Effect of thyroid hormone and serum on the development of Na+, K+-adenosine triphosphatase and associated ion fluxes in cultures from rat brain

The effect of culture conditions, serum supplementation or chemically defined medium and the influence of thyroid hormone were studied on the development of the Na+, K+-adenosine triphosphatase (Na+,K+-ATPase) and on the intracellular content of K+ and Na+ ions in cultures which either were greatly enriched in a neuronal cell type, the cerebellar granule cells, or contained a mixed population of cells (brain reaggregates). Foetal rat brain reaggregates displayed lower Na+,K+-ATPase activity when cultured in chemically defined medium than in the presence of serum. Supplementation of the serum-free medium with thyroid hormone resulted in a rise in the Na+,K+-ATPase activity and [3H]ouabain binding to levels similar to those found in the cultures grown in the serum-containing medium. Thyroid hormone had no significant effect on the Mg2+-ATPase activity and on the intracellular content of Na+ and K+ ions. In the granule cell-enriched cerebellar surface cultures the Na+,K+-ATPase activity was lower when the cells were grown in chemically defined medium compared with the serum-containing medium, and the intracellular Na+ to K+ ratio was higher. Thyroid hormone had no effect on the Na+,K+-ATPase activity, [3H]ouabain binding or Mg2+-ATPase activity. The hormone also failed to influence ATPase activities in cerebellar astrocytes maintained in chemically defined medium. Although thyroid hormone had no effect on the Na+,K+-ATPase activity of cultured cerebellar granule cells, treatment with the hormone resulted in a decrease in the ratio of intracellular Na+ to K+ ion content. The effect of the hormone on the Na+,K+-pump activity in live cells was therefore tested by estimating ouabain-sensitive 86Rb uptake. This was regulated as in other cell types, by the rate of Na+ entry: the Na+-ionophore monensin trebled the rate of 86Rb uptake, which was also increased (+30-100%) by 10% foetal calf serum, the maximal response being obtained by about 20 min exposure to serum. The effect was completely blocked by the Na+/H+ exchange inhibitor amiloride. The factor(s) in the serum responsible for the regulation of the Na+,K+-pump were, however, not the thyroid hormones, which failed to affect 86Rb uptake. On the basis of comparing thyroid hormone effects on the different cultures studied it was concluded that not every type of neural cell is target of the hormone action during development.     

Electrophysiological properties of calcitonin-secreting cells derived from human medullary thyroid carcinoma

Primary cultures of human medullary thyroid carcinoma tissue were prepared from lymph node metastases in two patients. The parenchymal, cultured cells displayed positive immunocytochemical staining for CT, and the cells also released the hormone into the culture medium. The membrane potential and resistance of the CT-producing cells were 50.1 +/- 8.9 mV and 634 +/- 154 M omega (mean +/- SD, n = 46). TTX sensitive action potentials with maximum rate of rise up to 51 V s-1 were evoked by current injection in Na+-containing solution, whereas TTX insensitive action potentials with maximum rate of rise up to 9 V s-1 were generated in Na+-free solution. These action potentials were reversibly blocked by D-600. We conclude that the action potentials of the human MTC cells have both a Na+ and a Ca2+ component. Ejection of CA2+-free solution close to the cells caused membrane hyperpolarization associated with decreased membrane resistance. The reversal potential of this response was -66.2 +/- 10.9 mV (n = 10), indicating that a permeability increase to Cl- and/or K+ may be involved. We suggest that elevated plasma Ca2+ concentration in vivo may cause increased excitability due to membrane depolarization and resistance increase, thus leading to enhanced Ca2+ influx and hormone secretion.     

