Inositol lipids and phosphates in the differentiation of blood cells.
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Biomedical subjects
Publications and source records attributed to G Brown.
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Direct dose-dependent effects of angiotensin II on renal tubular sodium reabsorption have been demonstrated. Alterations in tubular sodium reabsorption may occur via modulation of renal Na,K-ATPase activity. Thus, these experiments were undertaken to ascertain whether angiotensin II could influence renal cortical Na,K-ATPase activity. Angiotensin II, 495 ng/microliters/h, or vehicle (controls) was infused for 24 h via miniosmotic pumps 48 h after rats were adrenalectomized and implanted with osmotic pumps containing 12.5 micrograms/microliters corticosterone (Treatment I) or both corticosterone and 0.2 microgram/microliter aldosterone (Treatment II), and in rats receiving 3% NaCl in their food (sodium loaded, Treatment III). Rats receiving Treatments I and III received saline to drink. Renal cortical microsomal membranes were prepared, and the effects of angiotensin II infusion on the K1/2 and Vmax for Na, K, and ATP determined. Angiotensin II infusions were associated with (i) a decrease (P less than 0.001) in the K1/2 for Na activation of Na,K-ATPase from 14 +/- 3 to 6 +/- 1 (n = 4 experiments), 16 +/- 1 to 12 +/- 1 (n = 5), and 12 +/- 3 to 7 +/- 1 (n = 5) mM (means +/- SE) for treatments I, II, and III, respectively; (ii) no changes in the K1/2 for K activation or the Km for ATP; (iii) no changes in the Vmax for Na, K, or ATP; and (iv) no change in Mg-ATPase activity. We conclude that angiotensin II infusion is associated with a decrease in the K1/2 of renal cortical Na,K-ATPase activity for sodium. This action of angiotensin II on the enzyme activity may contribute to the regulation of tubular sodium transport.
We analyzed the clinical courses of 94 patients with treated primary hypertension and initially normal serum creatinine concentrations (less than or equal to 133 mumol per liter [less than or equal to 1.5 mg per deciliter]) who were followed for a mean (+/- SD) of 58 +/- 34 months (range, 12 to 174) to determine the frequency with which renal function deteriorated and the factors associated with deterioration. Fourteen patients (15 percent) had an increase in serum creatinine concentrations (greater than or equal to 35 mumol per liter [greater than or equal to 0.4 mg per deciliter]); in 16 percent of the 61 patients with apparently good control of blood pressure, the serum creatinine concentration rose 59 +/- 33 mumol per liter (0.67 +/- 0.38 mg per deciliter). Despite good control of diastolic blood pressure (less than or equal to 90 mm Hg), black patients were twice as likely as white patients to have elevations in serum creatinine (23 percent vs. 11 percent). Stepwise discriminant function analysis showed that a significant rise in the serum creatinine concentration was most likely to occur in association with older age, black race, a higher number of missed office visits, and employment as a laborer. We conclude that although renal function was preserved in 85 percent of patients with treated hypertension, it may deteriorate in some patients despite good blood-pressure control. Our observations may partly explain why hypertension, particularly among black persons, remains a leading cause of renal disease in the United States.
A 9-point clinical rating scale was used to assess the severity of hopelessness in 141 patients hospitalized with suicidal ideation. The patients were followed from 5 to 10 years, and 10 (7.1%) eventually committed suicide. The mean hopelessness rating for the patients committing suicide was significantly higher than that for the patients not committing suicide. A cutoff score of 6 or above successfully predicted 9 (90.0%) of those committing suicide. The results supported previous findings in which self-reported hopelessness on the Beck Hopelessness Scale was reported to predict suicide in both psychiatric outpatients and inpatients.
We investigated the degree of content specificity evident in the negative cognitions associated with anxiety and depression in two large samples of general psychiatric outpatients. Standardized measures of affect and cognition were analyzed in a multiple regression design. As predicted by Beck's (1967, 1976, 1987) cognitive theory of psychopathology, thoughts of loss and failure were specifically associated with depression, whereas cognitions of harm and danger were uniquely predictive of anxiety. In addition, hopelessness was specific to depression and not to anxiety. Dysfunctional beliefs showed no consistent association with either mood state. The implication of these results, as well as related findings, is discussed in terms of a cognitive perspective on the differentiation of emotional disorders. Also discussed are the methodological difficulties encountered in research on cognitive-affective relationships.
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Steele and others have suggested that minimizing changes in glucose specific activity when estimating hepatic glucose output (HGO) during glucose infusions could reduce non-steady-state errors. This approach was assessed in nondiabetic and type II diabetic subjects during constant low dose [27 mumol.kg ideal body wt (IBW)-1.min-1] glucose infusion followed by a 12 mmol/l hyperglycemic clamp. Eight subjects had paired tests with and without labeled infusions. Labeled infusion was used to compare HGO in 11 nondiabetic and 15 diabetic subjects. Whereas unlabeled infusions produced negative values for endogenous glucose output, labeled infusions largely eliminated this error and reduced the dependence of the Steele model on the pool fraction in the paired tests. By use of labeled infusions, 11 nondiabetic subjects suppressed HGO from 10.2 +/- 0.6 (SE) fasting to 0.8 +/- 0.9 mumol.kg IBW-1.min-1 after 90 min of glucose infusion and to -1.9 +/- 0.5 mumol.kg IBW-1.min-1 after 90 min of a 12 mmol/l glucose clamp, but 15 diabetic subjects suppressed only partially from 13.0 +/- 0.9 fasting to 5.7 +/- 1.2 at the end of the glucose infusion and 5.6 +/- 1.0 mumol.kg IBW-1.min-1 in the clamp (P = 0.02, 0.002, and less than 0.001, respectively).
