate receptor 1 was expressed on the surface of mantle cell lymphoma cells. Strong expression was observed in all classical mantle cell lymphoma cases involving the lymph node (19 out of 19), gastrointestinal tract (10 out of 10), bone marrow (9 out of 9), and orbita (1 out of 1). Good results were obtained even in sections where cyclin D1 signals were lost because of over-fixation and/or decalcification. One aggressive variant of mantle cell lymphoma displayed a weaker membranous staining than classical mantle cell lymphoma in the lymph node and bone marrow. In a cyclin D1-negative mantle cell lymphoma of the orbita, no conclusive result was obtained. No cases of follicular lymphoma, marginal zone lymphoma, B lymphoblastic leukemia/lymphoma, or Burkitt's lymphoma showed any significant expression, whereas 2 out of 6 chronic lymphocytic leukemia/small lymphocytic lymphomas in bone marrow, 1 out of 3 lymphoplasmacytic lymphomas in the lymph node, and 2 out of 37 diffuse large B-cell lymphomas exhibited staining. A quantitative reverse transcription polymerase chain reaction-based analysis of mantle cell lymphoma lines revealed the sphingosine-1-phosphate receptor 1 mRNA expression level to be well correlated with the results of immunocytochemistry, flow cytometry, and western blotting. Thus, sphingosine-1-phosphate receptor 1 immunohistochemistry may be useful in the histological diagnosis of mantle cell lymphoma with formalin-fixed and paraffin-embedded sections. The antigen may be particularly valuable in cases where cyclin D1 immunostaining is not successful.

Pancreatic serous microcystic adenoma with extensive oncocytic change.

Herein is reported a case of pancreatic serous microcystic adenoma with extensive oncocytic change in a 73-year-old woman. Histologically the tumor consisted of numerous small cysts, separated by thin or broad fibrous septa. These cysts were lined with uniform cells having abundant eosinophilic granular cytoplasm, which was negatively or weakly stained with PAS. Immunohistochemically, the cyst-lining cells were positive for cytokeratin (CK) 7, CK19, MUC1, MUC6, alpha-inhibin, and neuron-specific enolase (NSE), and negative for CK8, CK20, MUC2, and MUC5AC; these immunoprofiles coincide with those of serous microcystic adenoma. Immunostaining with anti-mitochondrial antibody showed dense granular positivity in the cytoplasm, which suggested an oncocytic phenotype. Thus, this case is considered a variant of serous microcystic adenoma characterized by extensive oncocytic change. To the authors' knowledge no similar case has been reported in the literature. It may pose problems in the differential diagnosis of the cystic pancreatic tumors with oncocytic change, but can be diagnosed on histology and immunohistochemistry.

Transformation from follicular lymphoma to high-grade B-cell lymphoma/leukemia with additional t(2;8)(p12;q24), with inverse expressions of c-MYC and BCL-2, and light-chain switch.

Transformation from follicular lymphoma (FL) to high-grade B-cell lymphoma/leukemia (BLL) has been reported in patients with additional translocations involving the c-MYC gene. The previously reported cases were related to t(8;14) and t(8;22) but not to t(2;8). Herein is reported an FL that terminated in BLL following additional t(2;8). In accordance with previous reports, increased expression of c-MYC was observed in the present case but, interestingly, BCL-2 expression was inversely decreased after the transformation. In addition, the cell-surface immunoglobulin light-chain of lymphoma cells was initially kappa type and was then gradually replaced with the lambda type after transformation. Downregulation of BCL-2 and light-chain switch have rarely been reported in previous cases of FL transformation involving c-MYC, suggesting that additional t(2;8) translocation may play a role in these events.

Rosai-Dorfman disease presenting as a solitary mediastinal mass.