Changes in localization of ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase activity in human thyroid carcinoma cells

K+-dependent, ouabain-sensitive nitrophenyl phosphatase (K+-NPPase) activity, which reflects the terminal dephosphorylation step of (Na+ + K+)-ATPase action, was studied histochemically in human thyroid normal follicular cells and in human thyroid carcinoma cells, using a newly developed one-step lead citrate method. In normal thyroid follicular cells, reaction product for K+-NPPase activity was found on the lateral plasma membrane and not on either the apical or basal plasma membrane. In thyroid carcinoma cells, a large amount of reaction product was observed on the lateral plasma membrane and also on the apical and basal plasma membrane. Appropriate control experiments indicated that the deposition of reaction product was K+ dependent and ouabain sensitive. Although there was some overlap in the distribution of reaction products for K+-NPPase and Mg2+-ATPase, significant differences were consistently observed. The biochemical findings indicated that the K+-NPPase activity per milligram of DNA in thyroid carcinoma cells was approximately 10 times higher than that in normal thyroid cells, and that a significant positive correlation exists between K+-NPPase and (Na+ + K+)-ATPase activity. The physiologic and pathologic implications of this localization for tracing the route of active Na+ transport, which might participate in the transport of iodide ion in both human thyroid normal follicular cells and human thyroid carcinoma cells, are discussed.     

Alterations in monovalent cation transport in Sindbis virus-infected chick cells

Influx experiments using the potassium tracer 86Rb+ indicated that the activity of the Na+K+ ATPase, or sodium pump, was reduced 40-50% as a consequence of Sindbis virus infection of avian fibroblasts. The inhibition of this ouabain-sensitive, active transport system temporally correlated with a decrease in the intracellular K+ concentration and the termination of cellular protein synthesis. By contrast, the rate of influx facilitated by the furosemide-sensitive (Na+K+Cl-) cotransport system was only slightly depressed. Efflux experiments indicated that no alterations in the relative rate of nonspecific permeability or "leakage" of K+ could be detected in chick cells infected by Sindbis virus. The amount of [3H]ouabain bound to Sindbis virus-infected cells paralleled the reduction in Na+K+ ATPase activity. These binding studies revealed no difference in the number of Na+ pump sites. The Km of ouabain binding, however, increased approximately 3.5-fold in the virus-infected cells. No change in the apparent affinity of the Na+ pump for K+ could be detected, yet the Vmax for ouabain-sensitive K+ transport was decreased. These experiments suggest that a reduction in Na+K+ ATPase turnover results in the altered intracellular monovalent cation levels found in Sindbis virus-infected chick cells.     

Acetyl-L-carnitine induces a sustained potentiation of the afterhyperpolarization

Acetyl-L-carnitine is known to improve many aspects of the neural activity even if its exact role in neurotransmission is still unknown. This study investigates the effects of acetyl-L-carnitine in T segmental sensory neurons of the leech Hirudo medicinalis. These neurons are involved in some forms of neural plasticity associated with learning processes. Their physiological firing is accompanied by a large afterhyperpolarization that is mainly due to the Na+/K+ ATPase activity and partially to a Ca2+ -dependent K+ current. A clear-cut hyperpolarization and a significant increase of the afterhyperpolarization have been recorded in T neurons of leeches injected with 2 mM acetyl-L-carnitine some days before. Acute treatments of 50 microM acetyl-L-carnitine induced similar effects in T cells of naive animals. In the presence of apamin, a pharmacological blocker of Ca2+ -dependent K+ channel, acetyl-L-carnitine still enhanced the residual afterhyperpolarization, suggesting an effect of the drug on the Na+/K+ATPase. Acetyl-L-carnitine also increased the hyperpolarization induced by intracellular injection of Na+ ions. Therefore, acetyl-L-carnitine seems to be able to exert a positive sustained effect on the Na+/K+ ATPase activity in leech T sensory neurons. Moreover, in these cells, widely arborized, the afterhyperpolarization seems to play an important role in determining the action potential transmission at neuritic bifurcations. A computational model of a T cell has been previously developed considering detailed data for geometry and the modulation of the pump current. Herein, we showed that to a larger afterhyperpolarization, due to the acetyl-L-carnitine-induced effects, corresponds a decrement in the number of action potentials reaching synaptic terminals.     

