Chronic Metabolic Stress Research Articles

Efficient tumorigenesis by mutation-induced failure to terminate microRNA-mediated adaptive hyperplasia

Seven current contending cancer theories consider different sets of critical events as sufficient for tumorigenesis. These theories, most recently the microRNA dysregulation (MRD) theory, have overlapping attributes and extensive empirical support, but also some discrepancies, and some do not address both benign and malignant tumorigenesis. By definition, the most efficient tumorigenic pathways will dominate under conditions that selectively activate those pathways. The MRD theory provides a mechanistic basis to combine elements of the current theories into a new hypothesis that: (i) tumors arise most efficiently under stress that induces and sustains either protective or regenerative states of adaptive hyperplasia (AH) that normally are epigenetically maintained unless terminated; and (ii) if dysregulated by a somatic mutation that prevents normal termination, these two AH states can generate benign and malignant tumors, respectively. This hypothesis, but not multistage cancer theory, predicts that key participating AH-stem-cell populations expand markedly when triggered by stress, particularly chronic metabolic or oxidative stress, mechanical irritation, toxic exposure, wounding, inflammation, and/or infection. This hypothesis predicts that microRNA expression patterns in benign vs. malignant tumor tissue will correlate best with those governing protective vs. regenerative AH in that tissue, and that tumors arise most efficiently inmutagen-exposed stem cells that either happen to be in, or incidentally later become recruited into, an AH state.

Imaging Functional Beta Cell Mass: Can we See Islets Clearly Now?

Beta cell mass is dynamic, and changes during neonatal growth and development, during pregnancy, and in response to chronic metabolic stress, such as obesity and diabetes. Molecular imaging techniques can be used to provide real-time readouts on subclinical changes in beta cell mass, and, in the process, enhance our understanding of the molecular processes that govern its regulation. The strategy of engineering beta cells to express endogenous contrast for imaging by fluorescence, bioluminescence and positron emission tomography (PET) has been especially useful in developing novel insights into the timing of the decline in functional beta cell mass during the progression of diabetes. There have also been some recent exciting developments in the use of MRI in detecting beta cell function by tagging the movement of cations across the cell membrane in response to glucose. While such imaging strategies may not be immediately translated to the clinic, they have provided the opportunity to directly visualize the islet development and formation as well as the subclinical declines in beta cell mass that precede the onset of overt diabetes. This review will discuss the impact of transgenic mouse models and MR imaging of cation flux on the field of beta cell imaging.

Metabolic Remodeling in the Hypertrophic Heart

When it comes to fuel for energy production, it is commonly accepted that the heart is an omnivore, capable of oxidizing a wide range of carbon substrates, and that this metabolic plasticity is necessary to maintain a high and variable workload in the midst of an ever-changing hormonal and nutritional state.1 It is also widely appreciated that shifts in substrate utilization in the heart occur in response to chronic metabolic and hemodynamic stresses.2–5 What is less clear, however, is whether such chronic metabolic shifts should be considered a cause or consequence in the pathogenesis of contractile dysfunction. In the case of diabetes mellitus, the heart exhibits an increased dependence on fatty acids for oxidative energy production, and this increase in lipid metabolism has been proposed to significantly contribute to the etiology of impaired cardiac function.6 Similarly, increased rates of fatty acid oxidation immediately after an ischemic event have been implicated in exacerbation of reperfusion injury.7 However, in both of these pathophysiological settings, compelling data to support a direct causal relationship between contractile abnormalities and metabolic dysregulation remain elusive. Article, see p 728 In response to pressure overload–induced cardiac hypertrophy, the heart reverts more toward a fetal-like metabolic profile, indicative of a decrease in fatty acid oxidation (concomitant with an increased reliance on carbohydrates for oxidative energy metabolism).3 It has been suggested that this substrate shift, which is associated with reactivation of other fetal-like hallmarks (eg, myosin heavy chain isoform switching), contributes to the progression to overt contractile failure.8 Dietary, pharmacological, and genetic strategies have been used in attempts to provide insight regarding the impact that substrate shifts have on pressure overload–induced remodeling; however, mixed results have been reported. …

Vascular Klotho Deficiency Potentiates the Development of Human Artery Calcification and Mediates Resistance to Fibroblast Growth Factor 23

