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Molecular cloning of the heat-shock cognate 70 (Hsc70) gene from the two-spotted spider mite, Tetranychus urticae, and its expression in response to heat shock and starvation.

We isolated a heat shock cognate 70 (hsc70) gene from the two-spotted spider mite, Tetranychus urticae, a serious agricultural pest. The hsc70 cDNA is 2275 bp and contains a 1962 bp open reading frame. The translated amino acid sequence consists of 654 residues with a calculated molecular mass of 71,275 Da and an isoelectronic point (pI) of 5.52. It also contains the highly conserved functional motifs of the Hsp70 family. A comparison of the deduced amino acid sequence shows a high identity (81-84%) with Hsp70s/Hsc70s of insects but the highest identity is with mussel Hsc71 (86%). Northern blot hybridization indicates that the hsc70 transcript level of female adults is higher than that of male adults. We evaluated the response of hsc70 gene to stresses from temperature and starvation. The level of hsc70 mRNA was not significantly changed by heat and cold shocks nor by recovery after the shocks. However, the hsc70 mRNA level was decreased by food restriction of female mites. Analysis of nucleotide and deduced amino acid sequences of hsc70 gene from T. urticae suggests that it is a member of heat shock cognate 70 gene in the highly conserved Hsp70 family but that its expression is influenced by food restriction rather than thermal stress. This is the first molecular analysis of a heat shock protein gene in an acarid.

Amino Acid Sequence↗

Mapping out starvation responses.

What are the pathways that underlie the coordinated responses of an organism to well-fed and food-deprived states? A report in this issue of Cell Metabolism suggests that starvation functions via a muscarinic acetylcholine receptor to activate MAP kinase signaling in the pharyngeal muscle of C. elegans (You et al., 2006).

Acetylcholine↗

Feeding glucose or sucrose, but not trehalose, suppresses the starvation-induced premature pupation in the yellow-spotted longicorn beetle, Psacothea hilaris.

Under 25 degrees C and a long-day photoperiod, starvation induces premature pupation in 4th instar Psacothea hilaris larvae exceeding a threshold weight of 180 mg, resulting in the formation of small but morphologically normal adults. To investigate possible mechanisms underlying this phenomenon, we first measured the hemolymph trehalose and glucose levels of starved larvae. When larvae were starved after 4 days of feeding (attaining the threshold weight), glucose levels decreased 4-fold within the next 24 h, while trehalose levels, after a temporary slight decrease, increased remarkably to reach a peak just before the prepupa stage. The effects of ingesting various nutrients on the developmental fate and the hemolymph sugar titers of starving larvae were then examined. Feeding on agar blocks containing sucrose or glucose totally suppressed the occurrence of premature pupation, while trehalose, fructose, casein and starch were ineffective. Feeding on glucose or trehalose resulted in a 6-fold decrease in hemolymph glucose levels and remarkably elevated trehalose levels. Since feeding on glucose and trehalose induced similar changes in hemolymph sugar titers but trehalose was not effective in suppressing premature pupation, glucose may have exhibited its effects via gustatory mechanisms.

Animal Nutritional Physiological Phenomena↗

Starvation period and age affect the response of female Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) to odor and visual cues.

The effects of starvation or age on the walking or flying response of female Frankliniella occidentalis to visual and/or odor cues in two types of olfactometer were examined in the laboratory. The response of walking thrips starved for 0, 1, 4, or 24h to an odor cue (1microl of 10% p-anisaldehyde) was examined in a Y-tube olfactometer. The take-off and landing response of thrips (unknown age) starved for 0, 1, 4, 24, 48 or 72h, or of thrips of different ages (2-3 days or 10-13 days post-adult emergence) starved for 24h, to a visual cue (98 cm(2) yellow sticky trap) and/or an odor cue (0.5 or 1.0 ml p-anisaldehyde) was examined in a wind tunnel. More thrips walked up the odor-laden arm in the Y-tube when starved for at least 4h (76%) than satiated thrips (58.7%) or those starved for 1h (62.7%, P<0.05). In the wind tunnel experiments the percentage of thrips to fly or land on the sticky trap increased between satiated thrips (7.3% to fly, 3.3% on trap) and those starved for 4h (81.2% to fly, 29% on trap) and decreased between thrips starved for 48 (74.5% to fly, 23% on trap) and 72 h (56.5% to fly, 15.5% on trap, P<0.05). Fewer younger thrips (38.8%) landed on a sticky trap containing a yellow visual cue of, those that flew, than older thrips (70.4%, P<0.05), although a similar percentage of thrips flew regardless of age or type of cue present in the wind tunnel (average 44%, P>0.05).