Sodium-potassium ATPase 1 subunit is a molecular partner of Wolframin, an endoplasmic reticulum protein involved in ER stress

Wolfram syndrome, an autosomal recessive disorder characterized by diabetes mellitus and optic atrophy, is caused by mutations in the WFS1 gene encoding an endoplasmic reticulum (ER) membrane protein, Wolframin. Although its precise functions are unknown, Wolframin deficiency increases ER stress, impairs cell cycle progression and affects calcium homeostasis. To gain further insight into its function and identify molecular partners, we used the WFS1-C-terminal domain as bait in a yeast two-hybrid screen with a human brain cDNA library. Na+/K+ ATPase beta1 subunit was identified as an interacting clone. We mapped the interaction to the WFS1 C-terminal and transmembrane domains, but not the N-terminal domain. Our mapping data suggest that the interaction most likely occurs in the ER. We confirmed the interaction by co-immunoprecipitation in mammalian cells and with endogenous proteins in JEG3 placental cells, neuroblastoma SKNAS and pancreatic MIN6 beta cells. Na+/K+ ATPase beta1 subunit expression was reduced in plasma membrane fractions of human WFS1 mutant fibroblasts and WFS1 knockdown MIN6 pancreatic beta-cells compared with wild-type cells; Na+/K+ ATPase alpha1 subunit expression was also reduced in WFS-depleted MIN6 beta cells. Induction of ER stress in wild-type cells only partly accounted for the reduced Na+/K+ ATPase beta1 subunit expression observed. We conclude that the interaction may be important for Na+/K+ ATPase beta1 subunit maturation; loss of this interaction may contribute to the pathology seen in Wolfram syndrome via reductions in sodium pump alpha1 and beta1 subunit expression in pancreatic beta-cells.     

Effect of insulin upon membrane-bound (Na+ + K+)-ATPase extracted from frog skeletal muscle.

1. Insulin stimulates the activity of membrane-bound ATPase isolated from frog skeletal muscle and from rat brain. The increase in activity of the membrane-bound ATPase system isolated from frog ranged from 9-8 to 53% at concentrations of Na+ (25 mM), K+ (10 mM), and ATP (2 mM) similar to those in in vivo experiments conducted previously (Moore, 1973). The increased activity of the membrane-bound ATPase is, therefore, at least as great as the insulin-induced increase in Na efflux (10-38%) from intact cells (Moore, 1973). If the concentration of Na+ is lowered to 4 mM and that of ATP lowered to 0-5 mM albumin, and 10(6) M, the increase in ouabain-inhibitable ATPase activity can reach as high as 400%. 2. Ouabain, at a concentration (10(-3) M) sufficient to inhibit stimulation of the frog ATPase by increasing Na from 4 to 25 mM, completely blocked the stimulation of ATPase activity due to insulin. 3. At 2 mM-ATP, 100 mM-Na+, and 20 mM-K+, conditions which maximally activate the (Na+ + K+)-ATPase, insulin did not increase the ATPase, activity. Stimulation was consistently seen at 10 mM-K+, 0-5 mM-ATP, and either 4 mM or 25 mM-Na+. 4. The finding that insulin does not stimulate the ATPase activity in conditions in which the (Na+ + K+)-ATPase component is maximally activated and especially the fact that ouabain can reproducibly inhibit insulin stimulation of the membrane-bound ATPase activity strongly suggest that interaction of insulin with its receptor upon the plasma membrane somehow stimulates the (Na+ + K+)-ATPase system (ouabain sensitive; ATP phosphohydrolase, EC (3.6.1.3). These results are consistent with previous studies of the effect of insulin upon Na efflux from intact cells (Moore, 1973) and support the previous conclusion that the component of Na efflux stimulated by insulin is active. The evidence suggests that insulin probably does not affect Vmax of the (Na+ + K+)-ATPase system, but may increase the affinity of the enzyme system to one or more effectors, most likely Na+, ATP, and perhaps K+. 5. Oxidized glutathione (2-7 X 10(-6) M), 10(-6) M, 10(-7) M, and 10(-8) M cyclic AMP did not affect the ATPase activity 10(-6)Malbumin, and . 6. The results are consistent with the view that the Na pump, (Na+ + K+)-ATPase, is intimately involved with the physiological action of insulin and may be transducer between the binding of insulin to its receptor on the plasma membrane and the cellular actions of insulin.     

