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Non-Mammalian Vertebrates: Distinct Models to Assess the Role of Ion Gradients in Energy Expenditure
- School of Animal and Comparative Biomedical Science, University of Arizona, Tucson, AZ, United States
Animals store metabolic energy as electrochemical gradients. At least 50% of mammalian energy is expended to maintain electrochemical gradients across the inner mitochondrial membrane (H+), the sarcoplasmic reticulum (Ca++), and the plasma membrane (Na+/K+). The potential energy of these gradients can be used to perform work (e.g., transport molecules, stimulate contraction, and release hormones) or can be released as heat. Because ectothermic species adapt their body temperature to the environment, they are not constrained by energetic demands that are required to maintain a constant body temperature. In fact, ectothermic species expend seven to eight times less energy than similarly sized homeotherms. Accordingly, ectotherms adopt low metabolic rates to survive cold, hypoxia, and extreme bouts of fasting that would result in energy wasting, lactic acidosis and apoptosis, or starvation in homeotherms, respectively. Ectotherms have also evolved unique applications of ion gradients to allow for localized endothermy. Endothermic avian species, which lack brown adipose tissue, have been integral in assessing the role of H+ and Ca++ cycling in skeletal muscle thermogenesis. Accordingly, the diversity of non-mammalian vertebrate species allows them to serve as unique models to better understand the role of ion gradients in heat production, metabolic flux, and adaptation to stressors, including obesity, starvation, cold, and hypoxia.
The ease of genetic manipulation, the standard husbandry demands, and the well-established experimental methods have resulted in rodent model predominating the obesity field. Yet, the diversity of non-mammalian vertebrates, including both endothermic (birds and some fish) and ectothermic (amphibians, reptiles, and fish) species that have adapted to a variety of environments, provides unique opportunities to better understand the regulation of energy expenditure. The heterogeneity of species and environments to which they have adapted yield an abundance of unique attributes that can better help us understand energy expenditure. Herein, we review research in non-mammalian vertebrate control of electrochemical gradients and application of this work to mammalian species.
Basal metabolic rate is seven- to eightfold higher in mammals than in similarly sized ectotherms maintained at 37°C (1, 2). Because ectothermic species have low levels of basal heat production, they are sensitive models to identify the role of ion flux in altering heat production and basal metabolic rate. For example, egg brooding female pythons use whole body skeletal muscle contractions to facilitate endothermy and maintain a body temperature 9–13°C above ambient temperature (3, 4). This contraction-induced thermogenesis increases the oxygen consumption 22 times above baseline (5). In rat pups (22 days), maintenance of body temperature in an environment 9–13°C, colder than thermoneutral, increases oxygen consumption just two times above baseline (6). While in penguin chicks, a temperature drop of 10°C below the shivering threshold only increases oxygen consumption by 27%, while a temperature 25°C below the shivering threshold doubles oxygen consumption (7). Thus, the low basal metabolic rate of ectothermic vertebrate species allow for a uniquely sensitive model to perturbations in metabolic rate, while the endothermic bird provides a comparative model to better understand perturbations and adaptations that affect whole body and skeletal muscle energy expenditure in the absence of endothermic brown adipose tissue.
Herein, we review the lower vertebrate literature on energy expenditure with a focus on three ion gradients: the H+ gradient at the inner mitochondrial membrane, the Ca++ gradient of the sarcoplasmic reticulum, and the Na+/K+ gradient across the plasma membrane. We further apply findings from the lower vertebrate literature to our current understanding of mammalian energy expenditure and its potential application to human health.
The H+ gradient is maintained in the inter-mitochondrial membrane space, created by electron transport chain activity (Figure 1). This gradient is dissipated by H+ ion leak across the inner mitochondrial membrane, which can be exacerbated by uncoupling proteins (UCP) and adenine nucleotide translocases (ANT), or production of adenosine triphosphate (ATP) through ATP synthase. Accordingly, the mitochondrial density, inner-mitochondrial membrane surface area, expression and activity of electron transport chain proteins, and the expression of UCPs and ANTs that allow for H+ leak across the inner mitochondrial membrane are integral to heat production and energy expenditure.
Figure 1. Schematic representation of the Ca++ gradient at the sarcoplasmic reticulum, H+ gradient at the inner mitochondrial membrane, and Na+/K+ gradient at the plasma membrane. Flames indicate sites of heat generation from ion leak and non-functional adenosine triphosphate (ATP) hydrolysis. Abbreviations: SERCA, sarco/endoplasmic reticulum Ca++ ATPase; RYR, ryanodine receptor; SLN, sarcolipin; ETC, electron transport chain; UCP, uncoupling protein; ANT, adenine nucleotide translocase.
Compared to endotherms, reptilian tissues have fewer mitochondria with less inner mitochondrial membrane surface area per mitochondria, resulting in 50% less inner mitochondrial membrane across which H+ can leak (1). Because the organs that are rich in mitochondria are smaller in ectotherms, whole body mitochondrial membrane surface area is four times greater in mammals than in reptiles of the same body size (8). The H+ gradient is established by electron transport chain activity and depressed by the release of H+ out of the inter-mitochondrial membrane space. With four times less membrane surface area, ectotherms have lower electron transport chain activity and H+ ion leak.
Electron Transport Chain
The decreased electron transport chain activity in ectotherms is not a result of decreased activity of proteins integral to oxidative phosphorylation (2, 9). ATP synthesis, expressed as a percentage of mitochondrial respiration, is similar in reptiles and mammals at similar body temperature (2). Inner mitochondrial membrane cytochrome C oxidase (complex IV) content and activity is also similar in fish and cattle (9). Moreover, when corrected for tissue protein content, cytochrome C oxidase activity is similar in reptiles and mammals (10). Thus, the decreased electron transport chain activity in ectotherms is a result of decreased mitochondria numbers, mitochondrial size, mitochondrial membrane area, and a resulting decrease in total electron transport proteins rather than altered activity of the mitochondrial machinery.
The electron transport chain represents an important point of manipulation for energy expenditure in ectotherms. During hypoxia and cold exposure, electron transport chain activity can be depressed to accommodate the decreased cellular energy demand imposed by these conditions. Thus, in species not dependent on internal heat production, limiting cellular respiration is an adaptive response to environmental stress. In fact, ectothermy allows for adaptive whole body cooling and metabolic depression to prevent metabolic acidosis in response to hypoxia (11). Four months of in vivo hypoxia results in a severe decrease in adenosine diphosphate (ADP) stimulated respiration (State 3) and a decrease in ADP-independent respiration (State 4; H+ leak) from isolated skeletal muscle mitochondria (12). Hypoxia induces a characteristic rise in plasma lactate in frogs maintained at 7°C, while there is no change in plasma lactate in frogs maintained at 1.5°C (13). The rise in lactate at 7°C is a result of glycolytic flux exceeding the mitochondrial capacity for oxidation under hypoxic conditions. In turn, frogs or toads challenged with hypoxia chose to reside at lower temperatures, decrease their metabolic rate, and limit the metabolic disturbances associated with a lack of oxygen (13–15). In alligators, hypoxia leads to hypothermia in an adaptive effort to depress metabolic demands (16). Long-term exposure to hypoxia and 3°C water results in an adaptive decrease in skeletal muscle mitochondrial oxygen consumption by inhibiting flux through the electron transport chain and limiting H+ ion leak across the inner mitochondrial membrane (12, 17–19). Indeed, ectothermic species exploit the decrease in metabolic demand that accompanies low body temperatures to adapt to acute or chronic hypoxic environments (14).
Chronic hypoxia, due to a high altitude habitat, also regulates mitochondrial respiration. In a comparison of two closely related lizard species that live at different elevations, species adapted to high elevations had lower oxidative capacity and less ADP-independent respiration than species that inhabit lower elevations (20). Thus, adaptation to chronic hypoxia increased the efficiency of H+ ion gradient energy capture as ATP. Of note, mitochondria from lizards adapted to high elevations are less able to increase respiration in response to an increase in incubation temperature (20). Suggesting that there is an adaptive cost to the tight coupling between the H+ ion gradient and ATP synthesis.
In endotherms, cold exposure increases electron transport chain activity to promote endogenous heat production. Acute, 24 h, cold exposure in the chick increases expression of cytochrome C oxidase and NADH ubiquinone oxidoreductase (complex I) (21). Chronic cold exposure similarly increases cytochrome C oxidase activity as a result of increased mitochondria number and increased inner mitochondrial membrane area/mitochondrial volume (22). In small mammals, the primary thermogenic organ is brown adipose tissue. Chronic, 4 week, cold exposure in the tree shrew increases the cytochrome oxidase activity in brown adipose tissue to six times that observed in thermoneutral conditions (23). The same 4-week cold exposure increases liver cytochrome oxidase activity 2.7 times (23). In mice, skeletal muscle electron transport activity is increased by cold acclimation (24). Mice that lack UCP1 and the thermogenic potential of brown adipose tissue, display an increase in skeletal muscle mitochondria to accommodate an increased thermogenic role for skeletal muscle (24). Together these studies suggest that cold exposure increases electron transport chain activity and mitochondrial density similarly in endothermic avian and mammalian systems. The necessity to use a UCP1 knockout mouse to better assess the role of skeletal muscle in thermogenesis recommends that birds, which lack brown adipose tissue, may be better suited models to understand the role of skeletal muscle thermogenesis in response to environmental or dietary stimuli.
H+ leak across the inner mitochondrial membrane uncouples the electron transport chain from oxidative phosphorylation, preventing the capture of energy as ATP and enhancing its release as heat. In rodent-based studies, heat production through H+ leak is primarily associated with the inducible thermogenesis observed in brown adipose tissue. However, the leak of H+ in liver and skeletal muscle mitochondria has been estimated to be responsible for 20% of basal energy expenditure in the rat (25). In fact, it accounts for 50% of respiration in perfused rat skeletal muscle (25). By comparing the respiration rate of mitochondria collected from rat and frog heart in the absence of ADP, Akhmerov showed that H+ ion leak was six- to sevenfold greater in isolated rat mitochondria than in isolated frog mitochondria (26). This may be in part explained by the inner mitochondrial membrane surface area per mitochondria being larger in mammals than in reptiles of a similar size (1). However, it may also be due to differences in the H+ gradient and membrane lipid composition.
Proton leak increases with mitochondrial membrane potential across species. Still, at the same mitochondrial membrane potential, H+ ion leak is approximately fivefold greater in the rat than in the bearded dragon (2). Other ectothermic species mute H+ leak by maintaining a lower mitochondrial membrane potential. In the rainbow trout, mitochondrial proton leak at a given membrane potential is higher than that observed in mitochondria from rat or pigeon (27). This is because membranes in cold-water fish species have a high degree of polyunsaturated fatty acyl groups to maintain membrane fluidity at low temperatures. The increase in polyunsaturation also increases ion permeability (28). To limit proton leak, the rainbow trout has low electron transport chain activity eliciting a low mitochondrial membrane potential. Accordingly, total proton leak is lower in the rainbow trout than in the endothermic rat and pigeon. Thus, proton leak can be manipulated by either altering electron transport chain activity and mitochondrial membrane potential or by changing H+ ion permeability.
