Megakaryopoiesis and platelet function in polycythemia vera and essential thrombocythemia patients with JAK2 V617F mutation.

Patients with Ph chromosome negative myeloproliferative disease (Ph-MPD) have an increased risk of vascular complications. It remains controversial whether patients with the JAK2 V617F mutation (V617F) exhibit increased risk, while recent growing evidence has shown a critical role for V617F in clonal erythropoiesis in Ph-MPD. We studied 53 patients with Ph-MPD especially in relation to megakaryopoiesis, the thrombotic complications and the presence of V617F. Using novel mutation-specific PCR which is a highly sensitive PCR-based assay for detection of JAK2 mutated allele(s), we identified V617F in 38 Ph-MPD, which include 13 polycythemia vera (PV), 23 essential thrombocythemia (ET) and 2 chronic idiopatic myelofibrosis. The numbers of megakaryocytes were significantly increased in PV and ET patients with V617F, but the platelet counts were slightly lower. Although statistically not significant, the incidence of thrombotic events was higher in the group with V617F compared to in those without the mutation. Agonist-induced in vitro platelet aggregation and platelet adhesion were not affected by the presence of this mutation. Nonetheless, we found a hypercoagulable state in Ph-CMPD with V617F by employing whole blood thromboelastography. It suggests pre-thrombotic tendencies in CMPD are complex and JAK2 V617F mutation might have a role in vivo blood coagulation by altering not only the number, but function(s) of all three myeloid cells, including red blood cells, white blood cells and platelets in Ph-CMPD.

Histopathology of bone marrow reconstitution after umbilical cord blood transplantation for hematological diseases.

To study hematopoietic reconstitution in umbilical cord blood transplantation (CBT), bone marrow (BM) histology was investigated in 35 biopsies after bone marrow transplantation (BMT) and in 40 biopsies after CBT. BM biopsies were obtained at different times after transplantation and were evaluated for cellularity, number of megakaryocytes and CD34-positive cells, and fibrosis. In biopsies up to 29 days after BMT, cellularity was increased and megakaryocytes were observed, but at 29 days after CBT, biopsies showed severe cellular depletion and almost no megakaryocytes. In addition, fewer CD34-positive cells were observed after CBT compared to after BMT. After day 30 after CBT, hematopoietic recovery of the BM was gradually observed and after day 100 after transplantation, no essential differences were observed between BMT and CBT. Hematopoietic recovery of the BM after CBT was delayed compared to that after BMT, but engraftment of donor cells after CBT was also observed in histopathologically. To the best of the authors' knowledge this is the first histopathological description of BM reconstitution after CBT.

[Splenic irradiation as a successful treatment for an elderly patient with B-cell prolymphocytic leukemia].

We report a case of B-cell prolymphocytic leukemia (B-PLL) that was treated successfully with splenic irradiation (SI). An 86-year-old man underwent a medical examination for lumbago and general fatigue at another hospital in June 2007. A compressed lumbar fracture and splenomegaly were found using computed tomography (CT). Thereafter, the patient consulted our hospital because of leukocytosis. Peripheral blood showed hemoglobin level 9.8 g/dl and white blood cell count 38.1x10(9)/l with 91% atypical cells. Surface marker analysis demonstrated that atypical cells were positive for CD20, CD22, FMC7, surface IgM, surface IgD and kappa, but were negative for CD5, TdT and lambda. The morphology of these cells was compatible with prolymphocytes with prominent nucleoli and condensed nuclear chromatin. A diagnosis of B-PLL was made. SI (total dose 20 Gy) was chosen for the treatment and a single course of SI was very effective without causing any significant adverse events. This case demonstrates that SI may remain valuable for the treatment of B-PLL in an elderly patient.

[Close correlations between CD20 expression, a small mature plasma cell morphology and t(11 ; 14) in multiple myeloma].

Stratification of patients with multiple myeloma (MM) may be important. We investigated 138 MM patients, focusing on correlations between CD20 expression, 11 ; 14 translocation, morphology of MM cells, cyclin D1 immunostaining, and the prognosis. About 15% of patients (7/47cases) were CD20-positive, small mature MM cells, with positive cyclin D1 in the nucleus and 11; 14 translocation. Two color analysis of CD38 x CD20 antigens may be necessary to investigate CD20 expression on MM cells. Rituximab may be effective for the treatment of CD20-positive MM.