Age Factors↗

Starvation in humans: evolutionary background and contemporary implications.

Although there is extensive evidence that caloric restriction (CR) extends lifespan in several species the evidence base for humans is weak. We are still at the stage of applying inductive reasoning and of framing hypotheses to be tested. It is known that a genetic background contributes about 25% to the variation in human longevity, but thought unlikely that any genes conferring longer lifespan have been positively selected to do so. It is more likely that any such benefits are unintended consequences arising from other adaptations. If there is an association between CR and longevity in humans it may have been selected by previous exposures to famine. This paper briefly reviews the historical evidence on the extent and frequency of famines in human history. It is concluded that starvation has been one of the major selective pressures on the human genome and has left abundant evidence of adaptive survival traits. Many of these are mediated through effects on reproduction. However, interpretation of the possible links between these energy-sparing mechanisms and any association between CR and ageing is handicapped by an absence of data on the latter and will remain a matter of debate for many years to come.

Aging↗

Reduced leptin levels in starvation increase susceptibility to endotoxic shock.

Malnutrition compromises immune function, reducing resistance to infection. We examine whether the decrease in leptin induced by starvation increases susceptibility to lipopolysaccharide (LPS)- and tumor necrosis factor (TNF)-induced lethality. In mice, fasting for 48 hours enhances sensitivity to LPS. Decreasing the fasting-induced fall in leptin by leptin administration markedly reduced sensitivity to LPS. Although fasting decreases basal leptin levels, LPS treatment increased leptin to the same extent as in fed animals. Fasting increased basal serum corticosterone; leptin treatment blunted this increase. Fasting decreased the ability of LPS to increase corticosterone; leptin restored the corticosterone response to LPS. Serum glucose levels were decreased in fasted mice and LPS induced a further decrease. Leptin treatment affected neither basal glucose nor that after LPS. LPS induced a fivefold greater increase in serum TNF in fasted mice, which was blunted by leptin replacement. In contrast, LPS induced lower levels of interferon-gamma and no differences in interleukin-1beta in fasted compared to fed animals; leptin had no effect on those cytokines. Furthermore, fasting increased sensitivity to the lethal effect of TNF itself, which was also reversed by leptin treatment. Thus, leptin seems to be protective by both inhibiting TNF induction by LPS and by reducing TNF toxicity.

Animals↗

Starvation-induced posttranscriptional control of rat liver mitochondrial citrate carrier expression.

Starvation has been associated with a reduced citrate carrier (CTP) activity in rat liver mitochondria. In the present study the molecular mechanism responsible for this reduction was investigated. Northern blot analysis performed with hepatic total RNA showed a decrease of about 40% in the CTP mRNA abundance in starved rats, when compared to fed animals. Nuclear run-on assay did not reveal any appreciable difference in the rate of CTP mRNA synthesis between the two groups of animals, while the apparent half-life of CTP mRNA in hepatocytes from fed and starved rats was 11 and 6h, respectively. Therefore, these results suggest that in starved rats the regulation of hepatic CTP expression occurs at posttranscriptional level. Moreover, the reduced CTP activity in starved animals gradually increased by refeeding. The carrier activity reached fed rat values 6-9h following refeeding. Interestingly, the accumulation of CTP mRNA raised in parallel with the transport activity.

Animals↗

Effect of starvation and chlormethiazole on cytochrome P450s of rat nasal mucosa.