Although proton leak is typically associated with heat production, increasing proton leak can also allow for metabolic flexibility. Accordingly, fish express UCP1, 2, and 3 (29, 30). In the carp, UCP1 is primarily expressed in the liver and expression is downregulated by a decrease in water temperature (30). The liver plays a central role in synthesizing and distributing glucose, ketones, and lipids to the rest of the body. Therefore, an increase in water and body temperature would increase the metabolic demand of non-hepatic tissues and necessitate expression of UCP1 to regenerate FAD+ and NAD+. In line with increased gluconeogenic demand, hepatic phosphoenolpyruvate carboxykinase (PEPCK) enzyme activity, essential for greater gluconeogenic flux, is increased in the Antarctic eelpout in response to a 5°C increase in water temperature (31). Thus, the direct relationship between temperature and UCP1 expression may encourage the regeneration of hepatic FAD+ and NAD+, essential for flux through β-oxidation to produce the acetyl-CoA necessary for ketogenesis while sparing amino acid oxidation. Accordingly, in the fish upregulation of UCP1 is not an adaptation to increase hepatic heat production, but instead to promote metabolic flexibility. Skeletal muscle primarily expresses UCP3 in both fish and rodents (30, 32). Interestingly, UCP3 is also similarly upregulated by fasting in both rodents and fish, while fasting increases proton leak from skeletal muscle mitochondria in the cane toad (15). As we proposed for UCP1 in liver, it has been proposed that this upregulation of UCP3-mediated H+ leak is essential for the increased reliance on fatty acid oxidation during a fast (32). Thus, studies of UCP regulation in fish can improve our understanding of UCP-mediated changes in metabolic flux without the demand for heat generation.
Conversely, some ectothermic fish species appear to manipulate H+ leak to warm-specific tissues above environmental temperature. Indeed, cold acclimation increases UCP1 expression in the optic tectum, a brain region important for sight, in the carp (33). This warms the brain’s optic center to maintain eyesight in cold environments, through a mechanism similar to the Ca++ cycling-based heater organ in deep diving marine species (34, 35). In sharks, skeletal muscle endothermy is thought to increase swimming rate, allow for thermal niche expansion, and increase digestive enzyme activity and digestive system passage (36). The mako shark uses red muscle fibers for endothermia. Mitochondria from mako shark red muscle fibers and liver display increased succinate stimulated respiration and maintain membrane potentials that exceed that of mitochondria from two ectothermic sharks. Although mitochondria from all of these shark species have similar levels of proton leak at a given membrane potential, the increased proton gradient in mako sharks drives H+ leak and heat production (37). Together these studies suggest that heat production through H+ leak may be used in some ectothermic species to warm specific tissues or maintain a set minimal body temperature required for survival.
In endotherms (birds and mammals), proton leak decreases with increasing body mass and explains 67% of the variability in standard metabolic rate (27, 38, 39). Proton leak is robust in birds. In fact, liver mitochondrial proton leak in all birds studied, including the much larger emu and goose, exceeded that of the rat (39). Across bird species, the ratio of monounsaturated fatty acids in phospholipids increases with body mass (39). The increase in monounsaturated fatty acyl groups may play a role in the decreased proton leak associated with increased body mass. In fact, membranes rich in monounsaturated fatty acyl groups are typical in ectothermic species with low levels of membrane ion permeability (40).
In rodents, studies aimed at understanding the endothermic response to cold have focused on the role of UCP1-mediated H+ ion leak in brown adipose tissue (41). Brown adipose tissue is integral to the maintenance of homeothermy in newborn infants (42–44). However, in humans, the passage from newborn through childhood and into adulthood results in a decrease in the amount of UCP in adipose tissue, suggestive of a reduction in metabolically active brown adipose tissue (45). The recent identification of brown adipose tissue depots near the clavicle in the adult human has spurred a resurgence in research aimed at better understanding the role of brown adipose tissue in energy expenditure of adult humans (46, 47). Yet the relative mass of brown adipose tissue in the adult human is minimal (168 ± 56 cm3). Accordingly, animal models that lack brown adipose tissue may provide a better understanding of the role of H+ leak in the thermogenesis of other tissues (48).
Birds, which lack brown adipose tissue, are uniquely suited to address the role of H+ leak in the endothermic response to environmental stimuli (cold or fasting). Avian UCP is expressed in skeletal muscle and increases in cold acclimated ducklings and penguins (49, 50). Proton leak across the inner mitochondrial membrane is higher in skeletal muscle from penguins that have been exposed repeatedly to cold water over 20 days. Teulier et al. observed that cold exposure of ducklings increased whole animal metabolic rate and ADP-stimulated oxidative phosphorylation. Surprisingly, despite the upregulation of avian UCP expression, there was no effect of cold acclimation on the efficiency of ATP synthesis or non-phosphorylating respiration. Accordingly, Teulier et al. propose that avian UCP primarily increases heat production by increasing aerobic skeletal muscle metabolic flux, rather than enhancing H+ leak (51).
In addition to the UCPs, the ANT may encourage proton leak through the inner mitochondrial membrane (52). Avian ANT protein expression increases in response to repeated cold water immersion (50). Therefore, both ANT and UCP are upregulated in cold acclimated birds and may be integral to the maintenance of body temperature through upregulated skeletal muscle thermogenesis. During a fast, the maintenance of body temperature may partially depend on mitochondrial H+ leak. Skeletal muscle avian UCP expression increases with glucagon treatment in ducklings (49). In the hummingbird, which undergoes torpor every night to endure an extended fast, skeletal muscle UCP expression increases during torpor and likely serves to help the bird re-warm from the torpid state (53). Thus, skeletal muscle is an integral site of mitochondrial proton leak, a key source of heat production in endotherms.
Non-mammalian vertebrates have been integral to understanding the thermogenic potential of Ca++ cycling. Both fish and birds use Ca++ cycling to generate body heat independent of locomotion. To understand the unique physiological adaptations that each have evolved, we must first understand the basis of the Ca++ cycle. Within skeletal muscle the sarco/endoplasmic reticulum Ca++ ATPase (SERCA) pumps Ca++ from the cytosol into the sarcoplasmic reticulum against the concentration gradient, while hydrolyzing an ATP to ADP and inorganic phosphate. The hydrolysis of ATP releases some energy as heat. The remainder of the energy is released as heat when Ca++ leaks from the sarcoplasmic reticulum into the cytosol down the electrochemical gradient (Figure 1). This cycle is essential for calcium oscillations in muscle that allow for contraction.
Marlin, swordfish, and tuna all have an endothermic “heater” organ that raises the temperature of the brain and eye (35, 54–56). This heater organ is a highly vascularized, modified eye muscle rich in mitochondria and cytochrome oxidase activity (35, 54–56). By maintaining the central nervous system and eyes at temperatures above ambient temperature, the heater organ improves eye sight and central nervous system function in cold ambient temperatures (35, 56–58). Warming the retina improves temporal resolution up to 10-fold, giving these deep diving, visual predators a crucial advantage over prey species (56). In addition to being mitochondria rich (68% of total cell volume), the heater organ contains a great deal of sarcoplasmic reticulum rich in SERCA1b and the ryanodine receptor 1 (34, 55). This encourages heat production from ATP hydrolysis as the SERCA pumps Ca++ into the sarcoplasmic reticulum and additional heat production as that Ca++ flows through the ryanodine receptor 1 down the concentration gradient back into the cytosol (34). The lack of contractile proteins uniquely positions the heater organ to take full advantage of heat generation by calcium cycling without motor consequence (58).
Cold exposure increases basal metabolic rate in birds, rodents, and humans (59–62). However, unlike rodents, which rely heavily on brown adipose tissue for thermogenesis, birds lack brown adipose tissue and rely more heavily on skeletal muscle thermogenesis. In the Muscovy duck, in vivo cold exposure increases ex vivo skeletal muscle oxygen consumption by 25% (63). Acute cold exposure of ducklings increases cardiac output and directs blood flow toward thermogenic sites, resulting in a 130% increase in skeletal muscle blood flow (64), indicating that skeletal muscle is the primary thermogenic organ in the duckling. In fact, birds rely on skeletal muscle calcium cycling to maintain body temperature at low environmental temperatures. Accordingly, acute 24-h cold-exposure increases skeletal muscle Ca++ ATPase activity (21). Cold acclimation over 5 weeks in ducklings increases sarcoplasmic reticulum SERCA1, SERCA2, and ryanodine receptor expression and activity as evidenced by increased Ca++ uptake and ryanodine binding (65, 66). The increase in Ca++ cycling leads to an increased resting metabolic rate, ATP demand, and oxygen consumption in cold acclimated birds (63, 67, 68). This is accommodated by increased expression of components of the electron transport chain, including cytochrome C and NADH ubiquinone oxidoreductase (21).
Interestingly, cold exposure stimulates lipolysis in birds, rodents, and humans (21, 62, 69). Mobilized lipids provide a source of carbons to meet this increased metabolic demand associated with maintaining body temperature. Accordingly, in the mouse, fatty acid translocase (cd36) knockout prevents the maintenance of body temperature in response to cold exposure (70). Moreover, cold acclimation in the sparrow increases pectoralis cd36 protein expression by 46% (71). In addition to acting as a carbon source, these lipids may modulate the degree of Ca++ cycling. Long-chain acyl carnitines accumulate in the skeletal muscle of cold acclimated ducklings. Palmitoyl (16 C) carnitine activates Ca++ release from duckling skeletal muscle sarcoplasmic reticulum (72). Thus, this skeletal muscle non-esterified fatty acid accumulation and the potential downstream consequences are conserved from birds to rodents and humans (62).
Data from the partially endothermic fish species and homeothermic avian species has established the thermogenic potential of skeletal muscle and the role of skeletal muscle calcium cycling in heat generation and body temperature maintenance. More recently, studies focused on sarcolipin (SLN), a protein that uncouples ATP hydrolysis from sarcoplasmic reticulum Ca++ sequestration, have elucidated a mechanism by which mammals use this calcium sequestration machinery to generate heat (73). Normally, SERCA hydrolyzes ATP and uses the majority of that energy to pump Ca++ from the cytoplasm against a concentration gradient into the sarcoplasmic reticulum, while releasing the remaining energy as heat. By decreasing Ca++ sequestration, SLN increases the heat released per mol of ATP hydrolyzed by more than 50% (74–76). The Periasamy laboratory has conducted a set of elegant studies to investigate the role of SLN in the response to thermogenic and dietary challenges (77, 78). They first showed that UCP1 knockout mice, which lack the thermogenic potential of brown adipose tissue, express more SLN in skeletal muscle (78). SLN overexpression in skeletal muscle increases oxygen consumption (77). This increase in skeletal muscle oxygen consumption translates to whole body energy metabolism, as SLN overexpressing mice lose weight when pair fed with wild-type mice. Furthermore, when challenged with a high fat diet, these SLN overexpressing mice eat more yet gain less body weight than wild-type mice (77). These findings in the mouse may translate to humans, as obesity alters methylation in the promoter of the ryanodine receptor 1 gene (79). In fact, this effect on the RYR1 promoter is the most prominent methylation response to obesity, most heavily affected. Moreover, the normal pattern of methylation can be restored with weight loss (79).
Fish and birds have been essential in establishing the role of calcium cycling and the SERCAs in energy expenditure and heat production. Deep water fish have developed a unique tissue with specified function to maintain brain and eye temperatures during exposure to deep cold ocean waters, while endothermic birds establish the potential for skeletal muscle to act as a thermogenic organ. New research reporting that calcium cycling is important for body weight regulation in rodents provides an exciting new avenue for drug development to combat obesity. Humans and birds both express higher levels of SLN than mice, recommending that birds may be a valuable model organism for assessing the role of SLN in body weight regulation.
Plasma Membrane Potential
The maintenance of the plasma membrane potential is a critical component of cellular homeostasis and requires the tight regulation of intracellular ion concentrations. Passive ion channels and active transport pumps establish Na+, K+, and Cl− electrochemical gradients across the plasma membrane that are essential for fundamental cellular processes. Transport of molecules both into and out of the cell, hormone secretion, muscle contraction, and neuronal communication are all dependent on the maintenance of resting membrane potential and utilization of these energetically favorable ion gradients. The Na+/K+ ATPase is the primary active pump driving cellular membrane potential (Figure 1).
