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Biomedical subjects

J Nichols

Publications and source records attributed to J Nichols.

At least 19 recordsLinked to original sources

Transcription factors, translocations, and leukemia.

The frequent occurrence of TF gene involvement in translocations associated with leukemia is remarkable, although not yet explained. The wide variety of TFs involved in these translocations and the different stages of cellular maturation argue against a unifying mechanism. Recombinases, active during B-cell and T-cell development, have been implicated in gene arrangements involving TCR genes and in the SIL/SCL rearrangement, which involves two genes not normally rearranged. However, other mechanisms must clearly be active in generating these molecular abnormalities and perhaps they relate to the multistep maturation and differentiation processes and continuous cell turnover seen in hematopoietic cells. The difficulties in obtaining human solid tumor samples may make it more difficult to identify translocations involving TF genes in solid tumors. Recently, the cytogenetic analysis of solid tumors has improved and specific cytogenetic abnormalities have been associated with specific types of tumors. With advanced techniques, such as fluorescent in situ hybridization (a technique that does not depend on cell growth) and PCR, abnormalities involving TF genes will be discovered. Abnormalities of TF genes, other than translocations, have been seen in a broad variety of nonhematopoietic malignancies. The p53 protein has been shown to bind DNA in a sequence-specific fashion and interact with a variety of DNA tumor virus oncoproteins. The broad range of cell types that harbor p53 abnormalities suggests that TF abnormalities will likely be implicated in many solid tumors. We have detailed several examples of how gene rearrangements that accompany chromosomal translocations in acute leukemia can alter the expression or activity of cellular TFs. Several translocations generate fusion RNA transcripts and fusion TF proteins with altered functional characteristics. Other translocations result in the expression of a gene not normally detectable in hematopoietic cells or alter the level of its expression, or affect the promoter usage or exon structure of the gene (Table 2). Studies are underway in many laboratories to characterize the biologic activity of these abnormal TFs and it remains to be proven that these molecular abnormalities are directly linked with leukemogenesis. The identification of abnormal fusion transcripts and proteins may allow specific therapies to be directed against "tumor-specific" DNA, mRNA, or protein targets. Therapeutic strategies based on antisense or ribozyme technology may be used to turn off expression of these genes and inhibit leukemia cell growth. Immunologic methods can also be used to direct therapy against the malignant cells.

Chromosomes, Human

Interleukin 2 receptor targeted fusion toxin (DAB486-IL-2) treatment blocks diabetogenic autoimmunity in non-obese diabetic mice.

Insulin-dependent diabetes mellitus (IDDM) is strikingly similar in the non-obese diabetic (NOD) mouse and humans. In IDDM, the systematic autoimmune destruction of insulin-producing beta cells within the pancreas is dependent on autoreactive T cells. This autoimmune process can be accelerated by transferring spleen cells from diabetic donors into irradiated syngeneic NOD mice. In a previous study we established that interleukin 2 receptor (IL 2R)-bearing cells propagated from pre-diabetic NOD mice promote IDDM. Therefore, we reasoned that specific elimination of IL 2R+ T cells should abort the diabetogenic process. T cell expressing IL 2R can be selectively destroyed with a diphtheria toxin-related IL 2 fusion protein (DAB486-IL-2). We set DAB486-IL-2 the challenging task of preventing fulminant IDDM accelerated by the adoptive transfer of diabetic spleen cells. Eight weeks after the adoptive transfer only 10% and 20% of NOD mice treated with 10 and 5 micrograms/day of DAB486-IL-2, respectively, became diabetic while 100% control mice (vehicle buffer) became diabetic within 5 weeks. A dose of 1 microgram/day of DAB486-IL-2 had no protective effect. Although the protection conferred by DAB486-IL-2 is not permanent, it is maintained for at least 4 weeks following cessation of treatment. Furthermore, even though these NOD mice do eventually become diabetic, the tempo of expression and severity of diabetes, as assessed by the level of hyperglycemia, is dramatically reduced. Although histologic examination of pancreas revealed minimal degree of mononuclear infiltrate within the islets in both groups, the vehicle control mice had fewer islets per section indicating many islets had already been destroyed. In addition, spleen cells from diabetic NOD mice which were pre-treated with DAB486-IL-2 (10 micrograms/day) for 1 week lost their ability to transfer disease. Taken together, these studies strongly support the concept that IL 2R-bearing T cells are essential for the induction of IDDM and suggest that DAB486-IL-2 would be a promising therapeutic approach in the treatment of human IDDM.

Animals

Differentiation inhibiting activity (DIA/LIF) and mouse development.

