Establishment of a GIS monitoring system for schistosomiasis japonica in Kofu, Japan.

Although the incidence of human infection with Schistosoma japonicum in Japan fell to zero in 1977, the threat of the possible re-emergence of the disease caused by this trematode still exists. Surveillance of the parasite's intermediate host, Oncomelania nosophora, in Kofu basin therefore began in 1996. A simple, new method for monitoring O. nosophora in an at-risk area in Kofu, which is based on a geographical information system (GIS), was established. At each monitoring site (of which there were 120 from 1996 until 2000, and 60 from 2001 until 2003), the O. nosophora in two quadrats, each measuring 25 x 25 cm, were collected. During the study, the exact location of each site was determined using a hand-held global-positioning system (GPS). This allowed all the sites to be digitally mapped, so that anyone with a hand-held GPS could and can reach each site. The snail and location data were processed using commercial GPS/GIS software packages and used to create a risk map for schistosomiasis re-emergence. Although all snails collected between 1996 and 2003 were uninfected, the proportion of investigated sites in which O. nosophora was detected increased from 36.7% in 1996 to 56.7% in 2003. The mean number of O. nosophora collected per snail-positive site fluctuated widely, between 8.2 and 57.4, in each calendar year. Over the study period there appeared to be a shift southwards in the areas with high densities of O. nosophora. The present results indicate that it is possible to utilize a GIS-based method for the long-term monitoring of the possible re-emergence of schistosomiasis japonica in Japan.

PAX 8 regulates human WT1 transcription through a novel DNA binding site.

The Wilms' tumor gene (WT1) is an essential gene for kidney and gonadal development, although how WT1 expression is induced in these tissues is not known. One kidney transcription factor likely to play a role in this regulation is PAX 8. The co-expression of WT1 and PAX 8 during kidney development and in Wilms' tumors with an epithelium predominant histology suggested a possible interaction, and indeed, we identified potential core PAX-binding sites in the WT1 promoter. Endogenous PAX 8 plays an important role in the activation of the WT1 promoter, since promoter activity is much stronger in cells with PAX 8 than without. Using binding assays, we searched for evidence of PAX 8-DNA interactions throughout the 652-base pair human WT1 promoter and found only one functional PAX 8 site with DNA binding activity, located 250 base pairs 5' of the minimal promoter. The responsiveness of the PAX 8 site was confirmed by assessing its ability to function as an enhancer significantly activating the minimal promoter in a position- and orientation-independent manner. Using transfection assays, we demonstrated that either endogenous or exogenously added PAX 8 activated the WT1 promoter and that this promoter up-regulation depended upon the presence of an intact PAX 8-binding site. In contrast, the previously reported core PAX 8-binding sites identified by computer analysis of the WT1 promoter failed to specifically bind in vitro translated PAX 8 protein or activate the minimal promoter. Thus, we identified a novel functional binding site for the transcription factor PAX 8, suggesting that part of its role in kidney development may be as a modulator of WT1 expression in the kidney.

The Wilms' tumor gene WT1 can regulate genes involved in sex determination and differentiation: SRY, Müllerian-inhibiting substance, and the androgen receptor.

Genital abnormalities associated with Wilms' tumors in the WAGR and Denys-Drash syndromes and the failure of the gonads to develop in Wilms' tumor gene (wt1)-homozygous mutant mice suggest that WT1 may also function in sexual development. To elucidate the mechanism of action of WT1 in embryonal sexual development, we examined how the four isoforms of WT1 regulate the transcription of several genes involved in sexual development using cotransfection assays. SRY (the sex-determining region of the Y chromosome) promoter was strongly activated by the WT1 isoforms without the KTS tripeptide, WT1(-)KTS, but was not activated by the WT1 isoforms with the KTS tripeptide, WT1(+)KTS, in all cells tested. The second alternative splicing site, which inserts the tripeptide KTS, alters the DNA binding capability. The MüAdullerian-inhibiting substance (MIS) promoter was strongly repressed by WT1(-)KTS isoforms and more weakly repressed by the WT1(+)KTS isoforms in Sertoli cells but not in HeLa cells. The androgen receptor (AR) promoter was strongly repressed by the WT1(-)KTS isoforms in all cells tested and was more weakly or not repressed by WT1(+)KTS isoforms depending on cell lines. Electrophoretic mobility shift assays showed strong binding by recombinant WT1(-)KTS protein and weaker or no binding by the WT1(+)KTS protein to DNA probes containing WT1 binding sites from these three promoters. The results of these functional and binding assays suggest that WT1 has an important role in regulation of genes involved in embryonal sexual development and that WT1 can function as a transcriptional activator.