Na+/K+ ATPase activity represents a major cellular energy demand and significant portion of resting metabolic activity. At the whole animal level, the Na+/K+ ATPase accounts for ~25% of ATP consumption in mammals (80). However, this can vary widely by tissue. In the liver, Na+/K+ ATPase activity constitutes ~10% of cellular energy use, while this rises to 60% in brain and kidney (81). Comparing the relative contribution of energy consuming processes between endotherms and ectotherms, 60 and 54% of cellular respiration is directed toward ATP production, of which ~13.3 and ~18.5% is consumed by the Na+/K+ ATPase in rat and lizard hepatocytes, respectively (2, 82). Since the respiration rate of rat hepatocytes is about four times that of lizard hepatocytes, 5.6 times more ATP is allocated toward Na+/K+ ATPase activity.
While endotherms and ectotherms maintain similar Na+/K+ ATPase densities across tissue type, the molecular activity of the pump is four to five times higher in endotherms (83). Accordingly, the plasma membrane passive permeability to both Na+ and K+ is four- to ninefold greater in endotherms than in ectotherms at the same temperature (84–86). Thus, to maintain established ion gradients, the leakier cell membranes of endotherms require more active Na+/K+ ATPase. Increased total ion flux in endotherms may be an evolutionary adaptation to increase heat production. The resulting higher activity of the Na+/K+ ATPase, in part, accounts for the greater level of energy expenditure in endotherms.
The lipid composition of the plasma membrane has a primary role in regulating Na+/K+ ATPase activity. Among ectotherms, the degree of membrane polyunsaturation is significantly correlated with Na+/K+ ATPase activity (87). In membrane crossover experiments, reconstitution of the Na+/K+ ATPase of an ectotherm (cane toad or crocodile) in the more polyunsaturated membrane of an endotherm (rat or cattle) increases pump activity by 40–180% while reconstitution in the reverse direction decreases Na+/K+ ATPase activity by 40–250% (88, 89). Thus, the inherent properties of the plasma membrane strongly regulate Na+/K+ ATPase activity. Interestingly, membranes of cold water fish have higher levels of unsaturated phospholipids than their warm water counterparts (90). In carp liver slices, exposure to decreasing temperatures immediately inhibits the synthesis of saturated fatty acids and stimulates desaturase activity (91). Across fish species, acclimation to low temperatures increases the degree of membrane fatty acyl unsaturation, stimulating Na+/K+ ATPase activity to compensate for the cold-induced decline in enzyme activity (92). Ectotherms adapt to temperature by changing membrane composition, representing a key regulatory mechanism by which activity of membrane-integrated enzymes, including the Na+/K+ ATPase, are altered to maintain membrane potential in the face of variable environmental conditions. Still, temperature changes have a more robust effect on active pump processes than passive ion leak, giving rise to the possibility of temperature disrupted ion gradients. In fact, in skeletal muscle of cane toads, bullfrogs, and black racer snakes, intracellular Na+ concentrations are higher at 20°C compared to 30°C, suggestive of decreased Na+/K+ ATPase activity with increasing environmental temperature (93). In cold-adapted species, activity of the Na+/K+ ATPase is less temperature sensitive, limiting temperature-dependent changes in membrane potential. In fact, the molecular activity of the Na+/K+ ATPase from an Antarctic octopus is 400% greater than that of the temperate octopus at 10°C (94).
Many ectotherms that overwinter underground or are exposed to prolonged water submersion have developed strategies to tolerate hypoxia. Turtles can spend over half of their life in an overwintering state and survive sustained periods of anoxia (95). In the western painted turtle hepatocyte, 28% of total cellular ATP is utilized by the Na+/K+ ATPase in normoxia (96). In response to anoxia, the Na+/K+ ATPase activity decreases by 75%, but accounts for nearly three-fourth of total cellular ATP turnover. Suppressed Na+/K+ ATPase activity is partially mediated by adenosine signaling which is robustly stimulated during anoxia (97). Ion gradients and plasma membrane potential are maintained during anoxia. Thus, passive ion flux must be downregulated in anoxia to match the decrease in active ion transport. In the turtle, neurons adapt to low oxygen by limiting K+ leak through a fivefold reduction in the open probability of Ca++ gated K+ channels (98). However, not all ectotherms can maintain ion gradients when challenged with low oxygen. In rainbow trout hepatocytes, an anoxia-intolerant species, cellular ATP content, and Na+/K+ ATPase activity are rapidly reduced by anoxia while K+ efflux rates exceed K+ influx up to eightfold (99). In the anoxia-tolerant goldfish, anoxia diminishes K+ efflux to match the decline in Na+/K+ ATPase activity, resulting in a net flux of K+ close to 0. Therefore, downregulating ion leak and allocating a greater percentage of total cellular ATP to the Na+/K+ ATPase appear to be principal strategies in maintaining membrane potential during metabolic arrest in anoxia-tolerant species.
This comparison between anoxia tolerant and intolerant fish species recommends that hypoxia-induced cellular damage may be limited by inhibiting K+ efflux channels within cells. Indeed, hypoxia induces K+ efflux in mammalian tissues (100–102). In fact, inhibition of K+ efflux across the plasma membrane prevents neuronal apoptosis in rats subjected to transient middle cerebral artery occlusion to induce hypoxia and ischemia or in serum-starved apoptosis-induced mouse neocortical neurons (103). In addition, the role of K+ efflux in hypoxia-induced apoptosis may be used to better understand the resistance of cancer to hypoxia-induced apoptosis. Several human cancers express low levels of K+ channels, preventing K+ efflux, and resulting in resistance to apoptosis (104). As in the anoxia tolerant goldfish, this diminished K+ efflux, allows cancer cells to maintain a nearly normal membrane potential in the face of depressed Na+/K+ ATPase activity. Thus, the strategies developed by anoxia tolerant species may be used to limit hypoxia induced tissue damage caused by cardiac arrest or arterial occlusion, while the sensitivity of anoxia intolerant species may provide insight into treatments that will diminish the hypoxia resistance observed in cancer.
Maintaining ion gradients demands at least 50% of mammalian resting metabolic energy expenditures. By adapting body temperature to the environment, ectotherms are not reliant on ion flux down the mitochondrial membrane H+, sarcoplasmic reticulum Ca++, or plasma membrane Na+/K+ gradients for internal heat generation. Because they are not as stringently dependent on the energetic costs of endogenous thermoregulation, ectotherms can manipulate these ion gradients to adapt to a wide range of environments and stressors. Like mammals, avian species are constrained by the demands of homeothermy. Yet, they lack the endothermic brown adipose tissue depots that constrain interpretation of rodent-based findings. In turn, having adapted to a range of environments, avian species provide an ideal comparative model to understand the regulation of ion flux to manipulate heat generation. The diversity of non-mammalian vertebrate species and environmental niches to which they have adapted provide unique insight into ion gradients that are lacking in studies of mammalian species. Ectothermic fish manipulate H+ and Ca++ leak to warm-specific tissues, providing ideal models to further investigate the regulation of ion leak. Avian species have been essential in establishing the importance of skeletal muscle mitochondrial density and Ca++ ATPase activity in thermogenesis. Recent work highlighting the role of SLN in mammalian body weight homeostasis validates the application of these avian studies to understand mammalian physiology. Finally, studies that compare the ability of anoxia tolerant and intolerant fish species to maintain plasma membrane potential may be applied to prevent the damage associated with stroke- or cardiac arrest-induced hypoxia and ischemia. In addition, these studies can be applied to exacerbate sensitivity to hypoxia in cancer. The vast number of lower vertebrate species and the varied environments to which they have adapted allow for unique research opportunities that can be exploited to expand our knowledge of mechanisms underlying metabolic flux, energy expenditure, and body weight regulation.
CG, KK, and BR all made substantial contributions to the conceptual framework of this review, participated in the drafting and revising of the manuscript, approved this final version for submission, and agreed to be accountable for all aspects of the work.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
This work is supported by the National Institute of Food and Agriculture, U.S. Department of Agriculture, under award number 2015-70007-24236 and by the U.S. Department of Agriculture Western Regional Aquaculture Center award entitled Efficient, rapid assay for predicting the growth rate of aquaculture species based on metabolic rate of the fertilized egg.
UCP, uncoupling protein; ANT, adenine nucleotide translocase; NADH, nicotinamine adenine dinucleotide; ATP, adenosine triphosphate; ADP, adenosine diphosphate.
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30. Jastroch M, Wuertz S, Kloas W, Klingenspor M. Uncoupling protein 1 in fish uncovers an ancient evolutionary history of mammalian nonshivering thermogenesis. Physiol Genomics (2005) 22:150–6. doi:10.1152/physiolgenomics.00070.2005
31. Windisch HS, Kathover R, Portner HO, Frickenhaus S, Lucassen M. Thermal acclimation in Antarctic fish: transcriptomic profiling of metabolic pathways. Am J Physiol Regul Integr Comp Physiol (2011) 301:R1453–66. doi:10.1152/ajpregu.00158.2011
32. Samec S, Seydoux J, Russell AP, Montani JP, Dulloo AG. Skeletal muscle heterogeneity in fasting-induced upregulation of genes encoding UCP2, UCP3, PPARgamma and key enzymes of lipid oxidation. Pflugers Arch (2002) 445:80–6. doi:10.1007/s00424-002-0879-9
33. Jastroch M, Buckingham JA, Helwig M, Klingenspor M, Brand MD. Functional characterisation of UCP1 in the common carp: uncoupling activity in liver mitochondria and cold-induced expression in the brain. J Comp Physiol B (2007) 177:743–52. doi:10.1007/s00360-007-0171-6
34. Morrissette JM, Franck JP, Block BA. Characterization of ryanodine receptor and Ca2+-ATPase isoforms in the thermogenic heater organ of blue marlin (Makaira nigricans). J Exp Biol (2003) 206:805–12. doi:10.1242/jeb.00158
36. Newton KC, Wraith J, Dickson KA. Digestive enzyme activities are higher in the shortfin mako shark, Isurus oxyrinchus, than in ectothermic sharks as a result of visceral endothermy. Fish Physiol Biochem (2015) 41:887–98. doi:10.1007/s10695-015-0055-8
37. Duong CA, Sepulveda CA, Graham JB, Dickson KA. Mitochondrial proton leak rates in the slow, oxidative myotomal muscle and liver of the endothermic shortfin mako shark (Isurus oxyrinchus) and the ectothermic blue shark (Prionace glauca) and leopard shark (Triakis semifasciata). J Exp Biol (2006) 209:2678–85. doi:10.1242/jeb.02317
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49. Raimbault S, Dridi S, Denjean F, Lachuer J, Couplan E, Bouillaud F, et al. An uncoupling protein homologue putatively involved in facultative muscle thermogenesis in birds. Biochem J (2001) 353:441–4. doi:10.1042/bj3530441
50. Talbot DA, Duchamp C, Rey B, Hanuise N, Rouanet JL, Sibille B, et al. Uncoupling protein and ATP/ADP carrier increase mitochondrial proton conductance after cold adaptation of king penguins. J Physiol (2004) 558:123–35. doi:10.1113/jphysiol.2004.063768
51. Teulier L, Rouanet JL, Letexier D, Romestaing C, Belouze M, Rey B, et al. Cold-acclimation-induced non-shivering thermogenesis in birds is associated with upregulation of avian UCP but not with innate uncoupling or altered ATP efficiency. J Exp Biol (2010) 213:2476–82. doi:10.1242/jeb.043489
52. Brand MD, Pakay JL, Ocloo A, Kokoszka J, Wallace DC, Brookes PS, et al. The basal proton conductance of mitochondria depends on adenine nucleotide translocase content. Biochem J (2005) 392:353–62. doi:10.1042/BJ20050890
67. Filali-Zegzouti Y, Abdelmelek H, Rouanet JL, Cottet-Emard JM, Pequignot JM, Barre H. Involvement of the catecholaminergic system in glucagon-induced thermogenesis in Muscovy ducklings (Cairina moschata). Pflugers Arch (2000) 441:275–80. doi:10.1007/s004240000409
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Keywords: ectotherm, endotherm, energy expenditure, membrane potential, mitochondrial membrane potential, H+ gradient, Ca++ gradient, Na+/K+ gradient
Citation: Geisler CE, Kentch KP and Renquist BJ (2017) Non-Mammalian Vertebrates: Distinct Models to Assess the Role of Ion Gradients in Energy Expenditure. Front. Endocrinol. 8:224. doi: 10.3389/fendo.2017.00224
Received: 30 January 2017; Accepted: 18 August 2017;
Published: 01 September 2017
Edited by:Maximilian Michel, University of Michigan, United States
Reviewed by:Elisabeth Eppler, University of Basel, Switzerland
Jon Ramsey, University of California, Davis, United States
Copyright: © 2017 Geisler, Kentch and Renquist. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Benjamin J. Renquist, firstname.lastname@example.org


