Is the degree of fetal hepatosplenomegaly with transient abnormal myelopoiesis closely related to the postnatal severity of hematological abnormalities in Down syndrome?

We present three cases of transient abnormal myelopoiesis associated with trisomy 21 in which hepatomegaly was apparent during the fetal period. In the first case, the fetal hepatosplenomegaly was severe, multiple organ failure occurred in the neonatal period and death ensued at 4 weeks of age. In the second case, the hepatomegaly was moderate, and with conservative treatment in the neonatal period the outcome was good. In the third case, hepatomegaly was mild and improved spontaneously, and the hematological abnormalities required only conservative treatment in the neonatal period. Our experience raises the question of whether fetal hepatosplenomegaly is a predictor of transient myeloproliferative disorder with trisomy 21 and whether the degree of fetal hepatomegaly is a marker for the neonatal severity of hematological abnormalities.

Effects of calcium hydroxide on physical and sealing properties of canal sealers.

AIM: To investigate whether Ca(OH)2 in four different agents alters the physical properties (Exp. I) and sealing ability (Exp. II) of root canal sealers. EXPERIMENT: (Exp. I) Calcipex (Nippon Sika-Yakuhin, Shimonoseki, Japan), Vitapex (Neo-Dental, Tokyo, Japan), Calkyl (Showa Yakuhin, Tokyo, Japan), and Ca(OH)2 were used as Ca(OH)2 agents. Four sealers were tested for flow, working time, setting time, and film thickness: Canals (Showa Yakuhin), Canals-N (Showa Yakuhin), Ketac -Endo (Espe, Seefeld, Germany), and Sealapex (Kerr, Romulus, MI, USA). Each Ca(OH)2 agent was added to 10 vol.% of each sealer, and the mixture and controls without a Ca(OH)2 agent were tested according to ISO specifications. Measurements were compared using Student's t-tests (P < 0.05). (Exp. II) After removing Ca(OH)2 agents applied to the root canals of 100 extracted human teeth, canals were filled with sealer. Controls were filled with each sealer without Ca(OH)2 agents. Sealing ability was evaluated using distance of dye penetration from the apices. Dye penetration data were compared using analysis of variance and post hoc Newman-Keuls test (P < 0.05). RESULTS: Ca(OH)2 agents influenced the physical properties of the sealers. Flow and setting time met ISO requirements, but film thickness and working time did not. Apical sealing ability of all four sealers was influenced by Ca(OH)2 agents. The sealing ability of Sealapex improved with all Ca(OH)2 agents. The physical and sealing abilities varied among the other sealers. CONCLUSIONS: Contact with Ca(OH)2 agents left on the canal wall caused considerable changes to the sealing ability of sealers.

Changes in enamel surface roughness and adhesion of Streptococcus mutans to enamel after vital bleaching.

OBJECTIVES: The purpose of this in vitro study was to observe the influence of vital bleaching on changes to the enamel surface and adhesion of Streptococcus mutans to tooth enamel. METHODS: The coronal part of each of 70 extracted third molars was cut in half, with either the buccal or lingual half used for experiments or controls. Experimental halves were assigned to the following conditions: (A) enamel was bleached 1, 3 or 5 times using a bleaching material with or without etching; or (B) etched condition without bleaching. All control samples were kept intact in physiological saline solution. Surface roughness (Ra; roughness center-line average: microm) of enamel was measured for 35 pairs of specimens. TS broth culture medium containing 3% glucose was inoculated with S. mutans and cultured for 72 h before adding the other 35 pairs of specimens. Under scanning electron microscopy, the number of S. mutans colonies was counted and statistically analysed. RESULTS: Compared to controls, bleached enamel displayed increased colonies of S. mutans. Repeated bleaching further increased bacterial adhesion and maximal colonies counts were found under conditions of five bleaching treatments plus etching (p

Identification of a novel fusion gene, TTL, fused to ETV6 in acute lymphoblastic leukemia with t(12;13)(p13;q14), and its implication in leukemogenesis.

