Citations

Abstract In a recent paper Howard (1932) claims to have shown, by three methods, that the "expected" osmotic equilibria exist between the yolk and white of a hen's egg. Johlin (1933) has criticized her technique of cryhydric measurement and re-asserted that the yolk and white of an egg hive different values for depression of freezing point. Although Needham (1931) and Meyerhof (1931) have considered the possibility of the outer layer of yolk having a lower osmotic pressure than the inner, Howard gives no experimental evidence indicating the existence of an osmotic gradient within the yolk. The methods she used being apparently incapable of showing the difference in osmotic pressure between the whole yolk and the whole white of an egg were presumably unable also to detect the osmotic gradient in the yolk. Grollman's (1931) criticisms of the Hill thermo-electric method for the measurement of vapour pressures when employed with viscous solutions were repeated by Howard, with no other evidence than that it gave results which disagreed with her own. In particular, Bateman's low vapour pressure depression found in mixtures of egg yolk and egg white are declared to be incompatible with high vapour pressure depressions for yolk. It is strange that in the differentiation of the properties of egg yolk and white so many authors should have considered the yolk as homogeneous. It is a well-known fact that the formation of an egg yolk occurs by daily deposits in the ovary of the hen. These extend over several days and that the integrity of the daily deposit is maintained more or less for many days is evidenced by observation of the spherical zones in the yolk of a frozen egg that has been sectioned. Also it is easy to withdraw from the centre of the yolk, using a fine pipette, white yolk which is different chemically from the surrounding yellow yolk. Since no membrane is known to separate these two kinds of yolk nor the daily deposit of yolk, the existence of this non-homogeneity within the yolk must be an indication of the slowness of equilibration inside a hen's egg. Hence, when one speaks of the difference in osmotic pressure of average egg white and average egg yolk, no conclusions can be drawn logically regarding the difference in osmotic pressure on opposite sides of the vitelline membrane. As the Hill thermoelectric method of measuring vapour pressure requires but small quantities of solution, it was of interest to use this micro method to study the difference in osmotic pressures of samples of yolk and white obtained on opposite sides of the membrane.

Abstract Several types of adrenocortical tumors that lead to Cushing syndrome may be caused by aberrant cyclic AMP (cAMP) signaling. We recently identified patients with micronodular adrenocortical hyperplasia who were carriers of inactivating mutations in the 2q-located phosphodiesterase 11A (PDE11A) gene. We now studied the frequency of two missense substitutions, R804H and R867G, in conserved regions of the enzyme in several sets of normal controls, including 745 individuals enrolled in a longitudinal cohort study, the New York Cancer Project. In the latter, we also screened for the presence of the previously identified PDE11A nonsense mutations. R804H and R867G were frequent among patients with adrenocortical tumors; although statistical significance was not reached, these variants affected significantly enzymatic function in vitro with variable increases in cAMP and/or cyclic guanosine 3′,5′-monophosphate levels in HeLa and HEK293 cells. Adrenocortical tissues carrying the R804H mutation showed 2q allelic losses and higher cyclic nucleotide levels and cAMP-responsive element binding protein phosphorylation. We conclude that missense mutations of the PDE11A gene that affect enzymatic activity in vitro are present in the general population; protein-truncating PDE11A mutations may also contribute to a predisposition to other tumors, in addition to their association with adrenocortical hyperplasia. We speculate that PDE11A genetic defects may be associated with adrenal pathology in a wider than previously suspected clinical spectrum that includes asymptomatic individuals. (Cancer Res 2006; 66(24): 11571-5)