[A patient with liver cirrhosis manifesting various symptoms including cerebellar ataxia due to germanium intoxication].

A patient with hepatitis B virus-associated cirrhosis manifested various symptoms such as anemia, renal damage and neurological signs including cerebellar ataxia due to long-term administration of germanium-containing food. The patient was a 40-year-old male who had taken germanium containing mineral cheese for 26 months after he was diagnosed as having cirrhosis. Twenty four months after beginning to take the mineral cheese, he began manifesting paresthesia of the extremities, dysarthria and gait ataxia. Laboratory findings revealed anemia and renal damage. Biopsy of the peripheral nerve revealed loss of the large sheathed nerve, a characteristic feature of germanium intoxication. A high concentration of germanium (GeO2) was detected in patient's hair and urine. Cerebellar ataxia was characteristic in this patient, which was not reported in the previous papers.

[An autopsy case of citrullinemia type II complicated with chronic pancreatitis].

A 34-year-old woman had developed frequent episodes of disorientation at night since a year ago. Her blood ammonia level was found to be markedly increased. Serum amino acid pattern and biochemical analysis of urea cycle enzymes in the liver indicated type II citrullinemia. Pancreatolithiasis was found by ultrasonography and CT scan. After a short remission following treatment by protein restriction and oral administration of sodium benzoate, she had disturbed consciousness due to exaggerated hyperammonemia and died of severe brain edema. In this report, efficacy of the treatment for hyperammonemia and histopathological findings are discussed.

Assessment of abdominal involvement of adult T-cell leukemia/lymphoma by ultrasonography: comparison among four clinical types.

Adult T-cell leukemia/lymphoma (ATLL) is an HTLV-I associated lymphoid malignancy frequently seen in Japan. Abdominal involvement in 40 patients with ATLL were assessed by ultrasonography and the findings seen in four clinical types, acute, chronic, lymphoma and smoldering, were compared. Splenomegaly was frequently found in the cases of acute and lymphoma types, and the sizes of the spleens measured by ultrasonography correlated well with the disease activity. Hepatomegaly was also found more frequently in acute and lymphoma types, and hepatosplenomegaly was proved to be due to the infiltration by ATL cells. Nodular lesions in spleen and liver and abdominal lymph node swelling were also found frequently in the lymphoma type but rarely in the other types. Ascites, pleural effusion, and pericardial effusion were found in the active stage of acute and lymphoma types. Ultransonography also could detect findings associated with therapies. Thus, ultrasonography studies were found to be very useful for assessing the clinical classification, examining various pathological conditions associated with ATLL, and monitoring the disease activity.

Ultrasonographic studies on abdominal complications in patients receiving marrow-ablative chemotherapy and bone marrow or blood stem cell transplantation.

Abdominal complications were evaluated with ultrasonography in 20 patients who received marrow-ablative chemotherapy and bone marrow or blood stem cell transplantation for the treatment of hematologic malignancies. Ultrasonographic findings compatible with veno-occlusive disease of the liver, cytomegalovirus infection of the colon, hepatic lesion of graft-versus-host disease, and cyclophosphamide-induced hemorrhagic cystitis were demonstrated in 6 of these patients. In addition, ascites, pleural effusion, gall bladder wall thickening, and hepatosplenomegaly were easily detected. Since ultrasonography is noninvasive and can be repeated, ultrasonographic studies are useful for evaluating and monitoring abdominal complications which are frequently encountered in these transplant patients.

Sonographic assessment and grading of spleen size.