Adhesion of K88ab to guinea pig erythrocytes: effect on membrane enzyme activities.

Nontoxigenic Escherichia coli strains bearing K88 fimbriae have been associated with diarrhea in piglets. We have used guinea pig erythrocytes as a model of the host cell to study the cellular alterations after adherence of purified K88ab fimbriae. Although Mg2+-dependent ATPase was inhibited (up to 61%), Na/K ATPase was not. Metabolic enzymes were not significantly affected.     

Mutagenic potency at the Na+/K+ ATPase locus correlates with cycle-dependent killing of 10T1/2 cells

Perturbation of DNA replication by chemical-DNA adducts produced by exposure to mutagenic/carcinogenic chemicals results in mutagenic or cytotoxic damage in the DNA. Demonstration of a correlation between cell cycle dependency of cytotoxicity and point mutation at the Na+/K+ ATPase gene could suggest that the two consequences of chemical exposure are caused by the same damage in the template DNA and that both are mediated through DNA replication-associated mechanisms. N-methyl-N'-nitro-N-nitrosoguanidine, N-ethyl-N'-nitro-N-nitrosoguanidine, 4-nitroquinoline-1-oxide, and benzo(a)pyrene-trans-7,8-dihydrodiol-9,10-epoxide demonstrated cell cycle-related patterns of cytotoxicity in 10T1/2 cells, with maximal cell killing produced by exposure in early S phase, and were highly efficient mutagens of the Na+/K+ ATPase gene relative to their cytotoxic potential. In contrast, methyl methanesulfonate and N-acetoxy-N-2-fluorenylacetamide were maximally cytotoxic in cell populations exposed in early G1 phase and were weak mutagens of the Na+/K+ ATPase gene at comparable levels of cytotoxicity. These data suggest that mutagenic/carcinogenic chemicals that are effective at producing mutations by misreplication kill cells by a related mechanism that may be associated with the perturbation of DNA replication.     

A circulating inhibitor of the platelet Na+,K+ adenosine triphosphatase (ATPase) enzyme in allergy.

Previous investigations have documented a reduced activity of the sodium-potassium-stimulated adenosine triphosphatase enzyme (Na+,K+ ATPase) in platelet membranes of allergic subjects. The purpose of this study was to determine if the reduced Na+,K+ ATPase activity was due to an enzyme inhibitor. Na+,K+ ATPase activity of a particulate fraction of sonicated platelets was determined by spectrophotometry in asymptomatic adults with and without allergy. The Na+,K+ ATPase level (mean, nanomoles per microgram of protein per minute; +/- STD) of allergic subjects (0.9 +/- 1.3) was lower (p less than 0.001) than that of nonallergic subjects (3.9 +/- 1.6). In contrast, when the same platelet fractions were frozen before assay, Na+,K+ ATPase was higher (p less than 0.005) in allergic subjects (6.0 +/- 1.4) than in nonallergic subjects (3.6 +/- 2.0). An inhibitor of canine kidney Na+,K+ ATPase was detected in the buffer in which these platelet fractions were frozen, allergic subjects (0.5% +/- 0.4% inhibition per microgram of protein) compared to nonallergic subjects (0.04% +/- 0.08%; p less than 0.005). The level of inhibition correlated positively with the postfreezing increase in platelet membrane Na+,K+ ATPase, suggesting a freezing-induced displacement of an inhibitor from the membrane. Plasma from these same subjects inhibited Na+,K+ ATPase activity of normal platelets, allergic subjects (70% +/- 31% inhibition) compared to nonallergic subjects (13% +/- 16%; p less than 0.001). These data suggest that the transport-enzyme defect observed in platelets from allergic subjects was due to a circulating Na+,K+ ATPase inhibitor. In vivo Na+,K+ ATPase inhibition in allergy could have profound effects on intracellular cation concentrations and broad implications for pathogenesis.     