Prevention of perinatal hepatitis B virus transmission by combined passive-active immunoprophylaxis in Iwate, Japan (1981-1992) and epidemiological evidence for its efficacy.

In a model area in Iwate, Japan, with a population of 1.4 million, the immunoprophylaxis of perinatal transmission of hepatitis B virus (HBV) was started in 1981 and covered >60% of all births already in 1986 when it became mandatory by the national program. Babies born to mothers who carried hepatitis B surface antigen (HBsAg) along with hepatitis B e antigen (HBeAg) in serum received hepatitis B immune globulin (HBIG) at birth and 2 months as well as vaccine at 2, 3 and 5 months after birth. In 1985, 39 of 45 (86.7%) babies who received immunoprophylaxis did not develop the HBV carrier state. During 1986-1992, 100286 of 104493 (96.0%) expecting mothers received tests for HBsAg, and it was detected in 1242 (1.2%) of them. Among the mothers carrying HBsAg, 257 (20.7%) were positive for HBeAg and their babies received immunoprophylaxis. Reflecting effects of immunoprophylaxis, the prevalence of HBsAg decreased from 0.75% (78/10437) in the children born during 1978-1980 to 0.23% (46/20812) in those during 1981-1985 (P<0.001), and further to 0.04% (12/32049) in those during 1986-1990 (P<0.001). The prevalence rates of antibody to HBsAg (anti-HBs) were 1.52, 0.79 and 0.85% in the three groups of children (P<0.001 between those during 1978-1980 and the others). The frequency of antibody to HBV core in the children with anti-HBs diminished remarkably from 76.7% (23/30) in those born in 1971 to 9.0% (6/67) in those born in 1990, thereby indicating a marked decrease in resolved infection and increase in acquired immunity to HBV as the results of immunoprophylaxis.

[Acute myeloid leukemia originating from the same leukemia clone after the complete remission of acute lymphoid leukemia].

A 22-year-old female was diagnosed as having acute lymphoid leukemia (ALL) in February 1995, from the findings of peroxidase negative, CD10+, CD19+, TdT+ and rearrangement of IgH and TCR beta. AdVP (doxorubicin, vincristine and prednisolone) therapy achieved a complete remission (CR). Bone marrow transplantation had to be abandoned because of the lack of an HLA-identical donor. Intensification therapy was thus carried out repeatedly. In June 1998, myeloblast with Auer rods, peroxidase positive, CD13+, CD33+ and HLA-DR+, appeared. The patient was diagnosed as having lineage switch acute myeloid leukemia (AML) from ALL. Though A-DMP (cytosine arabinoside, daunorubicin, 6-mercaptopurine) therapy was resistant, AdVP therapy led to a CR. The patient died of cardiotoxicity from anthracyclines in February 1999. From the results of the Ramasamy method using the clonal rearrangements of the Ig heavy chain gene locus, the origin of the pathological cells of ALL and AML was indicated to be the same leukemia clone.

[Severe type of Epstein-Barr virus associated hemophagocytic syndrome successfully treated with T-COP-E and splenectomy].

A 16-year-old girl was admitted to our hospital because of high fever, abdominal pain, and jaundice. Abnormal lymphocytes and hemophagocytic cells had infiltrated the bone marrow. Laboratory data revealed a severe type of hemophagocytic syndrome accompanied by an initial Epstein-Barr virus (EBV) infection. Persistent EBV infection was identified by polymerase chain reaction (PCR) detection of EBV-DNA in peripheral blood and bone marrow mononuclear cells. The limited efficacy of initial treatment with high-dose gamma-globulin, plasmapheresis, and high-dose methylprednisolone prompted us to administration of T-COP-E (VP-16). Two courses of T-COP-E improved the patient's clinical symptoms and laboratory data; however, marked splenomegaly remained. In addition, fever and serum increase of lactate dehydrogenase (LDH) and cytokines such as gamma-interferon recurred shortly after chemotherapy. On day 53 after diagnosis, the patient underwent laparoscopic splenectomy. The resected spleen weighted 420 g and abnormal lymphocytes in the spleen were positive for CD 8 and negative for CD 56. In situ hybridization revealed EBV-encoded small RNAs (EBERs) in the abnormal lymphocytes. Clinical symptoms including high fever disappeared shortly after the splenectomy, and laboratory data returned to normal. Lymphocytosis after the splenectomy was not observed. We continued out patient monitoring of the case, and 16 months after diagnosis, EBV-DNA in peripheral blood mononuclear cells was not detected, even by PCR.