ETS variant gene 6 (ETV6)/translocation, ETS, leukemia (TEL)-involving chromosomal translocations are frequently observed in various hematologic neoplasms. We describe here a novel ETV6-involving translocation, t(12;13)(p13;q14), found in the case of acute lymphoblastic leukemia, in which ETV6 fused with a previously unknown gene, named Twelve-thirteen Translocation Leukemia gene (TTL), at 13q14. TTL was weakly but ubiquitously expressed in normal human tissues as detected by reverse transcribed-PCR. Three TTL splicing forms were identified, TTL-T from a human testis cDNA library, with an open-reading frame of 402 bp encoding 133 amino acids (aa), and TTL-B1 and -B2 from a human brain cDNA library. These proteins have no homology to known proteins. In leukemic cells from the patient, both reciprocal fusion transcripts, ETV6/TTL and TTL/ETV6, were expressed. The predominant fusion transcript, TTL/ETV6-1, encodes a predicted 530 aa fusion protein containing 89 aa of the N-terminal TTL fusing to the helix-loop-helix domain and ETS-binding domain of ETV6. Although the function of TTL is yet to be elucidated, our findings will provide another insight into the molecular pathogenesis of leukemia having ETV6-involving translocations.

Resting energy expenditure among Japanese.

1. Resting energy expenditure (REE) provides appropriate basic data for the calculation of energy requirements. 2. The REE of 6498 subjects according to sex and age (1 year stratification), with a minimum of 10 subjects per group, was measured systematically using the easy portable calorimeter (Metavine; Vine, Tokyo, Japan). 3. The REE or the REE/kg according to age and sex was observed to obtain the amount of standard deviation (20-25%). 4. The REE/kg for male and female subjects was maintained at a steady level after the age of 15 years and was estimated to be around 29 kcal/kg.

Intubating laryngeal mask airway size selection: a randomized triple crossover study in paralyzed, anesthetized male and female adult patients.

UNLABELLED: We determined the optimal size of intubating laryngeal mask airway (ILM) for ventilation and blind tracheal intubation in men and women. We also determined the distance the tracheal tube needs to protrude beyond the distal aperture to ensure that the cuff is through the vocal cords. Fifty male and 50 female anesthetized, paralyzed patients (ASA physical status I or II, aged 18-80 yr) were studied. Three operators (A, B, and C) were involved for the purposes of blinding. The size 3, 4, or 5 ILM was inserted into each patient in random order by Operator A, and the quality of ventilation was scored (adequate, suboptimal, or failed) by Operator B. The fiberoptic position (correct, too shallow, or too deep) and the distance between the distal aperture and the vocal cords was determined by Operator B. A single attempt at blind intubation was made by Operator C. Operators B and C were blinded to the size of the ILM. Operator C was also blinded to the information recorded by Operator B. All ILMs were inserted into the laryngopharynx at the first attempt. For men and women, the ventilation score was smaller for the Size 3 than the Size 4 or 5 (all: P < 0.002). For men, correct positioning was less common with the Size 3 than the Size 4 or 5 (both: P < 0.02). For women, correct positioning was similar among sizes. For men, tracheal intubation was successful less frequently with the Size 3 (84%) than the Size 4 (100%) or 5 (98%) (both: P < or = 0.01). For women, tracheal intubation success was similar among sizes (Size 3, 4, and 5: 86%, 96%, and 92%, respectively). Intubation was always successful if the ILM was correctly positioned and always failed if it was too shallow or deep. In both male and female patients, the distance between the distal aperture and the vocal cords increased with increasing ILM size (all: P < 0.04) and patient height (P < 0.0001) and was always longer for men (all: P < 0.0001). The overall mean distance (95% confidence interval) that the tracheal tube needed to protrude was 10-12 cm (8-13 cm) in men and 8-11 cm (8-12 cm) in women. We conclude that for men, the Size 4 and 5 ILMs are better than the Size 3 for ventilation and blind intubation. For women, the Size 4 and 5 ILMs are better than the Size 3 for ventilation, but there is no difference among sizes for blind intubation. The length the tracheal tube must protrude from the distal aperture to ensure that the cuff is completely through the vocal cords is 8-13 cm, depending on ILM size, the tracheal tube size, and the sex and height of the patient. IMPLICATIONS: For men, the Size 4 and 5 intubating laryngeal mask airways are better than the Size 3 for ventilation and blind tracheal intubation. For women, the Size 4 and 5 are better than the Size 3 for ventilation, but there is no difference among sizes for blind intubation. The length the tracheal tube must protrude from the distal aperture of the intubating laryngeal mask airway to ensure that the cuff is completely through the vocal cords is 8-13 cm, depending on the size of the mask and tracheal tube and on the sex and height of the patient.