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AFCON 2023: Cape Verde vs South Africa: Predicted lineup, injury news, head-to-head, telecast
South Africa can qualify for semis for the first time after 2000
One of the surprises of the AFCON, Cape Verde have defined all the odds to top the group consisting of Egypt and Ghana. Very few expected Cape Verde to outperform them to top the group. Cape Verde defeated Mauritania by 1-0 in the round of 16 to qualify for the quarter-finals thanks to Ryan Mendes converting the penalty.
With their performance against last year’s semi-finalist Morocco, South Africa shocked the world by knocking them out of the AFCON in the round of 16. Makgopa and Mokoena scored in the second half to advance to the quarter-final. For the first time since 2000, South Africa will feel confident that they can make it to the semi-finals after the top five favourites of the tournament have already been eliminated.
Sun, 4 February 2024 at 1:30 AM IST
Location: Stade Charles Konan Banny de Yamoussoukro
Cape Verde (in all competitions): W-D-W-W-L
South Africa (in all competitions): W-D-W-L-D
Player to Watch
Ryan Mendes (Cape Verde)
A crucial component of Cape Verde’s road to the quarter-final has been Ryan Mendes. He has scored two goals and provided one assist. His decisive goal against Mauritania in the round of 16 earned them a place in the quarter-final. In addition to having a clear eye for goal, Ryan Mendes is capable of creating team chances in the final third.
Themba Zwane (South Africa)
The 34-year-old forward has been instrumental in the success of South Africa this term as he assisted in the last match against Morocco to send them into the quarter-final and also scored a brace against Namibia in the group stage. In addition to his excellent hold-up play, he has also been an excellent team player when bringing teammates into the game.
Trending World Football Articles:
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- Cape Verde are unbeaten in the last four matches
- South Africa defeated Morocco in the last match by 0-1
- Cape Verde have won the previous two matches against South Africa
Injury and Team News
There is a fully fit squad for Cape Verde to choose from. For the first time since 2000, Hugo Broos will likely stick to the same eleven that defeated Moroccan in the last match. After a famous win over Morocco, Hugo Broos is likely to stick with the same eleven.
Head to Head
Cape Verde Predicted lineup (4-3-3):
Vozinha; Moreira, Costa, Lopes, Paulo; Monteiro, Lenini, Duarte; Mendes, Bebé, Cabral
South Africa Predicted lineup (4-2-3-1):
Williams; Mudau, Kekana, Mvala, Modiba; Mokoena, Sithole; Morena, Zwane,Tau; Makgopa
Match Prediction for Cape Verde vs South Africa
With both teams playing on counterattacks, small margins will decide this match. Because Cape Verde beat South Africa twice in the past, they will be confident heading into this match. On the other hand, South Africa are confident after defeating Morocco in their last match.
Prediction: Cape Verde 1-2 South Africa
Telecast for Cape Verde vs South Africa
UK – Sky Sports
US – beIN Sports
Nigeria – NTA
AFCON 2023 Worldwide Broadcast Details: Click here
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domain: sports