Analysis of the differentiation in culture of murine embryonic stem (ES) cells has resulted in the identification and characterization of the regulatory factor differentiation inhibiting activity (DIA). DIA specifically suppresses differentiation of the pluripotential ES cells without compromise of their developmental potential. DIA is identical to the pleiotropic cytokine leukaemia inhibitory factor (LIF) which has a broad range of biological activities in vitro and in vivo. It is produced in both diffusible and matrix-localised forms whose expression is differentially regulated. The compartmentalization of DIA/LIF and the modulation of its expression during stem cell differentiation and by other cytokines may be significant elements in the control of early embryo development. These features may also indicate general principles of the regulatory networks which govern stem cell renewal and differentiation in later development.

Animals

Salvage radiotherapy in recurrent Hodgkin's disease.

Forty-four patients with Hodgkin's disease (HD) which relapsed after chemotherapy were treated with salvage radiotherapy (S-RT) with curative intent. Patients were aged 7 to 80 years (median 32 years) at the time of S-RT and the median follow-up from S-RT was 5 years (1-15). Nine patients had recurrent HD following first-line chemotherapy and thirty five patients had refractory HD. Salvage therapy consisted of radiotherapy alone in 25 and combined chemotherapy and radiotherapy in 19 patients. The overall CR rate of salvage therapy was 66%. The overall median survival of 44 patients was 4.6 years from S-RT with 46% 5 year and 40% 10 year survivals. Age (greater than 40 years) and progression free interval (less than or equal to 1 year) were adverse independent prognostic factors for survival on multivariate analysis. The 5 and 10 year progression free survivals were 38% and 23% respectively. Adverse independent prognostic factors for progression-free survival were extranodal site of recurrence and short progression free interval (less than or equal to 1 year). We conclude that radiotherapy with or without chemotherapy has a role in the salvage of patients failing chemotherapy, particularly in those with nodal disease and progression-free interval greater than 1 year.

Adolescent

Inhibition of HIV-1 RNA production by the diphtheria toxin-related IL-2 fusion proteins DAB486IL-2 and DAB389IL-2.

Productive infection of cells by human immunodeficiency virus type 1 (HIV-1) is associated with the activation state of the cell and its obligatory expression of the interleukin-2 receptor (IL-2R), the latter providing a new target for antiviral therapy. A quantitative RNA-RNA hybridization assay is employed to detect production of HIV-1 RNA and to show that two IL-2 diphtheria toxin-related fusion proteins (DAB486IL-2 and its more potent, truncated form DAB389IL-2) inhibit HIV-1 RNA production in infected cells. A mutant form of DAB486IL-2 containing a single point mutation that inactivates the adenosine diphosphate ribosyltransferase activity of the toxin does not inhibit HIV RNA production, even though the molecule binds to the IL-2R. The active fusion proteins inhibit viral RNA replication in cells infected with HIV-1 clinical isolates as well as with a ZDV-resistant strain of HIV-1. These results indicate that IL-2 receptor-targeted fusion proteins can be utilized to inhibit HIV-1 replication effectively in infected human lymphocytes.

Antiviral Agents

An investigation of the fate of cells transplanted orthotopically between morulae/nascent blastocysts in the mouse.

A technique has been devised for selectively replacing some inside cells (ICs) or outside cells (OCs) of late morulae/nascent blastocysts with corresponding cells from genetically dissimilar, synchronous embryos. The main purpose was to determine whether the inner cell mass (ICM) contributes cells to the overlying polar trophectoderm at any stage during the blastocyst phase of development. Notwithstanding the high incidence and level of chimaerism in ICM derivatives of post-implantation conceptuses obtained in the IC transplantation experiments, trophoblast tissue was composed entirely of host cells in the majority of cases. Even where a donor IC contribution to trophoblast was detected there were strong grounds for suspecting that it was due to tissue contamination rather than genuine chimaerism. Thus, not only did such contributions differ in both level and distribution from those produced by transplantation of OCs but they also varied markedly in frequency according to the day of gestation on which conceptuses were dissected. The possibility that ICs regularly colonize the polar trophectoderm but fail to persist there was excluded by the results of short-term transplantation experiments using an in-situ genetic marker. These findings offer no support for the hypothesis that the ICM serves as a population of stem cells for the trophectoderm as well as the primitive endoderm and ectoderm during normal development. The frequency of chimaerism was lower in OC than IC transplantation experiments. Nevertheless, in a substantial proportion of chimaeras, OCs colonized derivatives of the ICM. This is consistent with evidence from other studies that some outside cells divide differentially at 5th cleavage to produce an OC plus an IC rather than two OCs.

Animals

Murine spontaneous T-cell leukemia constitutively expressing IL-2 receptor--a model for human T-cell malignancies expressing IL-2 receptor.