Influence of the indigenous gastrointestinal microbial flora on duodenal Mg2+ -dependent and (Na+ + K+) -stimulated adenosine triphosphatase activities in mice.

Mg-dependent and (Na+ + K+)-stimulated adenosine triphosphatase (ATP-ase) activities were assayed in butanol extracts of duodenal tissue from germ-free, specific-pathogen-free, and ex-germfree mice associated with an indigenous microflora from specific-pathogen-free mice. In the germfree mice the levels of both ATPase activities were significantly higher than the levels in specific-pathogen-free mice. By contrast, the ex-germfree animals colonized by an entire indigenous microflora, the values fell to levels close to those for specific-pathogen-free animals. (Na+ + K+)-stimulated ATPase was not inhibited by ouabain in extracts from any of the three kinds of mice. These findings show that the indigenous microbial flora influences the intestinal ATPase activity of mice and, because of the connection between (Na+ + K+-stimulate ATPase and active transpoort, undoubtedly affects the process of absorption in the intestinal tract.     

Magnesium-dependent adenosine triphosphatase as a marker enzyme for the plasma membrane of human polymorphonuclear leukocytes.

The adenosine triphosphatase (ATPase) activities of human polymorphonuclear leukocytes (PMNL) were studied with an assay that monitored the release of 32P-labeled inorganic pyrophosphate (32P1) from gamma-[32P]adenosine 5'-triphosphate (ATP). In cell homogenates, (Na+ + K+)-sensitive, ouabain-inhibitable ATPase comprised an insignificant fraction of the total ATPase activity. Additions of p-nitrophenyl phosphate and beta-glycerophosphate (substrates for nonspecific acid and alkaline phosphatases) and of tartrate (inhibitor of acid phosphatase) gave no indication of inhibition. This suggested that the assay was relatively specific for ATP hydrolysis. The activity was found to have a pH optimum of 8.7 and a Km for ATP of 0.6 mM. There was an absolute requirement for Mg2+, with other divalent cations substituting less efficiently. When the Mg2+-dependent ATPase activity of intact cells was compared with that in homogenized cells, no significant difference was observed. The activity in intact cells was linear with respect to incubation time up to at least l0 min. Trypan blue staining and lactate dehydrogenase assays revealed that greater than 92% of the PMNL remained intact and viable during the assay. No soluble ATPase was released from the cells under assay conditions. In following the distribution of gamma[32P]ATP and 32P2 counts became cell associated. Since the experimental evidence supports the observation that PMNL remain intact and viable and that ATP does not penetrate the cell under assay conditions, it is proposed that greater than 90% of the Mg2+-dependent ATPase of the human PMNL is associated with a plasma membrnae enzyme. This would qualify the enzyme for the role of a plasma membrane marker for future fractionation and isolation attempts.     

Na+,K+-ATPase activity is inhibited in cultured intestinal epithelial cells by endotoxin or nitric oxide

