[Coagulation and fibrinolysis system in pediatric cardiopulmonary bypass].

Coagulation and fibrinolysis system was evaluated during and after pediatric cardiopulmonary bypass (CPB. Twenty-two atrial septal defect (ASD) patients were surgically repaired under CPB and aortic cross-clamp through right thoracotomy. Drainage was established by gravity, CPB flow was kept 2.4 l/min/m2 and ACT was controlled over 400 seconds. HCT, PLT, fibrinogen, AT-III, D-dimer, thrombin-antithrombin complex (TAT), alpha2 plasmin inhibitor-plasmin complex (PIC), and plasminogen activator inhibitor (PAI-1) were measured at 6 points [after induction of anesthesia, 10 minutes after initiating CPB, end of CPB, on the entrance of intensive care unit (ICU), postoperative day (POD) 1, and at outpatient division]. Both fibrinogen and AT-III showed low values during CPB (121.9 +/- 22.0 mg/dl, 57.6 +/- 10.6%). D-dimer increased at 1 week postoperatively in all patients (5.57 +/- 3.45 microg/ml). There were significantly positive correlations between CPB duration and TAT value at the end of CPB (r = 0.88, p < 0.01), on the entrance of ICU (r = 0.71, p < 0.01). There was also a positive correlation between CPB duration and PIC value on the entrance of ICU (r = 0.53, p < 0.01). Five patients showed high PAI-1 value on the entrance of ICU, which remained high in 2 of them on POD 1. The outcomes from the current study suggest that there is a potential of coagulation-dominant disseminated intravascular coagulation (DIC) during pediatric CPB even in ASD patients who do not need long CPB. Longer CPB and severe hemodilution might become risk factors.

A comparative study of immunological and cytochemical profiles between adult and childhood acute lymphoblastic leukemias (ALLs): heterogeneity in adult common ALL.

To investigate the biological differences between adult and childhood acute lymphoblastic leukemia (ALL), leukemic blasts from 33 patients with ALL (22 adults and 11 children) and from 11 patients in the lymphoid crisis of chronic myeloid leukemia (CML) were studied using cytochemical and immunological markers and also by the outcome of their treatment. The cytochemical studies showed that blasts from seven of the adult ALL patients were dense-granular-positive (DG-positive) for beta-glucuronidase, whereas the blasts from the children were negative except for one (with T-ALL). In the adults with common ALL (cALL), survival of patients DG-positive for this enzyme were significantly shorter than that of eight patients with a scattered granular pattern (p less than 0.05). The mean ratio between the percentage of blasts positive for cALL antigen (cALLA) to that of blasts positive for terminal deoxynucleotidyl transferase (TdT) in the adult group with cALL (0.6 +/- 0.3) was significantly lower (p less than 0.01) than in the group of children with cALL (1.1 +/- 0.2) or in the lymphoid-crisis group (1.5 +/- 1.0). These findings indicate that adult cALL consists of two distinct subpopulations, one with less differentiated phenotype (cALL-/TdT+) and the other with more (cALL+/TdT+). In contrast, the blast cells in childhood cALL and some patients in lymphoid crisis had a relatively homogeneous population with the latter phenotypes. The results suggest that the clonotypic cells in adult ALL, particularly in cALL, appear to be more immature than those in childhood ALL. The beta-glucuronidase patterns indicate a further heterogeneity in adult ALL.

Analysis of growth failure in children treated for acute lymphoblastic leukemia.

Growth patterns were surveyed in 303 children with acute lymphoblastic leukemia who had remained in continuous first complete remission for a minimum of 1 year (median 3 years). Chemotherapy was given for 3 years, and central nervous system prophylaxis consisted of cranial irradiation at a total dose of 18 Gy or intravenous high-dose methotrexate (1.5-6 g/m2 for three to six doses). Intensive chemotherapy, including cyclophosphamide, doxorubicin, and cytosine arabinoside was given to children with high-risk features. Two of 205 children with low- or intermediate-risk features and 13 of 98 children with high-risk features showed a decrease in the growth rate of less than -2 SD. In 14 of these 15 patients, the age at onset was over 9 years and growth failure became most predominant in the prepubertal period: ten of these children showed a tendency toward delayed pubertal development, but eight showed later catch-up growth with pubertal maturation after completion of chemotherapy. Thus, chemotherapy appeared to contribute temporarily to the growth failure and gonadal impairment that occurred in the prepubertal period. No obvious correlation between the administered cranial irradiation and growth failure was found, but further study with a longer follow-up will be necessary to determine the long-term effects of irradiation on subsequent growth patterns in children.

[Experimental studies on hepatic circulation: effects of drugs on blood flows in liver tissue and portal vein of the cats].

The effects of some drugs on the hepatic circulation were examined by thermoelectrical and electromagnetic methods under pentobarbital-anesthesia in normal and CCl4-pretreated cats. The following results were obtained. 1. Both adrenergic alpha and beta receptor functions were involved in the regulation of the hepatic circulation of normal cats. 2. Three calcium blockers (nifedipine, nicardipine, diltiazem) had different potencies in increasing the hepatic blood flow of normal cats. 3. The isolated veins including portal vein showed the regional difference in the responsiveness to calcium blockers. 4. In CCl4-pretreated cats, adrenergic alpha receptor function was dominant in the control of hepatic circulation and diltiazem raised portal venous pressure.