Ultrasonography was used to evaluate spleen size in patients with various clinical conditions including those of the liver, blood, collagen, or autoimmune disease. To express spleen size, a spleen index (SI), the product of the transverse diameter and its perpendicular diameter measured on the maximum cross-sectional image of the spleen, was used. SI correlated well with the volumes of resected spleens. Splenomegaly was present in high percentages of patients with liver, blood, collagen, and autoimmune disease, even though a majority of these spleens were not large enough to palpate. By grading the SI, characteristic distributions of SI were obtained for patients with different types of diseases. Obtaining and grading the SI by the use of ultrasound appears to be a significant supplemental aid for evaluating spleen size, especially in patients whose spleens are not palpable.

Thymosin ß4 Gene Expression in Leukemic Cells.

Thymosin ß4 (Tß4) was originally isolated as a thymic hormone from calf thymosin fraction 5 which exhibited both immune and endocrine functions in vivo and in vitro. Tß4 is a ubiquitous peptide located in various tissues of mammalian species and other vertebrate classes. Recent studies on the molecular cloning and sequence analysis of rat and human Tß4 cDNA have demonstrated that Tß4 lacks a signal peptide which makes it unlikely that Tß4 is a secretory peptide. The real function of Tß4 is presently unknown, however, we focus this review on the molecular biology of Tß4 with a special reference to the aspects of the Tß4 gene expression in leukemic cells and cell lines during growth and differentiation.

Expression Pattern of Mitochondrial ND2 Gene in Human Leukemia and in HL60 Cells During Growth and Differentiation.

We differentially screened 5,000 clones from a cDNA library of acute myelogenous leukemia (AML) cell line HL60 using cDNA probes derived from normal granulocytes or from acute myelomonocytic leukemia cells, the objective being to identify genes preferentially expressed in myeloid lineage leukemic cells. One clone, corresponding to a mitochondrial DNA fragment, including NADH dehydrogenase subunit 2 (ND2) gene, was selected for use as a probe. We examined expression of the ND2 gene in various leukemic cell populations and in normal peripheral blood cells. DNA-RNA hybridization studies revealed that ND2 messenger RNA (mRNA) was more markedly expressed in AML cells than in other leukemic cells and normal peripheral blood granulocytes. The expression of ND2 mRNA decreased in HL60 cells several hours after treatment with phorbol myristate acetate (PMA), or dimethyl sulfoxide (DMSO). However, the ND2 gene expression did not depend on the growth-state of HL60 cells because the steady-state level of its expression was observed during transitions of growth. These results suggest that ND2 mRNA is involved in the maturation of myeloid cells and in cellular differentiation, in a lineage-preferential manner. A comparison of the nucleotide sequence of this clone with the documented human mitochondrial DNA sequence revealed several single-base substitutions, insertions and a 39-bases insertion.

[Acute renal failure associated with acute type A hepatitis with a mild liver damage].

Acute renal failure associated with acute type A hepatitis was successfully treated with hemodialysis. Though acute renal failure is usually associated with severe liver damage of end stage cirrhosis or fulminant hepatitis, liver damage of our case was only slight. The etiology of the renal failure is discussed.

Expression of the thymosin beta 4 gene during differentiation of hematopoietic cells.

Thymosin beta 4 (T beta 4) was originally isolated as a thymic hormone. Its functional properties remain obscure; however, the N-terminal peptidic sequence could have a regulatory function on hematopoietic stem cell proliferation. To investigate the mechanism of T beta 4 expression, we studied T beta 4 gene expression in various leukemic cells and in established cell lines. Among leukemic cell samples obtained from leukemia patients, the T beta 4 gene was highly expressed in a lymphoid lineage, especially in adult T-cell leukemia (ATL) cells, rather than in a granulocyte lineage. The T beta 4 gene was more transcriptionally active in chronic B-cell leukemia than in acute B-cell leukemia, while it was inactive in plasma cell leukemia. We also found that cells from one of the ATL patients transcribed a heterogeneous message. T beta 4 messenger RNA increased in MOLT-3 during differentiation by 12-O-tetradecanoylphorbol-13-acetate (TPA), in HL60 cells induced by TPA or dimethylsulfoxide and K562 cells stimulated by cytosine arabinoside or hemin. The genomic sequence of T beta 4 is considered to be highly conserved. Only 1 of 20 genomes from normal or hematopoietic malignant cells showed restriction fragment length polymorphism. These findings, along with previous data, suggest that T beta 4 may be a new marker of differentiation of hematopoietic cells.

