Impact of mother donor, peripheral blood stem cells and measurable residual disease on outcomes after haploidentical hematopoietic cell transplantation with post-transplant cyclophosphamide in children with acute leukaemia.

Haploidentical hematopoietic-cell transplantation using post-transplant cyclophosphamide(Haplo-PTCy) is a feasible procedure in children with haematologic malignancies. However, data of a large series of children with acute leukaemia(AL) in this setting is missing. We analysed 144 AL Haplo-PTCy paediatric recipients; median age was 10 years. Patients had acute lymphoblastic(ALL; n = 86) or myeloblastic leukaemia(AML; n = 58) and were transplanted in remission(CR1: n = 40; CR2: n = 57; CR3+: n = 27) or relapse (n = 20). Bone marrow was the graft source in 57%; donors were father (54%), mother (35%), or sibling (11%). Myeloablative conditioning was used in 87%. Median follow-up was 31 months. At day +100, cumulative incidence (CI) of neutrophil recovery and acute GVHD (II-IV) were 94% and 40%, respectively. At 2-years, CI of chronic GVHD and relapse, were 31%, 40%, and estimated 2-year overall survival (OS), leukaemia-free survival (LFS) and graft-versus-host-relapse-free survival (GRFS) were 52%, 44% and 34% respectively. For patients transplanted in remission, positive measurable residual disease (MRD) prior to transplant was associated with decreased LFS (p = 0.05) and GRFS (p = 0.003) and increased risk of relapse (p = 0.02). Mother donor was associated with increased risk of chronic GVHD (p = 0.001), decreased OS (p = 0.03) and GRFS (p = 0.004). Use of PBSC was associated with increased risk of chronic GVHD (p = 0.04). In conclusion, achieving MRD negativity pre-transplant, avoiding use of mother donors and PBSC as graft source may improve outcomes of Haplo-PTCy in children with AL.

Enhanced expression of cellular prion protein gene by insulin or nerve growth factor in immortalized mouse neuronal precursor cell lines.

In order to understand the fundamental and putative roles of PrP(c) in the central nervous system, neuronal cell lines were established. Cells were immortalized by recombinant retrovirus vector-mediated transduction of SV40 T-antigen gene. Among these, two cell lines were selected based on their RT-PCR expressions of neuron-specific neurofilament (NF-H, NF-M) and cell morphology. These cell lines showed the properties of neuronal progenitor cells in antigenicity, morphology and responses to differentiating agents. Expression of PrP(c) was detected by immunocytochemical analysis. These cell lines responded to differentiating agents such as dibutyl cyclic AMP (dcAMP) and phorbol 12-myristate 13-acetate (PMA) before developing into neuronal-like cells. Neurite extensions were observed 20 min after incubation with the differentiating agents. Treatment with nerve growth factor (NGF) and insulin induced cell differentiation and enhanced expression of PrP gene (Prnp) mRNA and protein. The latter phenomenon was not inhibited by wortmannin, which is a specific inhibitor of phosphatidylinositol 3-kinase. These results suggest that PrP(c) plays an important role in the differentiation-mediated classic signaling pathway of neuronal cell.

[Influence of alcohol ingestion on plasma gastrin and CCK levels in human beings].

We investigated the time courses of the plasma ethanol, acetaldehyde, gastrin and cholecystokinin (CCK) levels after the ingestion of ethanol (1g/kg, 21.5%, whisky) in healthy male adult volunteers. The ethanol level reached a peak at 15 to 45 minutes after the ingestion and then decreased almost linearly. The acetaldehyde level of the group who became flushed after drinking (the F group) peaked earlier than that of the other group whose faces became only slightly flushed (the N group). The gastrin level increased significantly and remained elevated for about 3 hours. In three of the nine subjects, the CCK level increased 75 minutes after drinking. As a result, the absorption of the ingested ethanol is more rapid than 15 minutes, followed by a quick catabolism of the absorbed ethanol to acetaldehyde, but the catabolic action of acetaldehyde in the F group is later than that in the N group. The plasma gastrin level is certified to increase after the ingestion of ethanol and this is not mediated by acetaldehyde because the intra-venous infusion of acetaldehyde was not increased the plasma gastrin level in dogs. CCK which stimulate the pancreatic enzymes may be considered to have a possibility relating to the development of alcoholic acute pancreatitis but the ingestion of ethanol showed no increase of the plasma CCK level. But we cannot deny the possibility of the individual differences in the hormonal reaction in these experiments.

Teratogenic interaction of ethanol and hyperthermia in mice.

Alcohol and maternal hyperthermia have been implicated in human birth defects. Both ethanol and heat can induce neural tube defects (NTDs) and other developmental abnormalities in mice when large doses are given during pregnancy. To explore the teratogenic interaction of both agents, pregnant ICR mice were injected with a single dose of 25% ethanol and/or were heat-stressed in a water bath at 42 degrees C on the morning of Day 8 of gestation. Combined treatment with ethanol (0.01-0.02 ml/g) and heat (10 min), when they were given concurrently or 1 hr apart, resulted in a significant increase of resorptions and externally malformed fetuses. Skeletal malformations and visceral variations also increased significantly following a concurrent exposure to both agents. These results indicate that ethanol and heat can be synergistically teratogenic in mice when the doses of each agent are below the teratogenic threshold. It was also suggested that pretreatment with a small dose of ethanol may not enhance the teratogenicity of heat when the hyperthermic stress is strong enough and teratogenic by itself.