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oday I am going to present one of my favourites MMORPG games - Dekaron.
The game has been released by the European games distributor Gametribe and is now developed by NexonEU.
It started as a small multiplayer game with 5 main classes(Vicious Summoner, Azure Knight, Bagi Warrior, Incar Magician, Segita Hunter). In time it has gained millions of players and also released another class - The Aloken.
The game is 3D and takes place in a fantasy world, where humanity and its protectors fight the evil forces.
The controls are easy to understand, and new players can find a step-by-step tutorial.
Choosing the character doesn't give you many choices. The classes can not be customized, which is a negative point for a game nowadays.
Skills and stats are basic-you gain a skill point and five stat points per level. Currently the max level is 200, though there is no player who has ever reached it yet. Every class has its own abilities, and its own way to use the stat points ( For example, a Bagi Warrior needs more points in Strengh, but an Incar Magician needs them in Magic Atack) .
The graphics aren't made for the young generation of gamers. The details aren't that good, and you can actually move through different objects.
Even though the graphics aren't a positive point, Dekaron still impresses with its action. There are many maps where you can level, many monsters to kill, and many dungeons to do. Although, a person who gets bored fast might not like it since you have to spend some time on leveling.
The game is currently expecting a new update, that will include two new classes, new events, armors and weapons.
The Nexon Staff is friendly and active. Like every multiplayer game, Dekaron has its hackers, but the GMs are working hard on keeping the game clean.
To get the game, you have to follow a few steps :
- Go to nexoneu.com
- Create a free account
- Choose Dekaron from the game list
- Download it!
Simple as that! I hope you liked the review, looking forward to your replies.