Rosai-Dorfman disease (RDD) involving an extranodal site is a diagnostic challenge. Reported herein is the case of a 67-year-old man who presented with a solitary superior mediastinal mass. The lesion was clinically suspected of malignancy including lymphoma because of its high uptake during a (67)Ga-scintigram and (18)F-fluorodeoxyglucose-positron emission tomography. There was no evidence of spread of the disease. Histology of thoracoscopic biopsy specimens indicated granulomatous lesion with infiltration of lymphocytes, plasma cells, and histiocytes with lymphocytes engulfed in their cytoplasm. The lesion did not contain lymph node or thymic elements. On immunohistochemistry the histiocytes were positive for S-100 protein, CD68, and CD163 but were negative for CD1a. These findings suggested a diagnosis of RDD. Despite lack of intervention, the lesion remained almost the same size for 3 years. To the best of the authors' knowledge this is the first case of RDD presenting as a solitary mediastinal mass.

Successful Cord Blood Transplantation in a Werner Syndrome Patient with High-risk Myelodysplastic Syndrome.

Werner syndrome (WS) confers a high risk of the development of neoplasias, including hematological malignancies, and curative treatment for these malignancies is difficult to achieve. A 44-year-old man with myelodysplastic syndrome was admitted to our hospital. He was diagnosed with mutation-proven WS. He underwent cord blood transplantation (CBT) following fludarabine, busulfan, and melphalan administration. A chimerism analysis of his marrow blood on day 62 showed a donor pattern >95%, which confirmed engraftment. The patient lived for 15 months while maintaining remission of MDS without treatment-related toxicity. Our case shows that CBT can be a treatment modality for WS patients with hematological malignancies.

Novel neuroblastoma amplified sequence (NBAS) mutations in a Japanese boy with fever-triggered recurrent acute liver failure.

Biallelic mutations in the neuroblastoma amplified sequence (NBAS) gene have been reported to cause two different clinical spectra: short stature with optic nerve atrophy and Pelger-Huët anomaly (SOPH) syndrome and infantile liver failure syndrome 2 (ILFS2). Here, we describe a case of a 3-year-old Japanese boy who presented with fever-triggered recurrent acute liver failure (ALF). The clinical characteristics were considerable elevation of liver enzymes, severe coagulopathy, and acute renal failure. In addition to the liver phenotype, he had short stature and Pelger-Huët anomaly in the peripheral granulocytes. Whole-exome and Sanger sequencing of the patient and his parents revealed that he carried novel compound heterozygous missense mutations in NBAS, c.1018G>C (p.Gly340Arg) and c.2674 G>T (p.Val892Phe). Both mutations affect evolutionarily conserved amino acid residues and are predicted to be highly damaging. Immunoblot analysis of the patient's skin fibroblasts showed a normal NBAS protein level but a reduced protein level of its interaction partner, p31, involved in Golgi-to-endoplasmic reticulum retrograde vesicular trafficking. We recommend NBAS gene analysis in children with unexplained fever-triggered recurrent ALF or liver dysfunction. Early antipyretic therapy may prevent further episodes of ALF.

Immunohistochemical detection of sphingosine-1-phosphate receptor 1 in vascular and lymphatic endothelial cells.

Sphingosine-1-phosphate receptor 1 (S1P(1)), a receptor for sphingosine-1-phosphate, has been shown to play an important role in the migration, proliferation, and survival of several types of cell including endothelial cells. Given that S1P(1) signaling could serve as a therapeutic target, we evaluate the expression of S1P(1) in formalin-fixed and paraffin-embedded sections from human tissues, using automated immunostainers (Ventana). The specificity of the polyclonal rabbit anti-human S1P(1) antibody used in this study was defined by immunostaining of the vasculature in S1P ( 1 ) ( -/- ) and S1P ( 1 ) ( +/- ) mouse embryos. The antibody stained the newly formed vasculatures ex vivo in a serum-free matrix culture model using rat aortic rings. In human specimens, S1P(1) was strongly expressed on the cell surface membrane of endothelial cells of blood and lymphatic vessels in all tissues examined. The expression of S1P(1) was confirmed by the flow cytometric analysis and real time RT-PCR of an angiosarcoma cell line. This study indicates that S1P(1) can be used as an immunohistochemical marker for human tissue endothelial cells.