Klotho is known to function as a cofactor for the phosphatonin, fibroblast growth factor (FGF)-23 at the kidney. FGF-23 levels rise in chronic kidney disease (CKD) despite progression of accelerated vascular calcification. There are currently conflicting data on whether FGF-23 may exhibit direct vasculoprotective effects in CKD. In this study, we describe for the first time endogenous Klotho expression in human arteries and human aortic smooth muscle cells. We show that CKD is a state of vascular Klotho deficiency promoted by chronic circulating stress factors, including proinflammatory, uremic, and disordered metabolic conditions. Mechanistic studies demonstrated that Klotho knockdown potentiated the development of accelerated calcification through a Runx2 and myocardin-serum response factor-dependent pathway. Klotho knockdown studies further revealed that vascular cells are a Klotho-dependent target tissue for FGF-23. FGF-23 mediated cellular activation of p-ERK, p-AKT, and cellular proliferative effects, which were abrogated following Klotho knockdown. We next showed that vascular Klotho deficiency driven by procalcific stressors could be restored by vitamin D receptor activators, in vitro and further confirmed using human arterial organ cultures from CKD patients, in vivo. Furthermore, restoration of suppressed Klotho expression by vitamin D receptor activators conferred human aortic smooth muscle cells responsive to FGF-23 signaling and unmasked potential anticalcific effects. Chronic metabolic stress factors found in CKD promote vascular Klotho deficiency. Mechanistic studies revealed a bifunctional role for local vascular Klotho, first, as an endogenous inhibitor of vascular calcification and, second, as a cofactor required for vascular FGF-23 signaling. Furthermore, vitamin D receptor activators can restore Klotho expression and unmask FGF-23 anticalcific effects.

Role of carnitine in the regulation of glucose homeostasis and insulin sensitivity: evidence from in vivo and in vitro studies with carnitine supplementation and carnitine deficiency

Although carnitine is best known for its role in the import of long-chain fatty acids (acyl groups) into the mitochondrial matrix for subsequent β-oxidation, carnitine is also necessary for the efflux of acyl groups out of the mitochondria. Since intracellular accumulation of acyl-CoA derivatives has been implicated in the development of insulin resistance, carnitine supplementation has gained attention as a tool for the treatment of insulin resistance. More recent studies even point toward a causative role for carnitine insufficiency in developing insulin resistance during states of chronic metabolic stress, such as obesity, which can be reversed by carnitine supplementation. The present review provides an overview about data from both animal and human studies reporting effects of either carnitine supplementation or carnitine deficiency on parameters of glucose homeostasis and insulin sensitivity in order to establish the less well-recognized role of carnitine in regulating glucose homeostasis. Carnitine supplementation studies in both humans and animals demonstrate an improvement of glucose tolerance, in particular during insulin-resistant states. In contrast, less consistent results are available from animal studies investigating the association between carnitine deficiency and glucose intolerance. The majority of studies dealing with this question could either find no association or even reported that carnitine deficiency lowers blood glucose and improves insulin sensitivity. In view of the abovementioned beneficial effect of carnitine supplementation on glucose tolerance during insulin-resistant states, carnitine supplementation might be an effective tool for improvement of glucose utilization in obese type 2 diabetic patients. However, further studies are necessary to explain the conflicting observations from studies dealing with carnitine deficiency.

Factors regulating fat oxidation in human skeletal muscle

In modern societies, oversupply of calories leads to obesity and chronic metabolic stress, which may lead to development of disease. Oversupply of calories is often associated with elevated plasma lipid concentrations and accumulation of lipids in skeletal muscle leading to decreased insulin sensitivity. Consequently, enhanced fat oxidation might be beneficial in counteracting lipid accumulation. Exercise is the most effective way to increase fat oxidation, because it increases metabolic rate. Lipid metabolism can also be altered by dietary manipulations. Thus, a fat rich diet leads to increased potential for fat oxidation by increasing the content of several of the proteins in the fat oxidative pathway. The regulation of both exercise and diet induced lipid oxidation will be discussed in this review.