domain: game


text:
While our support for religious freedom abroad might have limited resultants, we must remain vigilant at home.
‘Zealot’ author Reza Aslan on MSNBC Nobody has ever mistaken me for a Muslim, but I now feel an affinity … Continued
I am not now, nor have I ever been, a member of the Catholic Church. That’s why I feel less … Continued
When I was six, I began Hebrew School as an Orthodox Jew because that’s what my family was and that’s … Continued
There were two categories of teenagers in the 1950s: those who could name one book by an atheist and those … Continued
(Streeter Lecka — GETTY IMAGES) I could not have had a more patriotic beginning. I was born on Flag Day … Continued
can those who are “good without a god” really enter the gates of heaven? Popes ponder whether or not religious belief is crucial to salvation.
Recently I was invited to participate in a Religion and Law Conference at Florida State University. Almost all the other … Continued
DAVID MANNING REUTERS Eagle Scout Zach Wahls, 20, of Iowa City, Ia., talks after delivering a 280,000 signature petition to … Continued

domain: news


text:
Victims targeted by fraudsters lose "much more" than money, and that is why sentencing needs to be tougher, a high ranking Judge has said.
Sentencing Council chairman Lord Justice Treacy explained:
Fraudsters are in it to make money, but for their victims it can mean much more than losing money.
Our research with victims showed the great impact it can have on them, so the guideline puts this impact at the centre of considerations of what sentence the offender should get.
More top news
Passengers cry out and prayers are heard as a plane endures turbulence so severe it forced the pilot to take an unscheduled landing.
Michael Swain says the 'fantastic' surgery has made a huge difference to his life.
The brother of a teen who disappeared in 1979 has told ITV News he is "convinced" he was a victim of an alleged Westminster paedophile ring.


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WARNING JAYA: Tony Pua has destroyed Najib Razak's claim that he and Finance Minister Lim Guan Eng are opposed to one another by the decision of the credit rating agency to reduce their Petron's perspective.
Pua, who is Political Secretary at Lim, said that the former Prime Minister Najib's claim is unfounded. He explained that he had said that the revised Moody's Investors Service's prospects for Petron had not been downgraded.
"Lima cited the (outlook shift) from stable to negative, but I refer to the fact that, while there were changes in prospects, rating level A1 remained unchanged.
"There's nothing controversial … I do not care what Najib wants to say, because he is no longer relevant to our community," Pua said here about the housing conference.
Pua spoke earlier on the 2019 budget conference, Wisma Rehda, whose main organization is Rehda Real Estate and Real Estate Developers Association.
Vajd Najibs, who is also a former finance minister, went to Facebook to point out the apparent controversy at Lim and Pua stands on this issue, referring to BFM's 89.9 interview, where Lim did not coincide with Petron's "downgrade".
It followed Putrajaya's announcement that Petron would cost RM26 billion in dividends this year and will give RM54 billion in 2019 to the government.
Moody's Investors Service last week approved the Petrona A1 domestic issuer and foreign currency senior unsecured rating, but changed its outlook from stable to negative.
The negative outlook reflected the agency's view that Petron's financial position could deteriorate if Putrajaya continues to ask for high dividend payments.
Pua has previously said that Moody's is likely to be worried that the special dividend payments from Petron would be more than a one-off payment.
"A statement by a credit rating agency is that, if the government continues to take money, for example, from Petron, then it will be lowered," reports Pua last week.
Najib had invented both, "not on one page", saying that they were unlikely to have their own WhatsApp groups.

domain: finance


text:
Lucy Liu and Her Son, Rockwell, Make One Seriously Cute Duo
Lucy Liu has one serious cutie on her hands! Ever since the Elementary star welcomed her first child, a son named Rockwell Lloyd Liu, via surrogate back in August, the actress has been taking to Instagram to document her new life as a mom — and from the looks of it, she sure is enjoying it. Not only did the actress introduce her little bundle of joy to the world on social media, but Lucy's also treated her followers to snaps of Rockwell's nap times, his adorable outfits, and his precious smile. We've rounded up the duo's sweetest moments together, so check them out, then look at 52 families you should be following on Instagram.

domain: entertainment


text:
In Salavan province, the SFE works in five health centres. These centres enable villagers to receive basic health care as close to their village as possible.
A health centre usually includes an vaccinaton service, follows up pregnant women until they give birth, and offers consultations for common illnesses and care for minor wounds. In most cases, it also has a small pharmacy.
Our work among health centers aims to improve the quality of care, in particular through the training of health professionals as well as the regular follow-up of patients.
Every month, a team from the SFE visits the health centres joined by a doctor and a nurse from the provincial hospital. Depending on the needs, they provide training in bedside care, monitoring young mothers and infants, and diagnosing and treating common diseases.
Together, they take up the consultation logs and review which diagnoses were made based on which clinical signs and whether the proposed treatments were appropriate. It is an excellent practical exercise to improve the reception, care, and follow-up of people.
The health centres have no medical laboratory, radiology or ultrasound service. As a result, many pregnant women cannot go to the hospital for an ultrasound scan, so the ultrasound machine comes to them! Indeed, this examination is essential to know the number of babies and their state of health, their position and that of the placenta, etc. This information makes it possible to advise mothers on the place best suited for future childbirth.
Through monthly evaluations, we support each centre in improving its storage system and overall hygiene. Following a needs assessment, we provide medical equipment, storage cabinets or medical waste bins.
In some health centres, the SFE also offers renovation work. For example, one of the centres was built by villagers and had a concrete floor, which was very dusty in the hot season, and tin roofs, which were extremely noisy when it rained. The SFE installed a proper ceiling and tiled floors. The centre is now more welcoming, and more comfortable while better meeting hygiene criteria.
In one year of these activities in the health centres, the cleanliness and quality of care have improved significantly. In addition, as health workers sometimes feel isolated and under-resourced, they are grateful for the support, more resources, training and regular visits.
The entire SFE team is grateful to be able to contribute to the continuous improvement of health services in southern Laos.

domain: medical


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MONTPELIER, Vt. — Madeleine Kunin, 51, is Vermont's first woman governor and only the third Democrat to preside over the state since the Civil War.
If the mayor of Burlington, Vermont's largest city, has his way, Kunin's successor will be America's first Socialist governor.
"Vermont is ripe for a third-party takeover, and I may be the one to pull it off," said Bernard (Bernie) Sanders, 43, a Socialist, re-elected last month to his third term as mayor of Burlington.
Both Kunin and Sanders attribute their political success to the independent nature of Vermont voters. Both are Flatlanders, as Vermonters call residents not native of the state.
The low-key governor and the flamboyant mayor were born and raised outside Vermont. They both moved here when they were in their mid-20s.
Madeleine May was 24, a recent graduate of the Columbia School of Journalism, when she moved to Vermont in 1957.
"I didn't know anyone in Vermont. But I moved here because of an opportunity to work for the Burlington Free Press as a general assignment reporter," the governor said during an interview in her Capitol office.
"Most newspapers at the time would hire women only to do society news and write for women's pages. I wanted to cover all types of breaking news."
Paris in Her Plans
Her plans were to work in Burlington two or three years, then go to Paris and get a job as a reporter for the Paris Herald Tribune.
She never made it to Paris.
She stayed in Burlington. Married. Raised four children.
Twenty-eight years ago she came here as a stranger. Today everyone in Vermont (population 515,000) knows who she is.
Kunin was inaugurated Jan. 10 as Vermont's 74th governor. She is one of only two women governors currently in office; the other, Martha L. Collins, 48, is the Democratic governor of Kentucky. Unlike other governors, Kunin can never become President because she wasn't born in the United States.
In 1940, when Hitler's troops were marching through France, the governor's mother, a widow, fearing the Nazis' next invasion might be her homeland, Switzerland, packed up her belongings and sailed to America with her two children, Madeleine, 6, and Edgar, 10. Their father, a shoe importer, had died in a drowning accident in Switzerland when she was 3.
The three spoke German and knew no English when they arrived in America. To make ends meet, Kunin's mother supported herself and her two children baby-sitting, sewing and taking in boarders.
In her inaugural speech at the granite Doric-style 1859 Vermont Statehouse, with a gold dome topped by a statue of the Roman goddess Ceres, Kunin recalled her heritage:
"It is the immigrant spirit of hope which I wish to bring to state government. A spirit which instills in our children the belief that anyone can achieve anything in this country with hard work and education and a fair chance.
"I stand here with my husband and children. I stand here with the memory of my mother, my aunt, my grandmother who could never have dreamt I would be in this place on this day, but, who, through the courage of their lives, gave me the stamina to stand as tall as they did."
She paid tribute to the women of Vermont who made her election possible: "As first woman to take the oath of office of governor of the state of Vermont, I recognize that I was able to raise my right hand before you only because so many women had raised their voices, long before my words were spoken."
As a high school student, she planned to become a teacher. Under her picture in her Pittsfield, Mass., high school yearbook is a rhyme: "Mady will teach the ABCs, so class, may she have your attention, please?"
Decided on Journalism
But in college she decided on journalism. She has a BA in history from the University of Massachusetts, where she worked as a waitress to pay her way. She has an MA from Columbia and later earned another MA in English literature from the University of Vermont.
The 5-foot-7, trim, friendly, easy-going governor is married to Dr. Arthur Kunin, a kidney specialist and University of Vermont professor of medicine. They have three sons, ages 22, 19 and 15, and a 23-year-old daughter. Their youngest son lives at home, the two older boys are in college and their daughter is a sculptor's assistant.
Madeleine Kunin was one of the very few women elected to major political offices in 1984. A Democrat in a traditionally Republican state, she defeated Republican state Atty. Gen. John Easton by just 62 votes, more than the 50% that Vermont law requires for the $60,000 job as governor.
Kunin had been a typical middle-class homemaker, raising her family, before deciding to run for the state House of Representatives. The year was 1972. She won, served three two-year terms and became party whip and chairman of the Appropriations Committee. In 1978 she was elected to a two-year term as lieutenant governor and was reelected in 1980. In 1982 she ran for governor and lost. She ran again last year, this time winning.