Cytochrome P450 (CYP) enzymes of nasal tissue are relatively resistant to induction by classical inducers. In the present study, the effects of starvation on the expression of CYP1A, 2A, 2B, 2C, 2E, 2G, and 3A subfamilies in the nasal mucosa of rat were studied. Fasting for 72 hr caused an increase in 2E1-dependent p-nitrophenol hydroxylase and 1A-dependent ethoxy- (or methoxy) resorufin dealkylase activities, but did not affect either 2A-linked coumarin hydroxylase or the testosterone hydroxylase activity, the latter reaction being a marker of several CYPs including 2G1. Whereas increases in 2E1- and 1A- associated catalytic activities were accompanied by a concomitant increase in the corresponding apoproteins as determined by immunoblotting, immunoactive protein bands reactive with antibodies raised against rat 1A1, 2B1, 2C11, 3A1 or rabbit nasal 2A10/11 and 2G1 were not altered. Fasting also increased CYP2E1 and CYP1A2 on the mRNA level, but did not alter CYP1A1 mRNA as determined by hybridization with cDNA probes selective for these cytochromes. A reiterative administration of chlormethiazole, a specific inhibitor of 2E1 in liver, strongly inhibited many CYPs, including 2E1, 1A2, 2G1, and 2A in the nasal mucosa, but did not influence expression of 2B or 3A as determined by immunoblotting or catalytic activities. The chlormethiazole-mediated inhibition of 1A1 and 2E1 was demonstrated to be at the mRNA level. These results suggest that fasting induces the gene expression of 2E1 and 1A2 and that the mechanisms involved in the regulation of CYPs in the nasal mucosa are tissue-specific. The inducibility of the above-mentioned isoforms may have a significant role in the clearance of drugs and bioactivation of inhaled compounds.

Animals↗

Molecular cloning and cellular distribution of two 14-3-3 isoforms from Hydra: 14-3-3 proteins respond to starvation and bind to phosphorylated targets.

In the simple metazoan Hydra a clear link between food supply and cell survival has been established. Whilst in plants 14-3-3 proteins are found to be involved in signalling cascades that regulate metabolism, in animals they have been shown to participate in cell survival pathways. In order to explore the possibility that 14-3-3 proteins in Hydra could be involved in regulating metabolism under different conditions of food supply, we have cloned two isoforms of 14-3-3 proteins. We show here that 14-3-3 proteins bind to phosphorylated targets in Hydra and form homo- and heterodimers in vitro. 14-3-3 proteins are localised in the cytoplasm of all cells and also in the nuclei of some epithelial cells. This nuclear localisation becomes more prominent during starvation. Moreover, 14-3-3 protein is present in large amounts in food granules and from this we conclude that it performs functions which are associated with metabolism and food storage in Hydra.

14-3-3 Proteins↗

Stereological study on the rat heart in chronic alimentary thiamine deficiency--absence of myocardial changes despite starvation.

Young male Wistar rats were fed with a thiamine-free diet for 35 days. The growth of the thiamine-deficient animals was considerably suppressed, as compared to a pair-fed control group (body weights: 140.4 +/- 4.3 g v. 243.1 +/- 4.3 g). A weight-matched control group was therefore evaluated in addition. Histological and ultrastructural investigations of the hearts did not reveal any morphological abnormalities. Stereological analysis was performed on left ventricular papillary muscles. One-way analysis of variance showed significant differences of stereological parameters between the three groups which were not associated with significant linear contrasts between the thiamine-deficient group and the weight-matched control group. Thus, the changes are not to be related to direct effects of thiamine depletion on the myocardium. Conclusively, the data indicate that stereological parameters (ratios) of the myocardium depend on heart weight and body weight, respectively, and are changed during normal growth. Our stereological estimates disclosed alterations of the following parameters: numerical density of myocardial cell nuclei, length density and volume density of capillaries, surface density of outer mitochondrial membranes and surface-to-volume ratio of mitochondria, and volume density of myofibrils. Furthermore, relative heart weights decrease in normal growth. It is concluded that our experimental model of chronic alimentary thiamine deficiency was associated with a normal structure of the myocardium. All significant differences between the thiamine-deficient group and the control group are explained by body weight changes following starvation in chronic thiamine deficiency.

Animals↗

Extracellular ATP inhibits starvation-induced apoptosis via P2X2 receptors in differentiated rat pheochromocytoma PC12 cells.