The revised Beck Depression Inventory (BDI) was administered to 174 male and 276 female psychiatric outpatients diagnosed with affective disorders. The mean BDI scores, mean number of symptoms claimed, and corrected item-total correlations were comparable for both sexes, and the coefficient alpha for each sex was .88. Principal components analyses found four dimensions of depression underlying both sexes' BDI item-intercorrelation matrices. Although men and women had comparable dimensions with respect to weight loss, self-blame, and somatic-performance symptoms, men had affective and performance symptoms loading on the same factor, whereas women had affective and cognitive symptoms loading on the same dimension.
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A health visitor and a social worker met along with a family centre worker to plan 'a working group for parents within the community who were perceived to have difficulty in appreciating and providing for the needs of their children and who had themselves had an impoverished nurturing experience'. The idea was to establish a group of lone parents who were making frequent clinic contacts to run for 6-8 weeks. It was to look at issues such as trust, self-esteem, parenting experience, expectations and sexuality. The group achieved its modest aims resulting in both a dropping off of clinic attendances and a less anxious social services profile in most of the women's lives.
Various data in the literature suggest that progenitor cells of monocytes and B cells are closely related. In this study we have investigated this notion by using two-dimensional gel electrophoresis patterns of total cellular phosphoproteins to assess the similarity and thus the close relationships between erythroid, granulocyte, monocyte, and B-lymphocyte cell lines that typify immature cells of these lineages. In previous studies, six proteins had been identified whose constitutive phosphorylation correlated with the capacity of HL60 variant cell lines to differentiate towards either neutrophils (four proteins) or monocytes (two proteins). The presence or absence of five of these phosphoproteins in autoradiographs obtained for the pre-B-cell lines Nalm6 and SMSB showed that the pre-B-cell lines most closely resembled lines able to differentiate towards monocytes (HL6015-12, HL60M2, U937, and ML-1) as opposed to lines restricted to neutrophil (HL60Ast3) or erythroid differentiation (K562). The K562 phosphoprotein pattern resembled that obtained for HL60Ast3. Progressive changes in the constitutive phosphorylation of the five proteins were observed suggesting that cells that have acquired the potential for either neutrophil, monocyte, or B-cell differentiation have initially diversified in a linear progressive manner. This observation supports a model for hemopoiesis that suggests that, during progenitor cell development, differentiation potentials are expressed individually in the above order. Two additional phosphoprotein spots were found to be restricted to the pre-B-cell lines. These phosphoproteins, together with those that change their intensity in autoradiographs of erythroid, myeloid, and B-cell lines, suggest that protein phosphorylation plays an important role during cell diversification.
A new method for isolating cell nuclei is described which involves freezing and thawing cells in 2% Tween 40, then gentle homogenization to release nuclei, followed by immediate microcentrifugation through 50% sucrose. Purified nuclei were obtained in 3 min and yields of 78-95% were obtained from a variety of human hemopoietic cells. Electron microscope analysis of nuclei obtained from HL60 cells showed that 89% of the nuclei were intact and have an appropriate morphology. A low level of contamination with other organelles was revealed by electron microscopy and by using specific assays for plasma membrane, mitochondria, lysosomes, Golgi membrane, and endoplasmic reticulum (0.5-5.5%). The value of the technique is that nuclear proteins and small metabolites which might be lost by rapid leakage from isolated nuclei and the possibility of biochemical modification of cellular constituents are minimized by using a rapid isolation procedure.
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We have investigated the possible relationship between c-myc gene activity and other variable traits in HL60. In a panel of variant lines, a good correlation was observed between myc gene copy number and the level of myc mRNA. There was no correlation between myc amplification or expression and the resistance of the lines to induction of terminal neutrophilic or monocytic differentiation. Therefore, myc mRNA level does not appear to determine the ability of the variant HL60 lines to respond to inducers of differentiation. Flow cytometric analyses of the expression of a differentiation antigen (AGF 4.36) revealed stable negative and positive subpopulations in growing HL60. myc amplification, expression, and inducibility were identical in these subpopulations, suggesting that variation of these traits in HL60 sublines and variants is not due to maturation state differences. myc gene copy number was also identical in transferrin receptor positive (proliferating) and negative (resting) populations. These data contradict the notion that myc amplification has been important in determining the in vitro biological properties of HL60.
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The human promyeloid cell line H60 can be induced to differentiate towards either neutrophils or monocytes. Variant cell lines, derived from HL60, which show reduced capacities for neutrophil and monocyte differentiation can be arranged in a developmental sequence which suggests that the potentials for neutrophil and monocyte differentiation are expressed sequentially by HL60 cells in this order. Analysis of the patterns of total cellular phosphoproteins within HL60 and 5 variant cell lines, by two-dimensional gel electrophoresis, has identified 6 distinct phosphoproteins which show progressive differences in the intensity of spots between the variant lines. The changes in these phosphoproteins relate to the position of the lines within the proposed development sequence. Similarly, lines placed close together in the sequence are more similar, as regards phosphoprotein profiles, than lines placed far apart. These studies provide direct evidence in favour of the hypothesis that the potentials for neutrophil and monocyte differentiation are expressed sequentially during myelopoiesis. Furthermore, two phosphoprotein spots were found to be restricted to lines able to differentiate towards monocytes. These proteins may play important roles during commitment to monocyte differentiation.
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