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“This initiative gives those phones a second life and people in very traumatic situations some much-needed assistance.”
“You can’t help but be shocked by the scale of what ends up in our environment as litter,” Clean Up Australia Day 2024 date set.
“The dogs have an incredible sense of smell and can detect a single specimen of this weed in a huge area." Sniffer dogs making history.
Embracing the festive spirit, Cessnock Hospitality Group (CHG) has partnered with Samaritans for its 2023 Christmas Gift Appeal.
Three men have been charged with numerous offences, including assaulting police, following an incident at Lake Macquarie overnight.
Looking for a family-friendly event to catch the festive spirit this Christmas? Then look no further than Lake Macquarie.
HVO has used the feedback from submissions on its Environmental Impact Statement (EIS) to improve its proposal to continue mining until 2050.


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Lost in the Desert
I used this scene to learn and practice my 3D and interactive skills. It was inspired by some concept art done by a friend – I like the quirky yet eerie nature of this little lost station wagon deep in the desert. This was the first scene I created from scratch in Cinema4D.
I then took the car model and imported it into Unity to give it some interactivity using a tutorial. In the future I’d like to flesh out the interactive portions more, and have ideas for how to create a little mini multi-player game of dueling family station wagons lost in the desert.

domain: artistic


text:
‘Leading personalities being roped in to help in campaign to educate people about ill effects of tobacco’
Srinagar, May 31: Tobacco smoking at Public places in Jammu and Kashmir is on decline even as leading personalities of J&K are being roped in to help in campaigns to educate people about ill effects of tobacco smoking, Secretary Health and Medical Education Bhupinder Kumar said on Wednesday.
Talking to reporters at the side-lines of an event regarding “World No Tobacco Day” at Dental college Srinagar, Kumar said that this day is being observed to educate people about the ill effects of tobacco consumption and how to work jointly to decline the percentage of tobacco smoking and smokeless consumption”, he said.
“Every year this campaign is being run and laws are in place due to which tobacco smoking in open is on decline in J&K,” he said, adding, “Tobacco continues to remain a risk factor for deadly diseases like cancer and there is need to work jointly to fight this menace”.
“We have started a campaign in this regard and leading personalities like Dr Shahid Iqbal Choudhary, Dr Sushil Razdan, Parvaiz Rasool are being roped in as brand ambassadors to be part of campaign of educating people about ill effects of tobacco so as to decrease its percentage and keep young generation away from it so as to develop a healthy society,” he added.
He said that due to government efforts smoking in public places and women folk is on decline in J&K.
“All arrangements are being made to provide better facilities to Yatris,” Kumar said.
Around 3000 vaccines in the health department have been already being referred to concerned agencies and among them around 2000 have been filled and it is a continuous process and all vaccines will be filled,” Health and Medical Education Secretary Said.
Doctors who spoke during the event said that smoking is a gateway of drug abuse as almost all drug abusers start from smoking and then he is lured for drugs easily.
They said that 50 percent families get affected by second hand smoking, that is smoke that comes from the burning of a tobacco product and smoke that is exhaled by smokers and more importantly third hand smoking is the main concern which means smoking is residual nicotine and other chemicals left on indoor surfaces by tobacco smoke.
“People are exposed to these chemicals by touching contaminated surfaces or breathing in the off-gassing from these surfaces,” they said, adding that “Cigarette contain around 4,000 chemicals which include 200 are hazardous and around 60 cause cancers and smoking is known for causing multi organ cancers.”
They said that parents and teachers have to play a role in this regard to keep children away from smoking. “Taking of e-cigarettes is more hazardous and unregulated sale of imported cigarettes is there without labelling and branding with no hazardous symbols and signs,” they said.
“Multi-pronged strategy is required to reduce cigarette consumption and its ill effects,” they added. Meanwhile, MD NHM Ayushi Sudan on the occasion appreciated the health department for taking such initiatives and she was impressed with the idea of naming brand ambassadors as according to her more and more people need to be involved to carry forward the message of ill effects of tobacco consumption.
She said that the most important part of tobacco control is awareness and there is need to educate more and more people about its harmful effects and there is need to work in tandem to reduce the prevalence of tobacco consumption.
On the occasion Dr Shahid Iqbal Choudhary and Parvaiz Rasool who were present on the occasion and Dr Razdan in video message vowed to fight for tobacco control measures in J&K and hoped that their involvement will help in reduction in the number of tobacco users in J&K. They appealed people to quit smoking as tobacco is cause of many cancers—(KNO)

domain: sociology


In the face of a systemic homelessness that has economic, social, public health, cultural, spiritual, and ecological manifestations, how might we develop more holistic and integrated policies and practices for deeper homefulness in our communities?
Developed by Empire Remixed in partnership with the Sorrento Centre—co-hosts of the national Beyond Housing to Homefulness Symposium—this podcast seeks to deepen the discussion of home-making and housing in the Canadian context and beyond.
Our guests come from a variety of backgrounds—developers and architects, faith leaders and theologians, community developers, activists, and politicians.
Subscribe now on your favourite platform.
One Response to “Homefulness Podcast”
Congratulations. An excellent vision. I look forward to hearing more in future podcasts


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To support interprofessional care during COVID-19, until further notice, doctors can submit 14029 for chronic disease management visits provided via telehealth by an allied care provider. The change is effective June 1, 2020 and doctors may bill 14029 retroactively. Additionally, new clarifications are now available on the appropriate use of fee codes 14076 and 13706 (the family physician-patient telephone management fee and the family physician delegated patient telehealth management fee, respectively). Please see below for details on these fee code updates.
1. Billing changes to 14029 allow for telehealth visits
Until further notice, fee code 14029, chronic disease management by an allied care provider, can now be billed for telehealth visits. 14029 may be billed as one of the visits required for the billing of chronic disease management fee codes, read more here.
This change is effective as of June 1, 2020 and physicians may bill retroactively to this date. If you are billing for a visit more than 90 days prior to your submission, please submit such claims with submission code ‘A’.
2. Clarifying the use of 14076 and 13706
GPSC heard from you that further clarity is needed on the appropriate use of 14076 and 13706. Family physicians who meet the requirements of 14076 should continue to bill 14076, family physician-patient telephone management fee. Alternatively, this fee may be billed when the visit is delegated to a College-certified allied care provider employed by the physician practice. 14076 has a limit of 1500 payable visits per calendar year.
13706, family physician delegated patient telehealth management fee, was introduced as a temporary fee code in March 2020 and is intended to be billed by family physicians who do not meet the requirements of 14076 or have exceeded the annual billing limit of 14076.
Unlike 14076, 13706 cannot be billed in the following situations:
- 13706 cannot be billed as one of the two required visits for 14050-53, chronic disease management fees. Physicians who have had their 14050-53 billings rejected as a result may resubmit visit as 14076.
- 13706 cannot be billed on the same day as 14077, 14018, 14050-53, 13005 by the same physician for the same patient. Physicians who have had their visit rejected as a result should resubmit visit as 14076.