Health status among prehistoric Eskimos from Point Hope, Alaska

Using the protocol outlined in The Backbone of History: Health and Nutrition in the Western Hemisphere (BBH) (Steckel and Rose. 2002a. The backbone of history: health and nutrition in the Western Hemisphere. Cambridge: Cambridge University Press), this project compares the Mark I Health Index (MIHI) scores of the Ipiutak (n = 76; 100BCE-500CE) and Tigara (n = 298; 1200-1700CE), two samples of North American Arctic Eskimos excavated from Point Hope, Alaska. Macroscopic examination of skeletal remains for evidence of anemia, linear enamel hypoplasias (LEH), infection, trauma, dental health, and degenerative joint disease (DJD) was conducted to assess differences in health status resulting from a major economic shift at Point Hope. These data demonstrate that despite differences in settlement pattern, economic system, and dietary composition, the MIHI scores for the Ipiutak (82.1) and Tigara (84.6) are essentially equal. However, their component scores differ considerably. The Ipiutak component scores are suggestive of increased prevalence of chronic metabolic and biomechanical stresses, represented by high prevalence of nonspecific infection and high frequencies of DJD in the hip/knee, thoracic vertebrae, and wrists. The Tigara experienced more acute stress, evidenced by higher prevalence of LEH and trauma. Comparison of overall health index scores with those published in BBH shows the MIHI score for the Ipiutak and Tigara falling just above the average for sites in the Western Hemisphere, adding support to the argument that the human capacity for cultural amelioration of environmental hardships is quite significant.

Oxidative Damage Compromises Energy Metabolism in the Axonal Degeneration Mouse Model of X-Adrenoleukodystrophy

Chronic metabolic impairment and oxidative stress are associated with the pathogenesis of axonal dysfunction in a growing number of neurodegenerative conditions. To investigate the intertwining of both noxious factors, we have chosen the mouse model of adrenoleukodystrophy (X-ALD), which exhibits axonal degeneration in spinal cords and motor disability. The disease is caused by loss of function of the ABCD1 transporter, involved in the import and degradation of very long-chain fatty acids (VLCFA) in peroxisomes. Oxidative stress due to VLCFA excess appears early in the neurodegenerative cascade. In this study, we demonstrate by redox proteomics that oxidative damage to proteins specifically affects five key enzymes of glycolysis and TCA (Tricarboxylic acid) cycle in spinal cords of Abcd1(-) mice and pyruvate kinase in human X-ALD fibroblasts. We also show that NADH and ATP levels are significantly diminished in these samples, together with decrease of pyruvate kinase activities and GSH levels, and increase of NADPH. Treating Abcd1(-) mice with the antioxidants N-acetylcysteine and α-lipoic acid (LA) prevents protein oxidation; preserves NADH, NADPH, ATP, and GSH levels; and normalizes pyruvate kinase activity, which implies that oxidative stress provoked by VLCFA results in bioenergetic dysfunction, at a presymptomatic stage. Our results provide mechanistic insight into the beneficial effects of antioxidants and enhance the rationale for translation into clinical trials for X-adrenoleukodystrophy.

Athletic amenorrhea: energy deficit or psychogenic challenge?

Athletic women are at risk for developing ovulatory dysfunction, which presents variably as menstrual irregularity or absence. Initially characterized as an isolated disruption of hypothalamic gonadotropin-releasing hormone (GnRH) release, athletic amenorrhea, a form of hypogonadotropic hypogonadism, is invariably accompanied by additional neuroendocrine aberrations, including activation of adrenal and suppression of thyroidal axes. Exercise may elicit intermittent or chronic metabolic stress owing to increased energy expenditure and/or insufficient or imbalanced nutrient intake. In addition, athletic activities are motivated by or serve as psychogenic stressors. Prior studies dichotomized stressors as metabolic or psychogenic. Not only is this a false dichotomy because all stressors have both a metabolic and a psychogenic component, but also stressors act synergistically rather than in isolation to compromise GnRH drive and endocrine homeostasis. To ameliorate reproductive and endocrine consequences of stress, then, requires identification and amelioration of all relevant stressors. Formal psychosocial support helps individuals to develop better coping strategies and make appropriate lifestyle changes. Our research has shown that cognitive behavior therapy restores reproductive and endocrine balance.