Resting energy expenditure among Japanese.

1. Resting energy expenditure (REE) provides appropriate basic data for the calculation of energy requirements. 2. The REE of 6498 subjects according to sex and age (1 year stratification), with a minimum of 10 subjects per group, was measured systematically using the easy portable calorimeter (Metavine; Vine, Tokyo, Japan). 3. The REE or the REE/kg according to age and sex was observed to obtain the amount of standard deviation (20-25%). 4. The REE/kg for male and female subjects was maintained at a steady level after the age of 15 years and was estimated to be around 29 kcal/kg.

Notch1 inhibits differentiation of hematopoietic cells by sustaining GATA-2 expression.

Notch signaling is involved in cell fate decisions in many systems including hematopoiesis. It has been shown that expression of an activated form of Notch1 (aNotch1) in 32D mouse myeloid progenitor cells inhibits the granulocytic differentiation induced by granulocyte colony-stimulating factor (G-CSF). Results of the current study show that aNotch1, when expressed in F5-5 mouse erythroleukemia cells, also inhibits erythroid differentiation. Comparison of the expression levels of several transcription factors after stimulation for myeloid and erythroid differentiation, in the presence or absence of aNotch1, revealed that aNotch1 did not change its regulation pattern with any of the transcription factors examined, except for GATA-2, despite its inhibitory effect on differentiation. GATA-2 was down-regulated when the parental 32D and F5-5 were induced to differentiate into granulocytic and erythroid lineages, respectively. In these induction procedures, however, the level of GATA-2 expression was sustained when aNotch1 was expressed. To ascertain whether maintenance of GATA-2 is required for the Notch-induced inhibition of differentiation, the dominant-negative form of GATA-3 (DN-GATA), which acted also against GATA-2, or transcription factor PU.1, which was recently shown to be the repressor of GATA-2, was introduced into aNotch1-expressing 32D (32D/aNotch1) cells that do not express GATA family proteins other than GATA2. Both DN-GATA and PU.1 reversed the phenotype of 32D/aNotch1 inducing its differentiation when G-CSF was added. Furthermore, enforced expression of HES-1, which is involved in Notch signaling, delayed differentiation of 32D, and again this phenotype was neutralized by DN-GATA. These results indicate that GATA-2 activity is necessary for the Notch signaling in hematopoietic cells.

Calcium concentration and pH of the periapical environment after applying calcium hydroxide into root canals in vitro.

The purpose of the present study was to determine both calcium concentration and pH in the periapical region after application of 1 of 4 different calcium hydroxide preparations into experimental root canals. Fifty root canal models were divided into five groups: group 1--calcium hydroxide was mixed with distilled water at a powder/water weight ratio of 38%; group 2--calcium hydroxide was mixed with distilled water at 44%; group 3--calcium hydroxide was mixed with distilled water at 50%; group 4--calcium hydroxide powder only was used; and group 5-the control group, in which nothing was applied to the canals. All samples were immersed in distilled water maintained at 37 degrees C. Calcium concentration and pH of the distilled water were measured after 3 days, 7 days, and then at weekly intervals up to 15 wk, during which time the storage medium was renewed after each measurement. Calcium concentration and the change in pH of the distilled water were statistically quicker and greater in groups 1 to 3 (mixture groups) than group 4 (powder only) (p < 0.05). The highest calcium concentration (peak Ca2+ release) was observed after 3 days for the mixture groups, whereas that for the powder only group was found at 7 days. Peak pH change was found after 14 days for the mixture groups, whereas that for the powder only group was found at 49 days. After peaking, all groups showed a decline of the pH over time. These results suggest that the time required for optimum intracanal activity when using calcium hydroxide mixtures is at least 2 wk.