Small cell variant of anaplastic large cell lymphoma diagnosed by a novel chromosomal abnormality t(2;5;3)(p23;q35;p21) of bone marrow cells.

Because of the rarity and the morphological variations, small cell variant of anaplastic large cell lymphoma (ALCL) represents a diagnostic challenge. Herein is reported a case of leukemic type of small cell variant of ALCL, in which the diagnosis was established by a cytogenetic analysis. The patient was a 23-year-old woman who presented with fever and leukocytosis with circulatory atypical lymphoid cells. The initial differential diagnosis on bone marrow trephine biopsy sections included viral infection and peripheral T-cell lymphoma unspecified. But a cytogenetic study on bone marrow cells indicated a novel complex translocation, t(2;5;3)(p23;q35;p21), which led to confirmation of anaplastic lymphoma kinase (ALK)-positive pleomorphic small to medium-sized cells scattered in bone marrow cells, on immunohistochemistry. ALK was distributed in both nuclear and cytoplasmic regions of neoplastic cells. The patient achieved complete remission after four courses of combination chemotherapy, and received autologous peripheral stem cell transplantation (auto-PBSCT) after two additional courses of combination chemotherapy, but relapsed 2 months after auto-PBSCT in the bilateral lung. Allogeneic stem cell transplantation led to a second remission. This case demonstrates the diagnostic importance of cytogenetic study for malignant lymphoma involving bone marrow.

CD8+, CD56+ (natural killer-like) T-cell lymphoma involving the small intestine with no evidence of enteropathy: clinicopathology and molecular study of five Japanese patients.

The present study reports five CD8+, CD56+ (natural killer (NK)-like) T-cell lymphomas involving the small intestine without evidence of enteropathy, from Japan. Three were intestinal T-cell lymphoma. The site of origin of the other two was not definitive. Four of five patients underwent emergency operation because of intestinal perforation. The small intestines of these patients had multiple ulcerative lesions with or without demarcated tumors. Histologically, the lymphoma cells were monomorphic or slightly pleomorphic and displayed epitheliotropism of varying degrees. Lymphoma cells of all patients shared the common phenotype: CD3+, CD4-, CD5-, CD8+, CD56+, CD57-, T-cell intracellular antigen-1+, granzyme B+. In contrast to nasal/nasal type NK-cell lymphomas, they had clonal rearrangement of T-cell receptor(TCR) genes and were negative for EBV-encoded RNA. Immunohistochemistry and genetics suggested that three cases were of alpha beta T-cell origin and two cases were of gamma delta T-cell origin. There was no evidence of enteropathy in any patient. The cases followed a clinically aggressive course with a frequent involvement of lung. According to the classification based on the recent genetic studies of European enteropathy-type intestinal T-cell lymphoma (ETL), the present cases could be classified as type 2 ETL.

[Clinical analysis of eight patients with primary follicular lymphoma in the duodenum].

We performed a clinical analysis on 8 patients with primary follicular lymphoma in the duodenum taken from among 26 cases of primary gastrointestinal malignant lymphoma treated in our division. The median age was 60 years (range 48 to 82 yr). The ratio of males to females was 4:4. The chief complaints were no symptoms in 4 cases, heartburn in 2 cases, lower abdominal pain in 1 case, and back pain in 1 case. All patients were in clinical stage I EA. Gastroendoscopic findings showed multiple whitish granules around the ampulla of Vater in all patients. Involvement of the site in 6 cases was only located at the second portion; lesions in the other 2 cases were located at the second portion, and at the third portion or fourth portion, respectively. A histological study showed follicular lymphoma grade 1, and an immunohistological study demonstrated that the lymphoma cells were positive for CD79a, CD10, CD20, and bcl-2. Five patients were positive for the FISH analysis fusion signal of IgH/bcl-2 genes. Rituximab with CHOP therapy was performed for 7 patients. Seven patients are currently alive, and one died of uterine cancer. At the medium-term 39 month-follow-up, 7 patients were in complete remission, and 1 patient was in partial remission. Rituximab with CHOP (CVP) therapy is a possible treatment for primary follicular lymphoma in the duodenum. Further consideration of appropriate therapy for this disease might be necessary.