Apoptosis in neuronal tissue is an efficient mechanism which contributes to both normal cell development and pathological cell death. The present study explored the effects of extracellular ATP on starvation-induced apoptosis in rat pheochromocytoma PC12 cells. Incubation of differentiated PC12 cells with ATP for 6h suppressed apoptosis. 2-Methylthio-ATP, a P2 purinoceptor agonist, was as potent as ATP in suppressing apoptosis, whereas adenosine, ADP, alpha,betamethylene-ATP or UTP was totally ineffective. The suppressive action of ATP was dependent upon the presence of extracellular Ca2+ and blocked by co-incubation with the P2 antagonist, suramin. DNA ladder formation, a typical symptom of apoptosis in starved cells, was inhibited by ATP, 2-methylthio-ATP but not by UTP. These results suggest that the inhibitory action of extracellular ATP on apoptotic cell death is mediated via the activation of P2X2 receptors in differentiated PC12 cells.

Adenosine Triphosphate↗

Nutritional consequences of major gastrointestinal surgery. Patient outcome and starvation.

Advances in major gastrointestinal surgery and the existence of a significant number of high-risk patients necessitate a detailed knowledge of the metabolic response to surgery and the need for adjunctive nutritional therapy. Such surgery has the potential to cause significant nutritional consequences affecting patient outcome. Often, patients present malnourished preoperatively because of their chronic gastrointestinal disease, advanced age, or comorbidity. Still others are at great risk for developing malnutrition postoperatively secondary to the hypercatabolism of prolonged illness or postoperative complications. In addition, the surgical alterations of the gastrointestinal tract might disrupt normal function sufficiently to result in chronic nutritional deficiencies. Because starvation is no longer an acceptable condition, these concerns need to be recognized prior to surgery, and if indicated, nutritional support must be initiated preoperatively. Placement of feeding jejunostomy catheters for early enteral support should be considered for all high-risk patients. Equally important, surgeons should maintain a watch for future nutritional innovations that will improve their ability to meet patients' nutritional requirements.

Anorexia↗

Starvation suppression and refeeding activation of infection. An ecological necessity?

The hypothesis is advanced that starvation suppresses and refeeding activates certain infections as an essential part of an ecological balance between man, his animals, and his environment. During famine, then, man fails to thrive, but his ultimate extinction is prevented in part by the parallel decline in fecundity of his "micropredators". In times of plenty the parallel increase in the same predators is a check against his excessive multiplication.

Adult↗

"Accelerated starvation" and the skipped breakfast in late normal pregnancy.

After dinner at 6 P.M. food was withheld from non-pregnant women and women in the third trimester of pregnancy: there were lean and obese women in each group, and all had normal carbohydrate metabolism. Levels of circulating fuels and glucoregulatory hormones were monitored between 6 A.M. (12 h fast) and noon (18 h fast) the following day. In non-pregnant women plasma levels of glucose, alanine, free fatty acids (FFA) beta-hydroxybutyrate, insulin, and glucagon remained constant between 6 A.M. and noon except for slight fall in plasma alanine in the obese women and slight increases in beta-hydroxybutyrate in the lean. By contrast, in pregnant women the concentrations of all substances measured changed profoundly after extension of the overnight fast. Levels of plasma glucose, which were significantly lower than those in non-pregnant women at 6 A.M., fell further during the subsequent 6 h; plasma alanine levels declined significantly during the same interval, and there were coincident rises in plasma FFA and beta-hydroxybutyrate and reductions in plasma insulin. The changes in lean and obese women were of similar magnitude; habitus did not therefore seem to have effects additional to those of pregnancy on the response to skipping breakfast. In all the pregnant women final levels of FFA and beta-hydroxybutyrate were strongly correlated and inversely related to final levels of plasma glucose. The studies indicate that "accelerated starvation" can be unmasked during pregnancy even with the minor dietary deprivation common in conventional clinical circumstances. Thus, insofar as it may be desirable to avoid ketonaemia during pregnancy, the common practice of skipping breakfast whether by personal preference or for doctor-initiated laboratory testing should be avoided in pregnant women.

3-Hydroxybutyric Acid↗

Starvation-induced hyperactivity in the rat: the role of endocrine and neurotransmitter changes.