We describe a new, spontaneously occurring BALB/c-derived murine T-cell leukemia. The leukemic cells, designated LB, grow rapidly and progressively in the syngeneic host with no signs of effective immunological resistance. LB cells expressed the Thy-1+, Lyt-2+, L3T4-, CD3- class-I+, CD25+ (IL-2 receptor, IL-2R), class-II-, gp70- phenotype. As LB cells express IL-2, as indicated by staining with 2 distinct anti-CD25 IL-2R monoclonal antibodies (MAbs), the therapeutic efficacy of IL-2-diphtheria toxin-related protein was tested on this leukemic model. IL-2-diphtheria toxin, but not diphtheria toxin, efficiently inhibited the proliferation of LB cells. The proliferation of a murine myeloma cell line, which does not express IL-2R, was not inhibited by IL-2-diphtheria toxin. The possible implantation of this animal model in fundamental and practical studies is discussed.

Animals

Pharmacokinetics of the recombinant fusion protein DAB486IL-2 in animal models.

The kinetics of the in vitro cytotoxicity of DAB486IL-2, a genetically engineered fusion protein containing a portion of diphtheria toxin and human interleukin-2, were examined in the C91/PL cell line, which constitutively expresses IL-2 receptors. Maximal inhibition of protein synthesis was observed by 4-6 h after DAB486IL-2 addition at a concentration of 300 ng/ml. The tissue distribution, urinary excretion, and plasma pharmacokinetics of DAB486IL-2 in the rat and its plasma pharmacokinetics in the monkey were also examined. In rats the primary site of distribution of [35S]-DAB486IL-2 outside the vasculature appears to be the liver, followed by the kidney, spleen, and lung. Persistence of radioactive material in the liver and urinary excretion of metabolic degradation products suggest that labeled protein is metabolized by hepatic tissue. Following i.v. bolus administration of DAB486IL-2, the initial serum half-life for both the rat and the monkey was approximately 5 min. The overall clearance rate of drug for the two species differed, with DAB486IL-2 being cleared from circulation 2-3 times more rapidly in the monkey. Presence of high levels of neutralizing antibodies to diphtheria toxin in the rat significantly influenced the clearance of bioactive DAB486IL-2. However, the question as to whether the presence of in vitro biological activity for the molecule is masked by the presence of antibodies cannot be clearly answered.

Animals

Developmentally programmed induction of differentiation inhibiting activity and the control of stem cell populations.

Differentiation inhibiting activity/leukemia inhibitory factor (DIA/LIF) is a glycoprotein that controls differentiation of pluripotential stem cells. Alternative transcription generates both diffusible and matrix-associated forms of DIA/LIF. Transcriptional analysis using a sensitive ribonuclease protection assay revealed that the two messages are expressed independently, consistent with the proposition that the two forms of DIA/LIF have distinct biological roles. DIA/LIF expression was found to be activated early during differentiation of embryonic stem (ES) cells, providing a mechanism for feedback regulation of stem cell renewal. Expression of DIA/LIF by mesenchymal cells was shown to be controlled in a paracrine manner by polypeptide regulatory factors. Specific expression of the two forms of DIA/LIF was also demonstrated in the egg cylinder-stage mouse embryo. The combination of cell type-specific and signal-specific regulation enables very precise control over DIA/LIF expression and may represent an important component of the regulatory networks that govern stem cell proliferation and differentiation during mammalian development.

Animals

Establishment of germ-line-competent embryonic stem (ES) cells using differentiation inhibiting activity.

The regulatory factor Differentiation Inhibiting Activity/Leukaemia Inhibitory Factor (DIA/LIF) suppresses the differentiation of cultured embryonic stem (ES) cells. In the present study, it is shown that ES cell lines can be derived and maintained in the absence of feeder layers using medium supplemented with purified DIA/LIF. These cells can differentiate normally in vitro and in vivo and they retain the capacity for germ-line transmission. DIA/LIF therefore fulfils the essential function of feeders in the isolation of pluripotential stem cells.

Animals

Effect of damage to the zona pellucida on development of preimplantation embryos in the mouse.