Measurement of the thickness of the fibrous cap by optical coherence tomography.

BACKGROUND: Identification of the fibrous cap is important because its thickness is a major determinant of plaque vulnerability in lipid-rich plaque. Thus, a high-resolution imaging technique may be a promising method for the identification of the fibrous cap within lipid-rich plaque. The purpose of this study was to investigate the feasibility of using optical coherence tomography (OCT) to measure the thickness of the fibrous cap within lipid-rich plaque. METHODS AND RESULTS: We examined 35 lipid-rich plaques from 102 coronary arterial segments of 38 human cadavers (22 men and 16 women; mean ages, 74 +/- 7 years). Optical coherence tomography and corresponding histological images were digitized for measurement of the thickness of fibrous cap, and the results between OCT and histological examination were compared. There was good correlation of the thickness of the fibrous cap between OCT and histological examination (y = 0.97x + 28.49; r = 0.90; P < .001). A Bland-Altman test showed good agreement of the thickness of the fibrous cap between OCT and histological examination (mean difference, -24 +/- 44 microm). CONCLUSIONS: Optical coherence tomography provides an accurate representation of the thickness of the fibrous cap and may prove useful in assessing plaque vulnerability in lipid-rich plaque.

Assessment of coronary arterial plaque by optical coherence tomography.

The purpose of this study was to analyze the ability of optical coherence tomography (OCT) to identify coronary arterial plaque diagnosed by histologic examination. We examined 166 sections from 108 coronary arterial segments of 40 consecutive human cadavers (24 men and 16 women; mean age 74 +/- 7 years). The plaque type was classified as fibrous (n = 43), fibrocalcific (n = 82), or lipid-rich (n = 41). The accuracy of OCT and intravascular ultrasound (IVUS) in characterizing the plaque type was studied, with the histologic consensus diagnosis serving as the gold standard. OCT, as well as IVUS, had high sensitivity and specificity for characterizing the different types of atherosclerotic plaque. OCT had a higher sensitivity for characterizing lipid-rich plaques than IVUS (85% vs 59%, p = 0.03). In conclusion, the high resolution of OCT permitted evaluation of lipid-rich plaques more accurately than IVUS.

Assessment of coronary arterial thrombus by optical coherence tomography.

We analyzed optical coherence tomographic (OCT) characteristics of different types of coronary thrombi that had been confirmed at postmortem histologic examination. We examined 108 coronary arterial segments of 40 consecutive human cadavers. OCT images of red and white thrombi were obtained and the intensity property of these thrombi was analyzed. Red and white thrombi were found in 16 (17%) and 19 (18%) of the 108 arterial segments, respectively. Red thrombi were identified as high-backscattering protrusions inside the lumen of the artery, with signal-free shadowing in the OCT image. White thrombi were identified as low-backscattering projections in the OCT image. There were no significant differences in peak intensity of OCT signal between red and white thrombi (130+/-18 vs 145+/-34, p=0.12). However, the 1/2 attenuation width of the signal intensity curve, which was defined as the distance from peak intensity to its 1/2 intensity, was significantly different between red and white thrombi (324+/-50 vs 183+/- 42 microm, p<0.0001). A cut-off value of 250 microm in the 1/2 width of signal intensity attenuation can differentiate white from red thrombi with a sensitivity of 90% and specificity of 88%. We present the first detailed description of the characteristics of different types of coronary thrombi in OCT images. Optical coherence tomography may allow us not only to estimate plaque morphology but also to distinguish red from white thrombi.