Identification of Human Intestinal Microbiota of 92 Men by Data Mining for 5 Characteristics, i.e., Age, BMI, Smoking Habit, Cessation Period of Previous Smokers and Drinking Habit.

The intestinal microbiota compositions of 92 men living in Japan were identified following consumption of identical meals for 3 days. Fecal samples were analyzed by terminal restriction fragment length polymorphism with 4 primer-restriction enzyme systems, and the 120 obtained operational taxonomic units (OTUs) were analyzed by Data mining software focusing on the following 5 characteristics, namely, age, body mass index, present smoking habit, cessation period of previous smokers and drinking habit, according to the answers of the subjects. After performing Data mining analyses with each characteristic, the details of the constructed Decision trees precisely identified the subjects or discriminated them into various suitable groups. Through the pathways to reach the groups, practical roles of the related OTUs and their quantities were clearly recognized. Compared with the other identification methods for OTUs such as bicluster analyses, correlation coefficients and principal component analyses, the clear difference of this Data mining technique was that it set aside most OTUs and emphasized only some closely related ones. For example for a selected characteristic, such as smoking habit, only 7 OTUs out of 120 were able to identify all smokers, and the remaining 113 OTUs were thought of as data noise for smoking. Data mining analyses were affirmed as an effective method of subject discrimination for various physiological constitutions. The species of bacteria that were closely related to heavy smokers, i.e., HaeIII-291, were also discussed.

Primary pulmonary classical hodgkin lymphoma with two recurrences in the mediastinum : a case report.

We report a case of primary pulmonary classical Hodgkin lymphoma (CHL) in a 58-year-old woman. Twelve years ago, the patient complained of slight fever and weight loss. A mass of about 5 cm in diameter was seen in the right lung on radiography and computed tomography (CT). Right total pneumonectomy and resection of mediastinal lymph nodes were performed. A pathological examination led to a strong suspicion of Hodgkin disease (HD) (now referred to as CHL), but a definite diagnosis could not be made at the time. Six years later, a chest CT showed a tumor around the ascending aorta, which was treated successfully by radiation therapy. Six years later, the chest CT revealed a tumor in the anterior mediastinum. CHL was diagnosed based on an immunohistochemical re-examination of lung specimens resected 12 years earlier and CT-guided fine needle tumor biopsy specimens of the second recurrent tumor in the anterior mediastinum were compatible with the recurrence of CHL. Therefore, we diagnosed this case as primary pulmonary CHL that later relapsed in the mediastinum. The tumor size was reduced by radiation therapy and the patient is currently under observation as an outpatient.

Clinical implications of abnormalities of chromosomes 1 and 13 in multiple myeloma.

Stratification of patients with multiple myeloma according to clinical severity was attempted by chromosomal analysis of 180 bone marrow specimens from 79 patients. The 79 patients were hospitalized and treated between 1994 and 1999. Abnormalities of chromosome 1 were detected at the initial medical examination in 8 (10%) of the 79 patients and were found during follow-up in additional 3 patients. Abnormalities of chromosome 1 were often detected in IgA (4/17) and IgD (2/4) multiple myeloma, while detection in IgG myeloma was relatively rare (4/42). The relevant break points were 1q12 in 5 patients and 1q42 and 1q21 in 3 patients, while 3 patients had trisomy 1. Deficiency of the long arm of chromosome 13 was detected in 6 patients, and this was believed to imply a prognosis. The long arm was completely deleted in 2 patients and part of the arm (13q10-14) was deleted in 4 patients. It is interesting that all 6 patients had concomitant abnormalities of chromosome 1. Although the initial response to treatment of patients having abnormal chromosomes 1 and 13 was comparable to that of patients having other chromosomal abnormalities or patients who were karyotypically normal, the median survival time was only 18 months (p < 0.02). Consequently, aggressive treatment such as early stem cell transplantation and so on is needed to improve their survival.