Acute Modulation of Sugar Transport in Brain Capillary Endothelial Cell Cultures during Activation of the Metabolic Stress Pathway

GLUT1-catalyzed equilibrative sugar transport across the mammalian blood-brain barrier is stimulated during acute and chronic metabolic stress; however, the mechanism of acute transport regulation is unknown. We have examined acute sugar transport regulation in the murine brain microvasculature endothelial cell line bEnd.3. Acute cellular metabolic stress was induced by glucose depletion, by potassium cyanide, or by carbonyl cyanide p-trifluoromethoxyphenylhydrazone, which reduce or deplete intracellular ATP within 15 min. This results in a 1.7-7-fold increase in V(max) for zero-trans 3-O-methylglucose uptake (sugar uptake into sugar-free cells) and a 3-10-fold increase in V(max) for equilibrium exchange transport (intracellular [sugar] = extracellular [sugar]). GLUT1, GLUT8, and GLUT9 mRNAs are detected in bEnd.3 cells where GLUT1 mRNA levels are 33-fold greater than levels of GLUT8 or GLUT9 mRNA. Neither GLUT1 mRNA nor total protein levels are affected by acute metabolic stress. Cell surface biotinylation reveals that plasma membrane GLUT1 levels are increased 2-3-fold by metabolic depletion, although cell surface Na(+),K(+)-ATPase levels remain unaffected by ATP depletion. Treatment with the AMP-activated kinase agonist, AICAR, increases V(max) for net 3-O-methylglucose uptake by 2-fold. Glucose depletion and treatment with potassium cyanide, carbonyl cyanide p-trifluoromethoxyphenylhydrazone, and AICAR also increase AMP-dependent kinase phosphorylation in bEnd.3 cells. These results suggest that metabolic stress rapidly stimulates blood-brain barrier endothelial cell sugar transport by acute up-regulation of plasma membrane GLUT1 levels, possibly involving AMP-activated kinase activity.

A review of the influence of low ambient calcium concentrations on freshwater daphniids, gammarids, and crayfish

The widespread decline in aqueous calcium (Ca) is emerging as a newly recognised stressor in freshwater ecosystems in regions with historically high acid deposition, especially when coupled with multiple logging cycles. Currently, Ca and other base cations are being depleted in the soils of acid-sensitive watersheds of eastern North America and western Europe, resulting in falling Ca levels in streams and lakes. Freshwater crustaceans have high Ca demands due to their heavily calcified exoskeleton and regular moult cycle. Because they rely on Ca in the external environment for the majority of their Ca uptake, aquatic crustaceans are restricted to waters at or above the Ca concentration needed to satisfy their demands. The zoogeographic distributions along ambient Ca gradients, and Ca requirements and metabolism of three groups of freshwater crustaceans — daphniids, crayfish and gammarids — have been relatively well studied. Here we have four objectives: (1) We briefly review biological features of the above three taxa that relate to Ca metabolism. (2) We review the literature regarding minimum Ca thresholds permitting survival, Ca saturation points, and the individual and population scale costs of existence at suboptimal Ca concentrations. (3) Using Daphnia as an example, we explore the ecological consequences of falling environmental Ca concentrations. (4) We identify gaps and weaknesses in the literature that may inhibit the development of environmental risk assessments for Ca decline for these three crustacean groups. We conclude that crayfish are especially vulnerable to ongoing Ca decline, and that all three taxa are probably living under suboptimal conditions in areas of Ca decline, and are therefore likely under chronic metabolic stress.

Mitochondrial Complex II Dysfunction Can Contribute Significantly to Genomic Instability after Exposure to Ionizing Radiation

Ionizing radiation induces chronic metabolic oxidative stress and a mutator phenotype in hamster fibroblasts that is mediated by H(2)O(2), but the intracellular source of H(2)O(2) is not well defined. To determine the role of mitochondria in the radiation-induced mutator phenotype, end points of mitochondrial function were determined in unstable (CS-9 and LS-12) and stable (114) hamster fibroblast cell lines derived from GM10115 cells exposed to 10 Gy X rays. Cell lines isolated after irradiation demonstrated a 20-40% loss of mitochondrial membrane potential and an increase in mitochondrial content compared to the parental cell line GM10115. Surprisingly, no differences were observed in steady-state levels of ATP (P > 0.05). Unstable clones demonstrated increased oxygen consumption (two- to threefold; CS-9) and/or increased mitochondrial electron transport chain (ETC) complex II activity (twofold; LS-12). Using Western blot analysis and Blue Native gel electrophoresis, a significant increase in complex II subunit B protein levels was observed in LS-12 cells. Furthermore, immunoprecipitation assays revealed evidence of abnormal complex II assembly in LS-12 cells. Treatment of LS-12 cells with an inhibitor of ETC complex II (thenoyltrifluoroacetone) resulted in significant decreases in the steady-state levels of H(2)O(2) and a 50% reduction in mutation frequency as well as a 16% reduction in CAD gene amplification frequency. These data show that radiation-induced genomic instability was accompanied by evidence of mitochondrial dysfunction leading to increased steady-state levels of H(2)O(2) that contributed to increased mutation frequency and gene amplification. These results support the hypothesis that mitochondrial dysfunction originating from complex II can contribute to radiation-induced genomic instability by increasing steady-state levels of reactive oxygen species.