Hypoxia-inducible factor-1 drives the motility of the erythroid progenitor cell line, UT-7/Epo, via autocrine motility factor.

OBJECTIVE: It is well known that hypoxic stress strongly enhances erythropoiesis, but the effect of hypoxia on erythroid progenitors has not been examined precisely. In the present study, using the erythropoietin-dependent cell line UT-7/Epo, which has characteristics of erythroid progenitors, we investigated a novel role of hypoxia in erythropoiesis. METHODS: UT-7/Epo and four other hematopoietic and lymphoid cell lines (HL-60, THP-1, Raji, and CEM) were cultured in 20%, 5%, or 1% O2. Morphology was observed under a phase-contrast microscope. Cell motility was evaluated using the Transwell migration assay. An analysis of the protein level of hypoxia-inducible factor-1 (HIF-1) alpha and autocrine motility factor (AMF) was conducted using Western blotting and immunocytochemistry, respectively. Reverse transcription polymerase chain reaction was performed to evaluate the expression of AMF mRNA. Human bone marrow stromal cells were used in cocultures with UT-7/Epo. Apoptosis of UT-7/Epo was examined by immunocytochemistry using an antiactive form of caspase 3 antibody. RESULTS: Among the five cell lines, UT-7/Epo exhibited active pseudopodial extension in hypoxia (1% O2), and cell motility was increased. HL-60, THP-1, Raji, and CEM did not show an increase in cell motility even in 1% O2. In addition, expression of the alpha-subunit of HIF-1 was activated by hypoxia, and expression of the mRNA and protein of AMF induced by HIF-1, increasing cell motility, was promoted. The addition of an HIF-1 inhibitor, cadmium chloride (CdCl2), or alpha-ketoglutarate (2-oxoglutarate) decreased the AMF mRNA expression, and an AMF inhibitor, erythrose 4-phosphate, decreased the cell motility. When UT-7/Epo was cocultured with human bone marrow-derived stromal cells that significantly inhibit the apoptosis of UT-7/Epo, the migration of UT-7/Epo under the stromal cells (pseudoemperipolesis) was increased in hypoxia. CONCLUSION: Under hypoxic conditions, erythroid progenitors may exhibit active migration in the bone marrow and the opportunity for contact with stromal cells increases, inhibiting apoptosis.

Contrast-enhanced ultrasonography with Sonazoid for diagnosis of gangrenous cholecystitis.

PURPOSE: This prospective study investigated the ability of contrast-enhanced ultrasonography (CEUS) with Sonazoid to diagnose gangrenous cholecystitis and determined the inter-observer agreement. METHODS: From September 2012 to August 2014, 27 patients with acute cholecystitis underwent preoperative CEUS (registration number 1277). After Sonazoid injection, harmonic imaging of the gallbladder wall was performed, and the findings were recorded using movie clips. The signal intensity was classified as absence (uncomplicated) or presence of perfusion defects (gangrenous). The physician performing CEUS recorded the findings immediately after the examination. Another physician (blinded to the clinical information) then reviewed the movie clips and recorded the findings. The final diagnosis was determined by histological examination in all 27 patients. RESULTS: The final diagnosis was gangrenous cholecystitis in 15 patients and uncomplicated cholecystitis in 12. On CEUS examination, perfusion defects were detected in 10 patients with gangrenous cholecystitis, giving a sensitivity of 66.7 %, specificity of 100 %, positive predictive value of 100 %, and negative predictive value of 70.6 %. On review of the movie clips, these values were 73.3, 100, 100, and 75.0 %, respectively. The inter-observer agreement between physicians was good (κ coefficient, 0.64). CONCLUSIONS: CEUS with Sonazoid is a useful and reproducible modality for diagnosing gangrenous cholecystitis.