[Ultrasound-guided renal cyst puncture and 95% ethanol injection. Part 1: Estimation of ethanol levels in the blood and urine following 95% ethanol injection].

Seventeen solitary renal cysts were punctured with the ultrasound-guided procedure and 26-91% of the cyst volume was replaced with 95% ethanol used as a sclerosing agent of the cyst wall. Ethanol was injected through a 7 F pigtail ureteral catheter, allowed to remain in place for 20 minutes and removed through the catheter. Recovery rate of ethanol was 82.4%. The maximum blood level of ethanol was obtained 30 to 60 minutes after injection, while the maximum urinary excretion rate was observed after 60 to 120 minutes. The maximum blood levels of ethanol ranged from 0.015 to 0.339 mg/ml. There was a positive correlation between injected volume of ethanol and the maximum blood level (r = 0.66) or the total amount of urinary excretion during 12 hours (r = 0.72). Residual ethanol concentrations in the body were calculated from injected volume and recovery rate of ethanol. Only 2.3% of the residual ethanol in the body was excreted in the urine during the first 12 hours after injection. However, urine/blood ratio of ethanol 1 hour after injection was tremendously high with a wide variation between 1.6 and 230.5. Therefore, a large part of the ethanol absorbed from the cyst wall seems to be excreted directly from the kidney, not entering general circulation. From the estimation of the blood and urine levels of ethanol, it is concluded that 95% ethanol can be applied to the renal cyst wall as a sclerosing agent through the percutaneous ultrasound-guided procedure in the case of good recovery of ethanol.

Changes of contents of CO and water in blood exposed to heat - as to a possibility of estimating pre-exposure CO content of thermally coagulated blood on the basis of its water content.

A blood sample containing CO in a glass vessel was heated in a thermostated water bath at various temperatures for varying lengths of time to compare changes in CO and water contents. With higher temperatures and longer exposure periods the degrees of thermocoagulation advanced and the contents of CO and water decreased. At the individual temperatures examined, changes of CO and water contents showed significant correlations. However, with different temperatures regressions differed significantly. The above was considered to show that the temperature, to which a blood sample had been exposed, must be known for successful application of the water content method to estimate pre-exposure CO content of heat-exposed blood. However, this requirement is considered difficult to be met.

A study on the combined action of CO and HCN in terms of concentration-time products.

Acute toxicity at single and combined exposures of CO and HCN was studied on rats in terms of concentration-time product (ppm . min) necessary to kill animals (lethal CT). The animal was exposed individually to test gas in an animal chamber made of transparent plastics, and test gas was made in gas chamber connected to the animal chamber by a wide and short piece of plastic tube. HCN was produced by addition of NaCN solution to H2SO4 and in case of CO exposure, various amounts of pure CO were introduced. During exposure, gas samples were frequently taken. After exposure, blood sample was withdrawn from the right side of the heart. CO concentrations in the gas and blood were determined gas chromatographically. HCN in the gas sample was measured spectrophotometrically, after being absorbed into NaOH solution in a glass vessel devised by our laboratory. At single exposures, mean lethal CT for CO was 78,000 +/- 22,000 and for HCN was 4,700 +/- 940. In combined exposure, various combinations of CO and HCN were used. A fractional CT, defined as a ratio of CT to lethal CT, multiplied by 100, was calculated for each gas. A linear relationship between fractional CTs of HCN and CO was considered to show a simple additive action between the two gases. The sum of both fractional CTs averaged 100 +/- 26. On the other hand, linear relation was not observed between blood levels of the two toxicants at death.

A blood cyanide distribution study in the rabbits intoxicated by oral route and by inhalation.

Blood cyanide concentration was determined in rabbits intoxicated orally or by inhalation. Experiments were carried out under urethane anaesthesia. In the inhalation experiments, rabbits inhaled a combustion product containing HCN via the tracheal cannula and in the oral studies animals were administered NaCN solution into the stomach. In addition to the carotid artery and jugular vein blood samples, postmortem samples were obtained from both sides of the heart and the descending vena cava. The arterial cyanide concentration in the inhalation group showed a close relationship with ventilation. After an initial rise, blood levels decreased a little, in some cases with transient apnea. At the last stage it again increased with gasping, reaching its maximal value. After ultimate apnea, the blood cyanide concentration declined. The blood cyanide values were higher in the oral group than in the inhalation group. The difference between the two groups became larger in the inferior order, the left heart blood--the right heart blood--blood in the descending vena cava. The left heart/right heart ratio of the inhalation group was significantly higher than that of the oral group (1.28+/- 0.28 vs. 0.95+/- 0.09). The coefficient of variation (c.v.) of the inhalation group was larger than that of the other group. Within the inhalation group, the left heart blood showed the largest c.v. values and this was probably due to redistribution of the cyanide by bloodstream after attainment of the maximal concentration.