Mitochondrial electron transport chain blockers enhance 2-deoxy-D-glucose induced oxidative stress and cell killing in human colon carcinoma cells

Increasing evidence suggests that cancer cells (relative to normal cells) have altered mitochondrialelectron transport chains (ETC) that are more likely to form reactive oxygen species (ROS; i.e. O2•-and H2O2) resulting in a condition of chronic metabolic oxidative stress, that maybe compensated forby increasing glucose and hydroperoxide metabolism. In the current study, the ability of an inhibitor ofglucose metabolism, 2-deoxy-D-glucose (2DG), combined with mitochondrial electron transport chainblockers (ETCBs) to enhance oxidative stress and cytotoxicity was determined in human coloncancer cells. Treatment of HT29 and HCT116 cancer cells with Antimycin A (Ant A) or rotenone (Rot)increased carboxy-dichlorodihydrofluorescein diacetate (H2DCFDA) and dihydroethidine (DHE)oxidation, caused the accumulation of glutathione disulfide and enhanced 2DG-induced cell killing. Incontrast, Rot did not enhance the toxicity of 2DG in normal human fibroblasts supporting thehypotheses that cancer cells are more susceptible to inhibition of glucose metabolism in the presenceof ETCBs. In addition, 2-methoxy-antimycin A (Meth A; an analog of Ant A that does not have ETCBactivity) did not enhance 2DG-induced DHE oxidation or cytotoxicity in cancer cells. Finally, in HT29tumor bearing mice treated with the combination of 2DG (500 mg/kg) + Rot (2 mg/kg) the averagerate of tumor growth was significantly slower when compared to control or either drug alone. Theseresults show that 2DG-induced cytotoxicity and oxidative stress can be significantly enhanced byETCBs in human colon cancer cells both in vitro and in vivo.

Nonesterified fatty acid exposure activates protective and mitogenic pathways in vascular smooth muscle cells by alternate signaling pathways

Vascular smooth muscle cells (VSMC) are dynamic cells exposed to fluctuating concentrations of nutrients on a daily basis. Nonesterified fatty acids (NEFA) have been indicted as potential mediators of atherosclerosis and exaggerated VSMC remodeling observed in diabetes, and in vitro data support a model of VSMC activation by NEFA. However, recent observations suggest that metabolic stressors such as oxidants and NEFA may also simultaneously induce cytoprotective events as part of a homeostatic “off switch.” Our group has established that the transcription factor cyclic adenosine monophosphate response element binding protein (CREB) is important for maintenance of VSMC quiescence, differentiation, and survival. We therefore examined whether acute physiologic NEFA exposure would regulate CREB in primary cultures of bovine aortic VSMC and explored the relationship between signaling to the cytoprotective CREB and the activating mitogen-activated protein kinase pathways. In vitro exposure of VSMC to 3 classes of unsaturated NEFA leads to significant acute, transient, dose-dependent, and repeatedly inducible CREB activation. As expected, extracellular signal–regulated kinase, P38 mitogen-activated protein kinase, Akt, Jun N-terminal kinase, and protein kinase C (PKC) pathways are also activated by NEFA. Using a battery of pharmacologic inhibitors and antioxidants, we demonstrate that CREB activation is mediated by a novel PKC isoform and is reactive oxygen species independent, whereas extracellular signal–regulated kinase activation, in contrast, is mediated by reactive oxygen species and is PKC independent. These data suggest parallel and mechanistically distinct stimulation of separate stabilizing and activating pathways in VSMC response to acute NEFA-mediated stress. Furthermore, the down-regulation of CREB in models of chronic metabolic stress reported in the literature would be expected to disrupt this homeostasis and shift the balance toward VSMC activation, consistent with emerging models of atherosclerosis.