Na+K+-ATPase is an important enzyme serving vital functions in various mammalian tissues, including the intestine. We have previously documented that endotoxin (LPS) and nitric oxide (NO) can induce enterocyte injury in vitro. To examine whether alterations Na+,K+-ATPase activity might be involved in LPS- or NO-induced enterocyte dysfunction, we carried out four series of experiments. The first set of experiments documented that LPS decreases IEC-6 Na+,K+-ATPase activity at concentrations as low as 0.10 microg/ml. The second set of experiments tested whether exposure of IEC-6 cells to the exogenous NO donor, S-Nitroso-N-acetylpenicillamine (SNAP), would decrease IEC-6 Na+,K+-ATPase activity. The results of these experiments documented that SNAP significantly decreased IEC-6 Na+,K+-ATPase activity in a dose-dependent fashion at a threshold inhibitory concentration of 0.1 mM, and there was an inverse correlation between Na+,K+-ATPase activity and NO concentrations in the medium. Since enterocytes contain iNOS, and LPS can increase iNOS activity, the third set of experiments examined the relationship between LPS-induced inhibition of Na+),K+-ATPase activity and NO production by the IEC-6 cells. These results showed that LPS increased IEC-6 NO production in both a dose- and time-dependent fashion and an inverse correlation existed between LPS-induced NO production and decreased Na+,K+-ATPase activity. Addition of the NOS inhibitor, L-NNA, prevented the LPS-induced decrease in Na+,K+ATPase activity, suggesting that NO is involved in the decrease of Na+,K+-ATPase activity observed in the IEC-6 cells incubated with LPS. One mechanism by which the increased NO concentrations could have contributed to the decrease in Na+,K+ATPase activity, after the addition of LPS or SNAP, is via the production of peroxynitrite during the reaction of NO with superoxide. This notion was supported by studies showing that SNAP- and LPS-induced decreases in IEC-6 Na+,K+-ATPase activity could be blocked by adding superoxide dismutase to the medium. The last set of experiments tested whether the inhibition of Na+,K+-ATPase activity with the specific Na+,K+-ATPase inhibitor ouabain would increase the permeability of an IEC-6 monolayer. IEC-6 monolayer permeability was increased by ouabain, but only at a high concentration. In conclusion, these studies indicate that LPS or the NO donor, SNAP, inhibit Na+,K+-ATPase activity and this inhibition is at least partly related to peroxynitrite production. These studies also suggest that LPS-induced NO production by the IEC-6 cells decreases IEC-6 Na+,K+-ATPase activity in an autocrine fashion.     

Lamin B1 depletion in senescent cells triggers large-scale changes in gene expression and the chromatin landscape

Senescence is a stable proliferation arrest, associated with an altered secretory pathway, thought to promote tumor suppression and tissue aging. While chromatin regulation and lamin B1 down-regulation have been implicated as senescence effectors, functional interactions between them are poorly understood. We compared genome-wide Lys4 trimethylation on histone H3 (H3K4me3) and H3K27me3 distributions between proliferating and senescent human cells and found dramatic differences in senescence, including large-scale domains of H3K4me3- and H3K27me3-enriched “mesas” and H3K27me3-depleted “canyons.” Mesas form at lamin B1-associated domains (LADs) in replicative senescence and oncogene-induced senescence and overlap DNA hypomethylation regions in cancer, suggesting that pre-malignant senescent chromatin changes foreshadow epigenetic cancer changes. Hutchinson-Gilford progeria syndrome fibroblasts (mutant lamin A) also show evidence of H3K4me3 mesas, suggesting a link between premature chromatin changes and accelerated cell senescence. Canyons mostly form between LADs and are enriched in genes and enhancers. H3K27me3 loss is correlated with up-regulation of key senescence genes, indicating a link between global chromatin changes and local gene expression regulation. Lamin B1 reduction in proliferating cells triggers senescence and formation of mesas and canyons. Our data illustrate profound chromatin reorganization during senescence and suggest that lamin B1 down-regulation in senescence is a key trigger of global and local chromatin changes that impact gene expression, aging, and cancer.     

Mini-review: Cell surface receptor for thyroid hormone and nongenomic regulation of ion fluxes in excitable cells

Thyroid hormone has been shown experimentally to affect cellular ion fluxes. For example, thyroid hormone-induced modulation has been described of cellular sodium current (I_Na), inward rectifying potassium current (IKir) and sodium pump (Na, K-ATPase) and of calcium pump (Ca²⁺-ATPase) activities. Certain of these actions appear to reflect nongenomic mechanisms of hormone action that are initiated at the plasma membrane receptor for iodothyronines described on integrin αvβ3. One such action is the recent demonstration of enhancement by the hormone of I_Na in neurons. Nongenomic actions of thyroid hormone initiated at the plasma membrane may be specifically inhibited by tetraiodothyroacetic acid (tetrac), a deaminated thyroid hormone analogue.Important behavioral changes are associated with clinical states of excessive or deficient thyroid function. The molecular basis for these changes has not been established. It is proposed that nongenomic actions of thyroid hormone in neurons—such as that on sodium current—underlie certain of these behaviors. The contribution of such nongenomic actions of the hormone to animal behavioral paradigms possibly relevant to thyroid hormone actions in human subjects may be tested in vivo with tetrac.     

