Autologous blood stem cell transplantation in acute leukaemia: present status and future directions.
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Biomedical subjects
Publications and source records attributed to A Rice.
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We have studied peripheral blood stem cells (PBSC) collected by cytapheresis following intensive chemotherapy, from 13 patients with acute leukemia, in long term culture (LTC). Peripheral blood was cultured with (n = 10) and without (n = 21) the addition of a preformed, irradiated stromal layer. In this latter LTC our results confirm that peripheral blood is capable of producing CFU-GM and nucleated cells in the absence of the formation of an adherent stromal layer. However, peripheral blood cultured in the presence of an irradiated stromal layer is capable of a significantly higher proliferative response (total production of CFU GM per flask - mean = 57529) than in the absence of an irradiated stromal layer (total production of CFU GM per flask - mean = 26739, p less than 0.03). Our results suggest that PBSC contain a primitive nonplastic adherent cell that requires the presence of a stromal layer for its expression. These findings provide further support for the use of peripheral blood stem cells for autologous transplantation.
Autologous blood stem cell transplantation (ABSCT) is a new technique which has been increasingly used in recent years. It is now well established that ABSCT can be performed as safely as autologous bone marrow transplantation (ABMT) when high numbers of hemopoietic precursors are infused. Multiple leukophoreses are performed during marrow regeneration following chemotherapy-induced aplasia. Hemopoietic recovery (predominantly the granulocytic series) is probably faster after ABSCT than after ABMT. Among other advantages there may be minimal contamination by residual tumor cells in buffy-coats; however, this has not been fully investigated.
Elevated maternal levels of alphafetoprotein (AFP) in midtrimester are believed by some to be a useful screening test for premature labour, low birthweight and low birthweight for gestation. In a prospective study on 887 randomly selected pregnant women we found that although there was an association between low birthweight and elevated AFP, the test would miss five out of every six cases of low birthweight and there would be nine false-positives for every case correctly identified. The test is not therefore sufficiently predictive of low birthweight to be of value as a screening test for this condition.
In the course of an investigation on the effects of thyroid releasing hormone in pregnant women we noted a dramatic increase in the levels of placental protein 5 (PP5) in maternal blood. This increase ranged from 10 times to over 40 times the basal levels. Further study showed that the rise was associated with the use of heparin to maintain the patency of the cannula through which the samples were collected. Furthermore, the phenomenon appeared to be systemic rather than local, and may well be due to a direct effect of heparin on PP5 secretion by the placenta.
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Tritrichomonas foetus and Trichomonas vaginalis are both incapable of de novo purine nucleotide synthesis. Previous studies indicated that T. foetus relies mainly on the salvage of hypoxanthine and subsequent conversion of IMP to AMP and GMP, whereas T. vaginalis depends on direct conversions of exogenous adenosine to AMP and guanosine to GMP without much interconversion between the two nucleotides. These two different types of purine salvage suggest the possibility of differential sensitivities between the two species of trichomonad flagellates toward different purine antimetabolites. Mycophenolic acid, hadacidin, 8-azaguanine, and formycin B inhibited the growth of T. foetus but had no effect on T. vaginalis. Mycophenolic acid acted by blocking conversion of IMP to GMP, hadacidin inhibited conversion of IMP to AMP, and 8-azaguanine was incorporated into the T. foetus nucleotide pool, likely via hypoxanthine phosphoribosyl transferase. Formycin B was converted to 5'-monophosphate in T. foetus and inhibited the conversion of IMP to AMP. Its precise mechanism of action on T. foetus remains, however, to be elucidated. Alanosine, whose ribonucleotide derivative is a potent inhibitor of adenylosuccinate synthetase, had no effect on the growth or hypoxanthine incorporation in T. foetus, which may be due to the lack of conversion of alanosine to the ribonucleotide because of the absence of de novo purine nucleotide synthesis in parasites. Four adenosine analogs, adenine arabinoside, tubercidin, sangivamycin, and toyocamycin, were found inhibitory to the growth of T. vaginalis but showed little effect on T. foetus growth. Further investigations suggested that these four compounds acted on T. vaginalis by blocking incorporation of adenosine into the adenine nucleotide pool.