The soybean GH2/4 gene that encodes a glutathione S-transferase has a promoter that is activated by a wide range of chemical agents.

Transcriptional activation of the soybean (Glycine max) GH2/4 gene (also referred to as Gmhsp26-A) and increase in abundance of the GH2/4 mRNA (also referred to as pCE54) have been previously shown to occur following treatment of soybean seedlings with auxins, nonauxin analogs, heavy metals, and a variety of other agents. To determine whether the GH2/4 promoter is responsive to an array of different agents, we have analyzed the inducibility of the GH2/4 promoter fused to the beta-glucuronidase reporter gene in transgenic tobacco (Nicotiana tabacum) plants. We have shown that a wide variety of chemical agents induce this promoter in a tissue-specific and concentration-dependent manner. In addition, we have used an affinity-purified antibody raised against recombinant GH2/4 protein to show that the GH2/4 protein increases in response to auxin application and is localized in the cytosol of soybean cells. Recombinant GH2/4 protein can be purified to homogeneity on a glutathione-agarose resin, and the purified protein has glutathione S-transferase activity when assayed with the substrate 1-chloro-2,4-dinitrobenzene.

Cloning of genes encoding auxin-binding proteins (ABP19/20) from peach: significant peptide sequence similarity with germin-like proteins.

An auxin-binding protein (ABP) was previously isolated from shoot apices of peach trees to homogenity on standard SDS-PAGE. Analysis of low-bis SDS-PAGE and direct peptide sequencing of purified peach ABP demonstrated that the ABP was composed of two types of polypeptides (designated ABP19 and ABP20). Several cDNA and genomic clones which encode peach ABPs were obtained and analysed. We found that there are at least three classes of ABPs in the peach genome. Open reading frames of these ABPs were 627 bp, predicting a 209 amino acid polypeptide of 22 kDa. An N-terminal hydrophobic signal sequence of 18 amino acids and a putative N-glycosylation site at N60-T-T/S were deduced. Homology search analysis revealed that ABP19 is highly homologous to proteins related to the germin family. The deduced amino acid sequence of ABP19 showed very low overall sequence homology with ABP1, an ABP isolated from maize coleoptile, but it contained a small region which shared 40% homology with a putative auxin binding site in ABP1 (BoxA). In addition, the sequence surrounding the region is highly conserved among peach ABPs and the germin family.

Isolation and characterization of a 60 kDa 2,4-D-binding protein from the shoot apices of peach trees (Prunus persica L.); it is a homologue of protein disulfide isomerase.

To obtain a candidate auxin-binding protein (ABP), a soluble 60 kDa protein was isolated from an extract of shoot apices of peach trees (Prunus persica L.) by affinity chromatography on a 2,4-dichlorophenoxyacetic acid (2,4-D)-linked Sepharose4B column. The 60 kDa polypeptide, designated Pp60, was purified as a single band on SDS-PAGE by column chromatography. Its dissociation constant (Kd) for [14C]-2,4-D was calculated to be 3.5 x 10(-5) M. The binding of Pp60 for [14C]-2,4-D was inhibited by naphthalene-1-acetic acid (NAA) and p-chlorophenoxyisobutyric acid (PCIB) as well as 2,4-D. Indole-3-acetic acid (IAA) had little effect on the binding. These results suggested that Pp60 is a protein that has an affinity for 2,4-D, NAA, and PCIB in vitro. The partial amino acid sequences of Pp60 showed high homology to those of protein disulfide isomerase (EC 5.3.4.1). Immunoblot analysis demonstrated that Pp60 exists ubiquitously in shoots and leaves. In fruit, expression of Pp60 is restricted at an early stage of development.

Structural changes in the plastid DNA of rice (Oryza sativa L.) during tissue culture.

To investigate the rearrangement of the plastid genome during tissue culture, DNA from rice callus lines, which had been derived individually from single protoplasts isolated from seed or pollen callus (protoclones), was analyzed by Southern hybridization with rice chloroplast DNA (ctDNA) clones as probes. Among 44 long-term cultured protoclones, maintained for 4, 8 or 11 years, 28 contained plastid DNA (ptDNA) from which portions had been deleted. The ptDNA of all protoclones that had been maintained for 11 years had a deletion that covered a large region of the plastid genome. The deletions could be classified into 15 types from their respective sizes and positions. By contrast, no deletions were found in the ptDNA of 38 protoclones that had been maintained for only 1 month. These results indicate that long-term culture causes deletions in the plastid genome. Detailed hybridization experiments revealed that plastid genomes with deletions in several protoclones were organized as head-to-head or tail-to-tail structures. Furthermore, ptDNAs retained during long-term culture all had a common terminus at one end, where extensive rearrangement is known to have occurred during the speciation of rice and tobacco. Morphological analysis revealed the accumulation of starch granules in plastids and amyloplasts in protoclones in which the plastid genome had undergone deletion. Our observations indicated that novel structural changes in the plastid genome and morphological changes in the plastid had occurred in rice cells during long-term tissue culture. Moreover, the morphological changes in plastids were associated with deletions in the plastid genome.