Changes in molecular weight of hyaluronan and hyaluronidase activity in uterine cervical mucus in cervical ripening.

BACKGROUND: This study was undertaken to investigate changes in the forms of hyaluronan and hyaluronidase activity in cervical mucus during cervical ripening. METHODS: Uterine cervical mucus was obtained from 57 pregnant women (25 at preterm gestation, ten at term gestation, 11 within 1 week before labor, and 11 during the first stage of labor). We determined 1) concentration of hyaluronan, 2) hyaluronidase activity, and 3) molecular weight of hyaluronan in cervical mucus. Data are presented as mean and range. RESULTS: The hyaluronan concentration in mucus in the 1st stage of labor (1.58 microg/ml, 0.46-23.96) was significantly (p<0.05) higher than that in all other groups (preterm: 0.29, 0.10-0.88; term: 0.24, 0.11-0.80; within 1 week before labor: 0.30, 0.18-0.62). Hyaluronidase activity both within 1 week before labor group (3.03 min., 1.12-3.95) and in 1st stage of labor group (3.52, 0.43-5.15) was significantly (p<0.05) higher than that in preterm group (1.70, 0.00-5.47). The molecular weight of hyaluronan in cervical mucus in the 1st stage of labor (0.97x106, 0.86-1.41) was significantly (p<0.05) lower than in the preterm and term groups (preterm: 1.60, 1.21-2.20, term: 1.41, 1.21-2.20). There was a significant correlation between hyaluronidase activity and molecular weight of hyaluronan (p<0.05, r=-0.41, n=23). CONCLUSION: These findings suggest that either hyaluronidase or low-molecular weight hyaluronan could be one of the most important regulators in the process of cervical ripening.

The influence of two concentrations of sodium hypochlorite on human blood: changes in haemolysis, pH and protein.

AIM: To evaluate the in vitro effect of 2.5% and 5.0% sodium hypochlorite (NaOCl) on human blood. METHODOLOGY: Each concentration of NaOCl was reacted with human blood for 5 min at volume ratios of 1 : 1, 1 : 6, 1 : 12, each creating changes in colour, pH and temperature. Reaction suspensions were separated by centrifugation, and absorption measurements made for separated bilirubin, Fe, and protein supernatants. Each supernatant was desalted, lyophilized, and treated by SDS polyacrylamide gel electrophoresis (SOS-PAGE). RESULTS: Increased ratios and concentrations of NaOCl caused an increase in both pH and temperature. Protein supernatants tended to decompose on SDS-PAGE. Supernatants showed increased decolourisation with 5.0% NaOCl. Concentrations of bilirubin, Ferrum and protein in supernatants decreased with increased NaOCl concentration. NaOCl had an effect on the protein component in blood. CONCLUSIONS: These data suggest that changes in molecular structure are due to the chemical effects of NaOCl. Protein bands tended to show low molecular weight, suggesting that haemoglobin components effect the oxidation-reduction reaction.

Vitamin E modified cellulose membrane.

There has been a demand for hemodialysis membranes of better biocompatibility, the use of which would reduce the incidence of complications in patients who have been under long hemodialysis treatment. We developed a surface modification technique consisting of forming efficiently a synthetic polymer layer on the inside of the regenerated cellulose hollow fiber without impairing fiber performance. Our newly developed membrane has excellent biocompatibility by modifying the inner surface and by immobilizing vitamin E (alpha-tocopherol), which serves as an antioxidant, to the modified surface.

Relationship between mutations in dihydropteroate synthase of Pneumocystis carinii f. sp. hominis isolates in Japan and resistance to sulfonamide therapy.