A partly defined medium was successfully designed for the cultivation of Tritrichomonas foetus, an anaerobic protozoan parasite of cattle. The medium consists of hypoxanthine, uracil, and thymidine as the sole precursors of nucleotides in T. foetus. Elimination of any one of the three precursors from the medium led to cessation of T. foetus growth. The information provided by this medium verifies our previous observations that T. foetus is incapable of de novo purine and pyrimidine synthesis, that hypoxanthine can be converted to AMP and GMP, that uracil is incorporated into all pyrimidine ribonucleotides including UDP-glucose--the precursor of glycogen synthesis, and that thymidine is the only precursor of TMP. The omission of folate from the medium, without affecting growth of T. foetus, also supports our previous finding that the parasite does not have functioning dihydrofolate reductase or thymidylate synthetase. The successful plating of T. foetus on agar plates incorporating the partly defined medium with near 100% plating efficiency makes it possible to isolate T. foetus mutants for further studies of purine and pyrimidine metabolism in this parasite.
We describe a method for the preparation of purified PP5 suitable for use as a tracer ligand in RIA. The method is based on the observation that PP5 will bind to heparin and employs a heparin-Sepharose column as a key step in the procedure.
The anaerobic protozoon Tritrichomonas foetus was found incapable of de novo purine synthesis by its failure to incorporate radiolabeled glycine or formate into the nucleotide pool. It had, on the other hand, high activities in incorporating adenine, hypoxanthine or inosine. Radiolabel pulse-chase experiments indicated that adenine, hypoxanthine and inosine all entered the pool through conversion to IMP. The parasite contained hypoxanthine phosphoribosyl transferase, adenine deaminase and inosine phosphorylase, but no adenine phosphoribosyl transferase, inosine kinase or inosine phosphotransferase activity. Adenine and inosine had to be converted to hypoxanthine before incorporation. Adenosine was also rapidly converted to hypoxanthine in T. foetus cell-free extracts, but the presence of adenosine kinase in the parasite allowed some conversion of adenosine directly to AMP. Guanine and xanthine were directly incorporated into GMP and XMP, probably due to the guanine and xanthine phosphoribosyl transferase. There were also strong enzyme activities which convert guanosine to guanine and guanine to xanthine. A guanosine phosphotransferase was found in the 10(5) X g sedimentable fraction of T. foetus, and was capable of converting some guanosine to GMP. This network of T. foetus purine salvage suggests the importance of hypoxanthine-guanine-xanthine phosphoribosyl transferase activities in the parasite.
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To assess morphine-induced location preferences and flavor aversions, rats were administered morphine sulfate (10 mg/kg, IP) either immediately before (Experiment 1) or immediately after (Experiment 2) confinement for 20 min in one side of a shuttlebox with access to a flavored solution. On control trails the rats were administered saline and confined for 20 min on the opposite side with a differently flavored solution. In subsequent choice tests, it was found that morphine injections before confinement produced a preference for the side associated with morphine and indifference to the flavors, whereas morphine injections after confinement produced an aversion to the flavor paired with morphine and indifference to the sides. Experiments 3 and 4, using a procedure similar to that of Experiment 1, showed that naloxone (1 mg/kg, IP) blocked the morphine-induced side preference, although given alone it was without effect in this test.
We outline methods for identifying DNAs containing sequences complementary to specific mRNAs, and provide a number of complementary approaches for mapping the arrangement of mRNAs along the DNA. These methods, together with S1 nuclease mapping and the direct visualization of R-loops with the electron microscope, provide a comprehensive approach to defining the architecture of mRNAs coding for specific polypeptides and the arrangement of RNA transcripts along the genome. This detailed cartographic information can then be used to study the steps in the processing of mature mRNAs and determine the modes by which the expression of specific genes is regulated.
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