The rationale for cardiomyocyte resuscitation in myocardial salvage

Clinical heart failure results from the cumulative loss of functioning myocardium from any cause. At the cellular level, cardiac myocytes die from three causes, individually or in combination: Necrosis occurs when external conditions are not sufficient to sustain minimal cellular functions, as with ischemia, and there is a general and unorganized breakdown of cell organelles, engendering an inflammatory response that may have harmful collateral tissue effects. Apoptosis, or cell suicide, occurs when specific external or internal conditions provoke a highly structured sequence of events to shut down cellular functions and remove the cell, with minimal consequences to surrounding tissue. Autophagy is a normal response to cell starvation that is induced under conditions of chronic metabolic or other stress. Current therapeutics, such as early myocardial revascularization after myocardial infarction, are focused exclusively upon minimizing cardiac myocyte necrosis and may even contribute to secondary apoptosis and autophagy. This review explores possible approaches to bring cardiac myocytes that are destined to die, back to life, i.e., cellular resuscitation. Two pro-apoptotic proteins in particular, Bnip3 and Nix, are transcriptionally upregulated specifically in response to myocardial ischemia and pathological hypertrophy and have been examined as therapeutic targets. In Bnip3 and Nix genetic mouse models, prevention of cardiac myocyte apoptosis in ischemic and hemodynamically overloaded hearts salvaged myocardium, minimized late ventricular remodeling, and enhanced ventricular performance. Cardiomyocyte resuscitation by preventing programmed cell death shows promise as an additive approach to minimizing necrosis for long-term prevention of heart failure.

The transcription of MGAT4A glycosyl transferase is increased in white cells of peripheral blood of Type 2 Diabetes patients

BackgroundHuman glycosylase IV is involved in GLUT2 transporter regulation in pancreatic β cells. A KO of this gene along with a high fat diet in a mice model has been associated with the development of type 2 diabetes (T2D). The aims of this study were to measure and compare the MGAT4A mRNA levels in white blood cells (WBC) from T2D subjects and healthy subjects (T2NB), and to measure the half-life of the MGAT4A mRNA.ResultsWe studied a sample of 73 individuals, 40 T2D subjects and 33 T2NB subjects. Anthropometrical and biochemical profiles were registered. The MGAT4A mRNA levels in WBC and the transcript half-life in Jurkat T cells were determined by Real-Time PCR. A blood differential cell counting was made for each individual. Cell counting showed T2D subjects exhibited an increased number of WBC compared to T2NB subjects (P = 0.0001). Biochemical parameters such as fasting glucose (P = 0.0001), and triglycerides (P = 0.002) were statistically significant. T2D subjects had 4.2-fold more MGAT4A transcript compared to T2NB subjects (P = 0.002). The MGAT4A mRNA had a half-life of 2.04 h in Jurkat T cells.ConclusionThe results of this work suggest that in T2D subjects, high levels of glucose and triglycerides are accompanied by an increase on MGAT4A mRNA levels and WBC count; condition that suggests a pro-inflammatory state due to a chronic metabolic stress.

The phosphatidylethanolamine N-methyltransferase pathway is quantitatively not essential for biliary phosphatidylcholine secretion

The phosphatidylethanolamine N-methyltransferase (PEMT) pathway of phosphatidylcholine (PC) biosynthesis is not essential for the highly specific acyl chain composition of biliary PC. We evaluated whether the PEMT pathway is quantitatively important for biliary PC secretion in mice under various experimental conditions. Biliary bile salt and PC secretion were determined in mice in which the gene encoding PEMT was inactivated (Pemt(-/-)) and in wild-type mice under basal conditions, during acute metabolic stress (intravenous infusion of the bile salt tauroursodeoxycholate), and during chronic metabolic stress (feeding a taurocholate-containing diet for 1 week). The activity of CTP:phosphocholine cytidylyltransferase, the rate-limiting enzyme of PC biosynthesis via the CDP-choline pathway, and the abundance of multi-drug-resistant protein 2 (Mdr2; encoded by the Abcb4 gene), the canalicular membrane flippase essential for biliary PC secretion, were determined. Under basal conditions, Pemt(-/-) and wild-type mice exhibited similar biliary secretion rates of bile salt and PC ( approximately 145 and approximately 28 nmol/min/100 g body weight, respectively). During acute or chronic bile salt administration, the biliary PC secretion rates increased similarly in Pemt(-/-) and control mice. Mdr2 mRNA and protein abundance did not differ between Pemt(-/-) and wild-type mice. The cytidylyltransferase activity in hepatic lysates was increased by 20% in Pemt(-/-) mice fed the basal (bile salt-free) diet (P < 0.05). We conclude that the biosynthesis of PC via the PEMT pathway is not quantitatively essential for biliary PC secretion under acute or chronic bile salt administration.