兔角膜内皮细胞Na~+-K~+-ATP酶的电镜酶细胞化学观察

目的　探讨兔角膜内皮细胞上Na+ K+ ATP酶的分布。方法　本文采用电镜酶组化方法对兔角膜内皮细胞Na+ K+ ATP酶的分布进行观察。结果　角膜内皮细胞面向前房部分的细胞膜的Na+ K+ ATP酶活性较高。结论　内皮细胞活性的检测有利于角膜保存的研究     

Extracellular signals in young and aging breast epithelial cells and possible connections to age-associated breast cancer development

Aging of human breast tissue is accompanied by certain structural and functional variations and several studies suggest a possible contribution of these changes to an aging-related breast cancer development. At the cellular level, aging of human mammary epithelial cells is associated with significant morphological and functional alterations such as an increased cell size and a reduced proliferation. Cellular senescence of HMEC cannot be explained by a single mechanism but represents an interaction of numerous extra- and intracellular events and the complexity of such orchestrating pathways is still hardly understood. Besides the contribution of reactive oxygen species and telomere dysfunction to aging, it is the aim of this mini-review, to compare distinct changes to extracellular signals by certain matrix metalloproteinases including MMP-7 and associated growth factor pathways mediated by HB-EGF activation in young and aging HMEC. Such changes can alter hormone receptor levels within aged HMEC, induce tissue fibrosis and promote epithelial-to-mesenchymal transition as a potential prerequisite for breast cancer development. Moreover, an accumulation of aging cells during the normal life span of the breast tissue may also substantially effect and interact with adjacent neighboring populations in the local microenvironment to provide optimized growth conditions which would also support neoplastic cells.     

Some properties of the adenosine triphosphatase associated with herpes simplex virus and nuclear membrane of host cells.

An enzyme capable to split adenosine triphosphate (ATP) was shown to be firmly associated with mature herpes simplex virus particles purified from infected rabbit lung (ZP) cells. The enzyme localized in the viral envelope was markedly activated by bivalent cations, to the largest degree by Mg2+ at a pH optimum of 7.8--8.0. Na+ and K+ ions neither separately nor together showed any activating effect. Enzyme activity was not sensitive to the action of ouabain. No adenosine diphosphatase (ADPase) and adenosine monophosphatase (AMPase) activities were observed. ATPase activity was competitively inhibited by ADP. AMP and inorganic phosphate were without effect. The ATPase of nuclear membranes isolated from ZP cells exhibited similar properties but behaved differently to the action of sodium dithionite, dinitrophenol, oligomycin and gramicidin, as well as on heat inactivation. The origin of the virus enzyme is discussed.     

Promotion of the induction of cell pluripotency through metabolic remodeling by thyroid hormone triiodothyronine-activated PI3K/AKT signal pathway

Generation of induced pluripotent stem cells (iPSCs) from somatic cells by defined factors is a mechanism-unknown, yet extremely time-consuming process. Inefficient reprogramming leads to prolonged periods of in?vitro iPSC selection, resulting in subtle genetic and epigenetic abnormalities. To facilitate pluripotent reprogramming, we have identified the thyroid hormone triiodothyronine (T3) as an endogenous factor that can enhance reprogramming of human dermal fibroblasts (HDF) and umbilical cord mesenchymal stem cells (UCMSC). This potentiation of iPSC induction is associated with metabolic remodeling activity, including upregulation of key glycolytic genes, an increase in cell proliferation, and the induction of mesenchymal-epithelial transition (MET). We further identify the activation of the PI3K/AKT signal pathway by T3 as an underlying mechanism for the enhanced conversion to cell pluripotency in this model. These studies demonstrate that T3 enhances metabolic remodeling of donor cells in potentiating cell reprogramming.