Abstract Among the best-characterized genetic alterations in gliomas is the amplification of the epidermal growth factor receptor (EGFR) gene, present in ∼40% of glioblastoma multiforme, and frequently associated with the EGFRvIII gene rearrangement. We have previously shown that attenuated vaccine strains of measles virus have potent antitumor activity against gliomas, and identified H protein mutations, which ablate recognition of the natural measles virus receptors CD46 and SLAM. Retargeted recombinant viruses were generated from the measles Edmonston-NSe vaccine strain displaying a single-chain antibody against EGFRvIII at the COOH terminus of H and containing the marker green fluorescent protein (GFP) gene in position 1. Two different H mutants were employed: HSNS (V451S, Y481N, and A527S)-CD46 blind, and HAA (Y481A and R533A)-CD46 and SLAM blind. MV-GFP virus was used as a positive control. Both EGFRvIII-retargeted viruses had significant antitumor activity against EGFRvIII-expressing glioblastoma multiforme but no cytopathic effect against normal cells. In an orthotopic model of EGFRvIII-expressing GBM39 xenografts, there was comparable therapeutic efficacy between retargeted strains and unmodified MV-GFP and statistically significant prolongation of survival in treated animals compared with the control group (P = 0.001). Formation of syncytia was observed in tumors treated with retargeted viruses, with a surrounding infiltrate consisting of macrophages and natural killer cells. In summary, EGFRvIII-retargeted oncolytic measles virus strains have comparable therapeutic efficacy with the unmodified MV-GFP strain against EGFRvIII-expressing glioma lines and xenografts with improved therapeutic index, a finding with potential translational implications in glioma virotherapy. (Cancer Res 2006; 66(24): 11840-50)

Abstract In the smooth muscle of guinea-pig uterus (oestrogen dominated) both noradrenaline (Nor) (α-action) and acetylcholine (ACh) depolarized the membrane, reduced membrane resistance, accelerated spike discharge and increased isometric tension proportionately. The magnitude of the response to ACh was directly related to the dose of ACh, the duration remaining that of the period of drug application. The response to Nor was slow in onset and outlasted the period of drug application by several minutes. Increasing doses of Nor mainly prolonged the duration of the response. The magnitude of the response depended on whether the depolarization by Nor reached firing threshold. The phasic tension response to evoked spikes was increased by Nor. Cooling reduced, and at 6°C abolished the depolarization caused by Nor but did not significantly change the response to ACh. The results obtained with variations of external potassium and chloride indicate that both ACh and Nor increased the membrane permeability to potassium, but that the Nor effect was small and shortlasting. Chloride was not involved in the depolarization caused by ACh, while the magnitude of depolarization and of the increase in membrane conductance by Nor were a direct function of the external chloride concentration. The removal of sodium from the external medium slowed the relaxation of the phasic tension responses to evoked spikes and increased resting tension. In the absence of spikes the tissue relaxed. The depolarization and reduction of membrane resistance by ACh were directly related to the external sodium concentration regardless whether sucrose or tris-Cl were used as substitute. The depolarization and reduction of membrane resistance by Nor did not depend on external sodium (tris substitution) though in the simultaneous absence of sodium and chloride (sucrose substitution) the response to Nor was abolished. The results obtained when NaCl was replaced with tris-Cl suggest that ACh may, in the absence of sodium, increase the entry of calcium. The failure of calcium removal from the cytoplasm may explain the maintained contracture. The contracture produced by Nor was increased in the absence of sodium, suggesting that, in addition to an increased calcium entry from outside, there may have been a release of sequestered calcium.

Abstract The α action of catecholamines on oestrogen dominated guinea-pig uterus is stimulant. The cell membrane is depolarized, membrane conductance is increased, spike discharge is accelerated and tension develops. This action resembles that of acetylcholine though catecholamines are less potent, and, in equiactive concentrations, catecholamines have a longer latency and a longer duration of action. Evidence, obtained by modifications of the ionic environment, indicates that the depolarization by acetylcholine is due to an increase in sodium and calcium permeability and that acetylcholine can release calcium from intracellular stores. The depolarization by catecholamines is due to an increase in chloride permeability and, in addition, sodium is required for the ensuing increase of spike discharge. Catecholamines produce an increase in the force of contraction, long outlasting their immediate stimulation. Moreover, their effect on membrane potential and membrane conductance persists in the presence of lanthanum. These results suggest that Ca release from intracellular stores may be the primary effect produced by the α action of catecholamines and that the increase in the cytoplasmic Ca2+ concentration may cause the changes at the cell membrane.