[Abrupt onset and rapid deterioration in the course of congenital ornithine transcarbamylase deficiency: a case report].

We report a 17-year-old female case of ornithine transcarbamylase (OTC) deficiency who died of brain edema due to hyperammonemic attack. The patient had a brother with OTC deficiency who had died of hyperammonemia at 17 years of age. She firstly had a symptom of headache, nausea, vomiting and myalgia at 14 years old and twice thereafter. On admission she had a severe disorientation and vomiting. The plasma ammonia level was 89 micrograms/dl, then increased to 400 micrograms/dl in five hours. In addition to plasma exchange, hemodialysis and then peritoneal dialysis for next 5 days, parenteral sodium benzoate and arginine were administered. Although the plasma ammonia level improved gradually, her consciousness never returned and she died of severe brain edema with uncontrollable hypotension on day 8. Histology of a necropsy liver sample showed fatty metamorphosis of hepatocytes mainly with fine lipid droplets. Electron micrograph of hepatocytes showed crystalloid inclusions in mitochondria. Significance of the clinical course and the treatment during hyperammonemic crisis was discussed.

[Giant hepatic cyst effectively treated by transdermal transhepatic ethanol injection therapy. A case report].

Giant hepatic cysts were effectively treated by transhepatic ethanol injection therapy. A 68-year-old male and a 70-year-old female were admitted to Kyushu University Hospital because of epigastric fullness. Ultrasound and CT examination disclosed a giant cyst in the left lobe of the liver of both patients. 99% ethanol of about a quarter of volume of the cysts were infused through catheter transdermal. After retaining for 15 minutes the ethanol was discarded and this procedure repeated twice. The cysts have been keeping a minimum size and the patients are free from symptoms for over one year.

Secretion of an interleukin 1-like activity by acute monocytic leukemia cells which inhibits growth of fibroblasts.

Acute monocytic leukemia cells (AMoL cells), obtained by leukapheresis, were cultured in vitro. In response to lipopolysaccharide, AMoL cells produced a large amount of thymocyte proliferation activity. The crude supernatants from AMoL cells inhibited fibroblast growth, in a dose-dependent manner. Upon gel filtration, the thymocyte proliferation activity had a molecular mass of 37,000 and 17,000 daltons, and was heat labile and fairly resistant to freezing and thawing. The fractions containing thymocyte proliferation activity additionally possessed an inhibitory activity for the growth of fibroblasts. These results suggest that AMoL cells may participate in the progress of the disease (leukemia), by secreting these cytokines.

[Sézary syndrome associated with hepatocellular carcinoma and monoclonal gammopathy--a case report].

We treated 54-year-old Japanese man with a large cell type of Sézary syndrome. He had generalized erythrodermia, superficial lymphadenopathy, atypical lymphocytes in the peripheral blood, anti-HTLV-I antibody negativity and chromosomal abnormality. The patient was a hepatitis B virus carrier, and was complicated with hepatocellular carcinoma and monoclonal gammopathy of IgG, lambda type. Sézary syndrome is a T cell malignancy, the clinical course of which is relatively mild and chronic; accordingly, this case showed no crisis under chemotherapy. However, the patient died due to rapid growth of the hepatoma. Although case reports of Sézary syndrome complicated with other malignancies are very few, the occurrence of malignancies is possible because of decreased immunological function in the patients. In this case, hepatitis B virus might participate in the hepatic oncogenesis under dysfunction of helper/inducer cells. In addition, the complication of monoclonal gammopathy was also interesting from the standpoint of the helper function of Sézary cells.

Human Ia-associated invariant chain gene has multiple transcription initiation sites in CLL cells.

We show a northern transfer experiment revealed two mRNA of Ia-associated invariant chain (In) gene in chronic lymphocytic leukemia (CLL) cells which are approximately 1580 and 1440 nucleotides in length. Primer extension experiment shows that less prominent transcript was found to initiate 140 nucleotides upstream from the major cap site. The newly identified cap site was preceded by CG rich sequence but no typical promotor sequence. Southern hybridization analysis with In cDNA probe indicates no recombination or amplification of In gene in the CLL cells. This is the first documented example of such a mode of expression in malignant cells in vivo.