We examined mutations in the dihydropteroate synthase (DHPS) genes of Pneumocystis carinii f. sp. hominis (P. carinii) strains isolated from 24 patients with P. carinii pneumonia (PCP) in Japan. DHPS mutations were identified at amino acid positions 55 and/or 57 in isolates from 6 (25.0%) of 24 patients. The underlying diseases for these six patients were human immunodeficiency virus type 1 infection (n = 4) or malignant lymphoma (n = 2). This frequency was almost the same as those reported in Denmark and the United States. None of the six patients whose isolates had DHPS mutations were recently exposed to sulfa drugs before they developed the current episode of PCP, suggesting that DHPS mutations not only are selected by the pressure of sulfa agents but may be incidentally acquired. Co-trimoxazole treatment failed more frequently in patients whose isolates had DHPS mutations than in those whose isolates had wild-type DHPS (n = 4 [100%] versus n = 2 [11.1%]; P = 0.002). Our results thus suggest that DHPS mutations may contribute to failures of co-trimoxazole treatment for PCP.

Genotyping of Pneumocystis carinii f. sp. hominis isolates in Japan based on nucleotide sequence variations in internal transcribed spacer regions of rRNA genes.

Genotyping of Pneumocystis carinii (Pc) isolated from 24 bronchoalveolar lavage (BAL) fluid specimens in Japan was examined based on nucleotide sequence variations in internal transcribed spacer regions 1 and 2 (ITS1 and ITS2, respectively) of rRNA genes. We found 11 ITS1 genotypes including 2 novel ones and 11 ITS2 genotypes including 3 new ones. Combining the ITS1 and ITS2 genotypes resulted in 30 ITS genotypes, of which 10 are newly described in this report. Two or more genotypes in ITS regions in a specimen were observed in 16 of 24 patients. Our results will be of help for the epidemiological investigation of Pc infection.

Binding of Delta1, Jagged1, and Jagged2 to Notch2 rapidly induces cleavage, nuclear translocation, and hyperphosphorylation of Notch2.

Delta1, Jagged1, and Jagged2, commonly designated Delta/Serrate/LAG-2 (DSL) proteins, are known to be ligands for Notch1. However, it has been less understood whether they are ligands for Notch receptors other than Notch1. Meanwhile, ligand-induced cleavage and nuclear translocation of the Notch protein are considered to be fundamental for Notch signaling, yet direct observation of the behavior of the Notch molecule after ligand binding, including cleavage and nuclear translocation, has been lacking. In this report, we investigated these issues for Notch2. All of the three DSL proteins bound to endogenous Notch2 on the surface of BaF3 cells, although characteristics of Jagged2 for binding to Notch2 apparently differed from that of Delta1 and Jagged1. After binding, the three DSL proteins induced cleavage of the membrane-spanning subunit of Notch2 (Notch2(TM)), which occurred within 15 min. In a simultaneous time course, the cleaved fragment of Notch2(TM) was translocated into the nucleus. Interestingly, the cleaved Notch2 fragment was hyperphosphorylated also in a time-dependent manner. Finally, binding of DSL proteins to Notch2 also activated the transcription of reporter genes driven by the RBP-Jkappa-responsive promoter. Together, these data indicate that all of these DSL proteins function as ligands for Notch2. Moreover, the findings of rapid cleavage, nuclear translocation, and phosphorylation of Notch2 after ligand binding facilitate the understanding of the Notch signaling.

Effect of canal drying methods on the apical seal.

The purpose of this study was to observe the seal obtained in human root canals under different in vitro treatment conditions. Root canals of freshly extracted human maxillary incisors received traditional biomechanical preparation and irrigation with 10% NaClO, followed by a final rinse of distilled water. Teeth were divided into four groups. In group 1, each root canal was dried with one paper point. In group 2, each root canal was dried with four paper points. Group 3 was dried with four paper points, followed by a gentle burst of warm air. Group 4 was dried using four paper points, followed by an internal 200 degrees C heat probe. Twenty canals in each group were filled with a gutta-percha cone and a zinc oxide-eugenol sealer, whereas 20 were filled with a glass ionomer sealer. All teeth were immediately immersed in India ink for 1 or 4 wk. The teeth were cleared, and the dye penetration-leakage measured with an accuracy of +/- 0.01 mm. Optimum sealing conditions were observed when the canal was dried with paper points and a 200 degrees C probe. There were no significant differences between 1 and 4 wk immersion. Glass ionomer sealer appeared more susceptible to the wet condition of the root canal walls than zinc oxide-eugenol sealer. An additional 40 teeth were prepared in the same manner as the dye penetration tests to observe the drying conditions of the root canal walls, and the moisture inside the canals was measured to an accuracy of 0.0001 g. The highest degree of internal canal wall dryness was found in group 4.