We present an efficient feature selection algorithm for computer aided detection (CAD) computed tomographic (CT) colonography. The algorithm (1) determines an appropriate piecewise linear network (PLN) model by cross validation, (2) applies the orthonormal least square (OLS) procedure to the PLN model utilizing a Modified Schmidt procedure, and (3) uses a floating search algorithm to select features that minimize the output variance. The undesirable "nesting effect" is prevented by the floating search approach, and the piecewise linear OLS procedure makes this algorithm very computationally efficient because the Modified Schmidt procedure only requires one data pass during the whole searching process. The selected features are compared to those obtained by other methods, through cross validation with support vector machines (SVMs).

The genetic study of titin has been notoriously difficult because of its size and complicated alternative splicing routes. Here, we have used zebrafish as an animal model to investigate the functions of individual titin isoforms. We identified 2 titin orthologs in zebrafish, ttna and ttnb , and annotated the full-length genomic sequences for both genes. We found that ttna , but not ttnb , is required for sarcomere assembly in the heart as well as the subsequent establishment of cardiac contractility. In fact, ttna is the earliest sarcomeric mRNA that is expressed in the heart, which makes it an early molecular marker for cardiomyocyte differentiation. Surprisingly, ttna is required for later steps of sarcomere assembly, including the assembly of Z-discs and A-bands, but not for early steps such as the assembly of Z-bodies and nonstriated myosin filaments. Reduction of individual titin isoforms in vivo using morpholino-modified antisense oligonucleotides indicated that (1) both N2B exon–containing and N2A exon–containing isoforms of ttna are required for sarcomere assembly in the heart; (2) N2A exon–containing isoforms of both ttna and ttnb are required for sarcomere assembly in the somites; and (3) the N2B exon–containing isoforms of ttnb are expressed later than other titin isoforms and are probably involved in modulating their expression; however, these isoforms of ttnb are not required for sarcomere assembly. Collectively, our results reveal distinct functions of different titin isoforms and suggest that various phenotypes in “titinopathies” may be attributable to the disruption of different titin isoforms.

Nitric oxide (NO) is a highly diffusible and short-lived physiological messenger. Despite its diffusible nature, NO modifies thiol groups of specific cysteine residues in target proteins and alters protein function via S-nitrosylation. Although intracellular S-nitrosylation is a specific posttranslational modification, the defined localization of an NO source (nitric oxide synthase, NOS) with protein S-nitrosylation has never been directly demonstrated. Endothelial NOS (eNOS) is localized mainly on the Golgi apparatus and in plasma membrane caveolae. Here, we show by using eNOS targeted to either the Golgi or the nucleus that S-nitrosylation is concentrated at the primary site of eNOS localization. Furthermore, localization of eNOS on the Golgi enhances overall Golgi protein S-nitrosylation, the specific S-nitrosylation of N -ethylmaleimide-sensitive factor and reduces the speed of protein transport from the endoplasmic reticulum to the plasma membrane in a reversible manner. These data indicate that local NOS action generates organelle-specific protein S-nitrosylation reactions that can regulate intracellular transport processes.

Abstract Pretreatment cytogenetics is a known predictor of outcome in hematologic malignancies. However, its usefulness in adult acute lymphoblastic leukemia (ALL) is generally limited to the presence of the Philadelphia (Ph) chromosome because of the low incidence of other recurrent abnormalities. We present centrally reviewed cytogenetic data from 1522 adult patients enrolled on the Medical Research Council (MRC) UKALLXII/Eastern Cooperative Oncology Group (ECOG) 2993 trial. The incidence and clinical associations for more than 20 specific chromosomal abnormalities are presented. Patients with a Ph chromosome, t(4;11)(q21;q23), t(8;14)(q24.1;q32), complex karyotype (5 or more chromosomal abnormalities), or low hypodiploidy/near triploidy (Ho-Tr) all had inferior rates of event-free and overall survival when compared with other patients. In contrast, patients with high hyperdiploidy or a del(9p) had a significantly improved outcome. Multivariate analysis demonstrated that the prognostic relevance of t(8;14), complex karyotype, and Ho-Tr was independent of sex, age, white cell count, and T-cell status among Ph-negative patients. The observation that Ho-Tr and, for the first time, karyotype complexity confer an increased risk of treatment failure demonstrates that cytogenetic subgroups other than the Ph chromosome can and should be used to risk stratify adults with ALL in future trials.