Methyl deficiency, alterations in global histone modifications, and carcinogenesis.

The methyl-deficient model of endogenous hepatocarcinogenesis in rodents is unique in that dietary omission rather than the addition of chemical carcinogens leads to tumor formation. Thus, the biochemical and molecular events predisposing to cancer in this model result from chronic metabolic stress and provide an ideal model system to study progressive alterations that occur during carcinogenesis. Moreover, epigenetic alterations imposed by this diet are believed to be 1 of the main mechanisms responsible for malignant transformation of rat liver cells. In this study we examined the changes in global histone modification patterns in liver during hepatocarcinogenesis induced by methyl deficiency. Feeding animals the methyl-deficient diet (MDD) led to progressive loss of histone H4 lysine 20 trimethylation (H4K20me3), H3 lysine 9 trimethylation (H3K9me3), and histone H3 lysine 9 (H3K9ac) and histone H4 lysine 16 (H4K16ac) acetylation. A considerable decrease of H4K20me3 and H3K9ac was also detected in liver tumors induced by MDD. In contrast, liver tumors displayed an increase in H3K9me3 and H4K16ac. To determine the possible mechanism of alterations of histone modifications, we analyzed the expression of histone-modifying enzymes in liver during hepatocarcinogenesis. The expression of Suv4-20h2 and RIZ1 histone methyltransferases (HMTs) steadily decreased along with the development of liver tumors and reached its lowest level in tumor tissue, whereas the expression of Suv39-h1 HMT and histone acetyltransferase 1 (HAT1) substantially increased in tumors. These results illustrate the complexity and importance of histone modification changes in the etiology of hepatocarcinogenesis induced by MDD.

Corticotropin Releasing Hormone: a Diagnostic Marker for Behavioral and Reproductive Disorders?

Corticotropin-releasing hormone (CRH) is a key mediator of endocrine, autonomic, behavioral, and immune responses to stress. The ability of CRH to induce hormonal stress responses has been used to investigate the functionality of the hypothalamus-pituitary-adrenal axis, and consequently the activity of hypothalamic CRH neuronal systems. Indeed, CRH administration to humans causes prompt release of ACTH, followed by secretion of cortisol, aldosterone and other adrenal steroids. CRH hypersecretion/hyperactivity has been associated to major depression, anxiety-related disorders, anorexia nervosa, Alzheimer's and Parkinson's diseases and progressive supranuclear palsy. During pregnancy the human placenta and its accessory membranes are the major sites of CRH synthesis and secretion. Placental CRH secretion is autonomous, but increasing evidence indicates that maternal or fetal conditions may influence such secretion. Therefore, the emerging concept is that in the event of acute or chronic metabolic, physical or infectious stress, the placenta takes part in a stress syndrome by releasing CRH, which may contribute to restore local blood flow and to influence the timing of delivery. The CRH released by the placenta is measurable in maternal plasma and other biological fluids and may be used to diagnose subclinical processes anteceding pregnancy complications such as pre-eclampsia and preterm delivery.

Bilateral Central Crystalline Corneal Deposits Four Years After Intacs for Myopia

To report a case of bilateral central crystalline keratopathy in the anterior stroma occurring 4 years after Intacs implantation. A 45-year-old woman underwent bilateral uncomplicated Intacs implantation for myopia. The postoperative course was uneventful. However, between 3 and 4 years after surgery, the patient developed central opacifications of the anterior stroma in both eyes, reducing best spectacle-corrected visual acuity. Intacs were explanted. Confocal microscopy, electron microscopy of the explanted ring segments, and microbiology studies were performed. Opacities were still detectable at the slit-lamp microscope up to 8 months after explantation. This is the first report on central corneal opacifications after Intacs implantation for myopia. The opacities could be the result of chronic metabolic stress or the beginning of lipid-like changes in another more central corneal localization.