The walking bleach procedure: an in vitro study to measure microleakage of five temporary sealing agents.

The purpose of this study was to compare the in vitro sealing capacity of five materials, each used as a temporary sealing agent for the walking bleach technique. All teeth received traditional biomechanical root canal instrumentation, after which the walking bleach agent was placed in the pulp chamber space. The occlusal access was sealed with one of five temporary materials: two hydraulic filling materials, a photoactivated resin composite, a zinc oxide-eugenol cement, and a zinc oxide phosphate cement with/without the placement of a piece of rubber sheet that was placed as a barrier to isolate filling material from the bleaching agent. All teeth were stored in a 1% solution of Alcian blue with thermal cycling stress. After 1 wk, they were sectioned longitudinally, and ranked by graded scores of 0 to 3, according to the degree of the dye penetration. Significantly less dye microleakage was observed in the two hydraulic materials than in the photoactivated resin. Both zinc oxide-eugenol and zinc phosphate cements showed a considerable amount of microleakage. There were no significant differences between the groups with and without a rubber sheet. Our data demonstrate that hydraulic filling materials provide the most favorable cavosurface seal when they are firmly packed into the cavity space to prevent microleakage.

Mouse jagged1 physically interacts with notch2 and other notch receptors. Assessment by quantitative methods.

The Delta/Serrate/LAG-2 (DSL) domain containing proteins are considered to be ligands for Notch receptors. However, the physical interaction between DSL proteins and Notch receptors is poorly understood. In this study, we cloned a cDNA for mouse Jagged1 (mJagged1). To identify the receptor interacting with mJagged1 and to gain insight into its binding characteristics, we established two experimental systems using fusion proteins comprising various extracellular parts of mJagged1, a "cell" binding assay and a "solid-phase" binding assay. mJagged1 physically bound to mouse Notch2 (mNotch2) on the cell surface and to a purified extracellular portion of mNotch2, respectively, in a Ca(2+)-dependent manner. Scatchard analysis of mJagged1 binding to BaF3 cells and to the soluble Notch2 protein demonstrated dissociation constants of 0.4 and 0.7 nM, respectively, and that the number of mJagged1-binding sites on BaF3 is 5,548 per cell. Furthermore, deletion mutant analyses showed that the DSL domain of mJagged1 is a minimal binding unit and is indispensable for binding to mNotch2. The epidermal growth factor-like repeats of mJagged1 modulate the affinity of the interaction, with the first and second repeats playing a major role. Finally, solid-phase binding assay showed that Jagged1 binds to Notch1 and Notch3 in addition to Notch2, suggesting that mJagged1 is a ligand for multiple Notch receptors.

Mutation analysis of the WT1 gene in myelodysplastic syndromes.

The WT1 tumor suppressor gene was examined for mutations in a panel of 44 patients with myelodysplastic syndromes (MDS) including acute myelogenous leukemias (AML) secondary to MDS, using polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) analysis and sequencing analysis. A WT1 mutation was detected in one out of 17 cases of AML secondary to MDS. This mutation exists upstream of the zinc finger region and is predicted to produce a truncated WT1 protein lacking the zinc finger region. No mutations were detected in 27 MDS patients who had not progressed to AML. This is the first report of analysis for WT1 mutations in a large number of MDS patients, suggesting that WT1 mutations are uncommon in MDS. Abnormalities in this gene may, however, contribute to a small proportion of cases showing progression from MDS into AML.