Semistarved rats develop high running wheel activity. This running activity induces increased norepinephrine, dopamine, and serotonin turnover in the hypothalamus. Corticosterone in plasma becomes increased while luteinizing hormone and testosterone are suppressed. In female rats cyclic gonadal function is suppressed. Running activity in the semistarved rats can be suppressed specifically by serotonin 1-c-agonists and by alpha 2-adrenoceptor agonists. This animal model is helpful in the understanding of the combined effects of starvation and hyperactivity, which are observed in many patients with anorexia nervosa. The observation of the serotonergic system might help to develop a pharmacological treatment of hyperactivity in anorectic patients.

Animals↗

Advancing monosaccharides as biomarkers: Part II. Effects of starvation and cadmium in Chironomus riparius as detected by fluorophore-assisted carbohydrate-electrophoresis.

Saccharides were evaluated as biomarkers for cadmium (Cd) and starvation using fluorophore-assisted carbohydrate-electrophoresis (FACE) in 4th instar Chironomus riparius. FACE allowed different types of saccharides in whole larval homogenate to be analyzed simultaneously and in parallel with other larval samples. Larval homogenates showed seven principle bands labeled A, B, C, D, E, F and G. Previous work found that the migration patterns of bands A, C, D and F matched those of ribose, glucose, galactose and fructose, respectively. Four of the bands, B, C, E and G were generated from glucose-based mono, oligo, and polysaccharides. Band B was primarily derived from glucose and band E from glycogen. Experiments (0-72 h) with starved larvae showed a time dependent reduction in bands B and E that was statistically significant at 72 h. Experiments with Cd (0.01-1000 microM) showed a concentration and time dependent reduction in band E with a LOEL of 1 microM and NOEL of 0.01 microM at 48 h. The LOEL was 0.014% of the 48 h LC50. Significant reduction of band E only occurred in fed larvae indicating that food was an important route of exposure. Reductions in saccharides were independent of larval weight loss at 48 h. This suggested that major changes in saccharides were not due to weight loss but metabolic stress in the presence of Cd.

Animals↗

Tolerance and starvation induced cross-protection against different stresses in Aeromonas hydrophila.

Aeromonas hydrophila is sometimes considered as a controversial human pathogen and reported to be susceptible to food processing procedures and environmental stresses. In this study, we have shown that early stationary phase cells of A. hydrophila were readily killed during up shifts in temperature (in the range 50-70 degrees C), the course of drying (at relative humidity, temperature and brightness of the laboratory) and after 5 min exposure to 20%, 30% and 40% v/v ethanol. However, this bacterium was found moderately susceptible to down shift to 4 degrees C in nutrient poor water, sodium chloride stresses (1.5 and 2 M) and to 12% and 15% v/v ethanol stresses. Tolerance against 1 M NaCl and 10% v/v ethanol was observed. At ambient temperature (24.5 degrees C), this microorganism exhibited a starvation survival state, which was largely independent of the initial cell concentrations (8.82, 7.71 and 6.76 log units). The cross-protection experiments showed that cells starved for short (1 day) or prolonged (50 days) periods developed increased resistance to down shift at 4 degrees C and ethanol stress. This may be of concern to the food-processing industry from the public health perspective.

Adaptation, Physiological↗

Presence of morphine and morphine-6-glucuronide in the marine mollusk Mytilus edulis ganglia determined by GC/MS and Q-TOF-MS. Starvation increases opiate alkaloid levels.

Morphine and morphine-6-glucuronide, a morphine metabolite, have been identified and quantified in Mytilus edulis pedal ganglia at a level of 2.67+/-0.44 and 0.98+/-0.14 ng/ganglia, respectively by high performance liquid chromatography coupled to electrochemical detection. These opiate alkaloids were further identified by both gas-chromatography mass spectrometry and nanoflow electrospray ionization double quadrupole orthogonal acceleration Time of Flight mass spectrometry. In animals that were starved, the morphine level rose to 6.38+/-0.88 ng/ganglion and the morphine 6-glucoronide rose to a level of 23.0+/-3.2 ng/ganglion after 30 days. These studies demonstrate that opiate alkaloids are present as naturally occurring signal molecules whose levels respond to stress, i.e., starvation. Opiate alkaloids were not found in the animal's incubation media or food, demonstrating their synthesis occurred in the respective tissue. These new method of opiate alkaloid detection, conclusively proves that morphine and morphine-6-glucuronide are present in animal tissues.

Animals↗