The effect on development of early mouse embryos of making a hair-line slit in the zona pellucida of approximately one-third its diameter was investigated. The rate of development to mid-gestation of operated zygotes and 2-cell embryos transferred directly to the oviduct was significantly lower than that of sham-operated or unoperated controls. However, the operation had no discernible effect on the development of 2-cell embryos that were cultured for 2 days prior to transfer to the uterus, or on embryos composed of 8 or more cells transferred directly to the oviduct. Zona slit zygotes and 2-cell embryos exhibited a significantly higher rate of anomalous development to the morula or blastocyst stage than controls following short-term transfer to the adult or immature oviduct. Such anomalies could not be attributed to damage of the embryos by leucocytes or bacteria entering through the wound in the zona. Rather, the typically non-spherical shape of slit zonae, together with the fact that some were empty on recovery, was consistent with operated embryos having been damaged by compression during passage through the oviduct. This suggests that, providing it is intact, the zona pellucida protects the early embryo from contraction of the oviductal musculature which is sufficient to lyse, arrest or extrude blastomeres prior to the formation of intercellular junctions. Hence, in experimental manipulations entailing damage to the zonae of early embryos, there may be a case for allowing them to form morulae in vitro prior to transfer, rather than returning them directly to the oviduct.

Animals

The effect of low impact dance training on aerobic capacity, submaximal heart rates and body composition of college-aged females.

The purpose of this study was to examine the effects of a 12 week program of low impact aerobic dance conditioning on VO2max, submaximal heart rates and body composition of college-aged women. Sixteen women exercised three times per week for approximately 45 minutes per session at 75-85% of their heart rate reserve. VO2max was measured by indirect calorimetry using a treadmill protocol. Submaximal heart rates were measured by electrocardiography, and body fat was assessed by hydrostatic weight. All testing was conducted within one week pre- and posttraining. Training sessions consisted of a 5-10 minute warm up, 30-35 minute low impact aerobic dance segment and a 5 minute cool down. Posttest results revealed a small (7%), but significant increase in VO2max (pre: 38.3 ml/kg/min; post: 41.3 ml/kg/min, X +/- SD, p less than 0.05). Submaximal heart rates at minutes 2-3, 3-4 and 4-5 of the graded exercise test decreased significantly. Body fat decreased from 25 +/- 6.8% to 21 +/- 6.3% (p less than 0.01) with no posttraining change in body weight. It was concluded that low impact aerobic dance is as effective as other endurance training regimens in improving cardiovascular fitness and decreasing body fat.

Adipose Tissue

Thermic effect of food at rest and following swim exercise in trained college men and women.

The calorigenic effect of a liquid meal was studied in 8 (5 male, 3 female) lean, trained swimmers to assess the effect of swim exercise on dietary-induced thermogenesis. Metabolic rate was measured by indirect calorimetry under 3 conditions: (A) 4 h postexercise in the postabsorptive state; (B) 4 h of rest in the postprandial state, and (C) 4 h postexercise in the postprandial state. At 20 min postexercise metabolic rate had increased 22% over RMR; however, by 40 min postexercise it had returned to the preexercise level. Following both conditions B and C oxygen consumption (VO2) remained significantly elevated for 3 h. The mean rise in caloric expenditure was 24.8 kcal/h when exercise preceded the meal, compared to 20.2 kcal/h when exercise was not performed. This difference of approximately 18 kcal/4 h, although of negligible importance in short-term energy balance, could be significant in long-term energy regulation, and thus contribute to body weight control.

Adipose Tissue

Food intake at age 8. 1. Energy, macro- and micronutrients.

The food energy, macro-, and micronutrient intake of 78 boys and 63 girls aged 8 years who have been studied since birth is reported. Nutrient intake was calculated from 4 day weighed records using the CSIRONET data bank based on British tables of food composition. The mean values for daily intake for girls and boys, respectively, for energy (MJ), protein, fat, carbohydrate and fibre (g) were 7.4 and 6.5; 64 and 57; 71 and 62; 232 and 206; and 15 and 12 g. Half the food energy came from carbohydrate, divided equally between starch and simple sugars, 15% from protein, and 35% from fat. This comprised 41% saturated (S), 33% mono- and 16% polyunsaturated (P). The mean P:S ratio for girls was 0.46, and 0.41 for boys. Overall sweets and soft drinks contributed 4-5% to energy intake, with large individual differences. Mineral and vitamin intakes were satisfactory in relation to theoretical optimal population needs. Vitamin and/or mineral supplements were taken by 8.5% of the sample. These results are discussed in relation to the Nutritional Guidelines.

Australia

Food intake at age 8. 2. Frequency, company and place of meals.

In a sample of 141 healthy 8 year old children, over 95% had three main meals on each of the 4 days of the food record. Two-thirds had an afternoon snack and nearly one-half a morning snack on each of the 4 days, but only one-quarter had supper on each day. Seventy-one per cent of all meals were eaten at home with the family, 20% at school with friends, and less than 1% of meals were eaten at a restaurant or fast food outlet. These data show that amongst children of primary school age, take away sources of food make a very small contribution to the diet, and that eating habits at home are of greatest importance to the child's diet.

Child