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ANDERSSON BELL X ASICS GEL-SONOMA 15-50 - BRIGHT ROSE / EVERGREEN
$ 100.00 was $ 140.00
This size is currently sold out
For the third time, Japanese footwear giant Asics is reconnecting with South Korean label Andersson Bell to drop a new selection of sneakers. The ‘Asics World’ ensemble draws inspiration from a fantasy world, where humans and mythical horned animals co-exist. At the centre of this world are the characters Frocto, Fissit-tas and Conni, who inspire the playful colourways featured in this bold collection. For this collaboration, ASICS and Andersson Bell reimagine the trail-ready GEL-SONOMA 15-50s. Mesh and leather panels pay tribute to the original model, which have been upgraded with technical laces, co-branded accents and flashes of neon.
Color: Bright Rose / Evergreen
Limit 1 per customer
***Limited Release styles are NOT eligible for a refund, return or cancellation after order is placed.***

domain: design


The singer-songwriter became engaged to girlfriend Georgina Gordon earlier this month after popping the question during a romantic cruise down The Thames.
Speaking on ITV's Lorraine today, Lucy said wedding plans were now in full swing: "We are trying to look at some castles up North. I want to wear a suit of armour but I'm not allowed!"
Instead, she'll probably settle for tartan.
"My family are Scottish so I think I'm going to be wearing a kilt. Hunting Stuart is my tartan. It will be great!"
Romantic Lucy also spoke about the engagement, saying she had been planning it for ages – but Georgina nearly beat her to the big moment!
"About a week before, I had been planning it for ages, and about a week before she went, 'I might ask you to marry me' and I was like 'don't, don't do that!'"
Lucky Georgina didn't because she would have missed out on Lucy's amazing engagement surprise.
"I had planned to get a boat to take us up the Thames and as we went under Albert Bridge - she's from the Wirral and all her family and friends had come down from Liverpool, they were standing on Albert Bridge so I said 'who's up there with the sparklers?' and when she looked she started crying and I just said 'will you marry me?'"
For exclusive competitions and amazing prizes visit Reveal's Facebook page NOW!


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Jeff Marrs, MD
To view more patient reviews and ratings for Dr. Marrs, please click below.
Hip, Joint Replacement, & Knee
Dr. Marrs is an orthopedic surgeon who specializes in hip, joint replacement, and knee. Dr. Marrs began practicing at Black Hills Orthopedic & Spine Center in 2002.
After completing his Bachelor of Science degree in computer information systems and business administration at Regis University in Denver, Colorado, he went on to complete his premedical requirements from the University of Colorado in Colorado Springs, Colorado.
Dr. Marrs then received his Doctor of Medicine degree at the Mayo Foundation for Medical Education and Research in Rochester, Minnesota. Dr. Marrs then underwent a residency in the Department of Orthopedics at the Mayo Foundation for Medical Education and Research in Rochester, Minnesota.
Dr. Marrs is certified by the American Board of Orthopaedic Surgery.
When he is not seeing patients, Dr. Marrs enjoys spending time with his family, golfing, fishing, and other outdoor activities.
Regis University, Denver, CO, 1987
University of Colorado, Colorado Springs, CO, 1992
Mayo Foundation for Medical Education and Research, Rochester, MN, 1997
Orthopedics, Mayo Foundation for Medical Education and Research, Rochester, MN, 2002
American Board of Orthopaedic Surgery
- Black Hills Surgical Hospital
- Total hip replacement
- Hip resurfacing
- Total knee replacement
- Total knee revision
- Total shoulder replacement
- Carpal tunnel release
- Anterior, posterior, anterolateral less-invasive procedures
- Total hip revision
- Partial knee replacement
- Total shoulder revision
- Knee arthroscopy
- "TKA in Charcot Joints" Mayo Clinic Department of Orthopedic Surgery, with B. Morrey and J. Parvizi (Accepted for Publication)
- "Identification of a Dual Leucine Zipper Kinase (DLK) involved in Rat Fracture Repair" Mayo Clinic Department of Orthopedic Surgery, Laboratory of Mark E. Bolander, M.D.
- "Photochemistry of Phosphorus Containing Compounds" University of Colorado, Colorado Springs, Laboratory of David R. Anderson, Ph.D.
Phone: (605) 341-1414
Office: Rapid City, SD


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Doing ~ Tasting Events
By Megan Dressler, Student Writer
Chef Mark Molinaro of the School of Hotel and Restaurant Management (SHRM) has his students host four tasting events throughout October and November in order to show off their cooking skills to the NAU community. The events help students gain hands-on experience in creating dishes from scratch and building a brand based around the dish.
The first of these events, called “Savvy Soups,” was held on October 6, 2014 in SHRM’s Marriott Demonstration Kitchen. The event was attended by members of the NAU faculty and staff, as well as by NAU President Rita Cheng.
[Photo caption: Savvy Soups Tasting lures NAU President Rita Cheng and husband, Tom Cheng, to sample the savory delights.]
Students turned their cooking stations into mini restaurants, theming their station around their soup. For example, one group chose Clam Chowder, so they dressed as sailors and built a New England-themed setting. Each group developed a different back story for their restaurant, a uniform to wear, and literature to distribute. The remaining tasting events are “Meat Madness” followed by “Pastapalooza” and the semester rounds out with dessert tasting at “Sweet Endings.”
Chef Molinaro is pleased with the impact that these events have on the students, observing that it not only allows them to learn how to cook beyond following a recipe, but also how to build a brand, create marketing materials, and, as he says, “use hospitality-related skills that they’re learning.”
One student called the series of events “Awesome,” while student Juliet Hilinski said, “We’re having fun. It’s a lot of fun!” The students are excited to show off their skills while doing what they love: serving dishes that they are proud of while creating a hospitable and inviting environment for their guests.
Categories: flagstaff business news nau business news students fall 2014 2014 tasting events savvy soups


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DS9Designs Tatting and Beads
Maple Leaf Pattern
Requires 2 shuttles, 2 colors of thread. When tatted with size 20 thread, the maple leaf measures about 3 1/4" x 3 1/4".
The pattern has written and diagrammed instructions for shuttle tatting.
Techniques Used: Zig zag chain, self-closing mock ring, shoe lace trick, rings and chains.
Pattern Price: $2.50


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USGS: Groundwater Influences Key to Sustainable Water Supply
White Pine County, Nev., is a critical recharge area for several major regional flow systems that extend north to the Great Salt Lake and south to the Colorado River, according to the U.S. Geological Survey study, "Water resources of the Basin and Range carbonate-rock aquifer system, White Pine County, Nevada, and adjacent areas in Nevada and Utah."
The groundwater systems underlying many of the valleys in eastern Nevada and western Utah are not isolated but rather contribute or receive flow from adjoining basins. Some large-volume springs cannot be supported entirely by the local recharge from the adjacent mountains and depend on water from potentially hundreds of miles away. The imbalance between recharge and discharge within a basin is an important factor in making decisions that could affect water sustainability in surrounding areas.
This report is the final product of the Basin and Range carbonate-rock aquifer system study ordered by Congress in the Lincoln County Conservation, Recreation, and Development Act of 2004. The report was prepared in collaboration with the Desert Research Institute and the state of Utah, and in cooperation with the Bureau of Land Management.
The USGS study included a water-resources assessment that analyzed the geologic framework and hydrologic processes influencing the quantity and quality of groundwater resources. This information is critical for water-resource agencies to make informed decisions about future water-supply issues. Understanding the factors that influence flows from the aquifer system is necessary in evaluating how future groundwater development may affect discharge, ecosystems, and water supplies. Knowing what factors influence the quantity and quality of groundwater resources is essential when developing management plans for a sustainable water resource.
The study designates basin and regional groundwater "budgets," for 13 hydrographic areas and the entire study area in White Pine County, Nev. Water budgets enable an accounting of water as it moves through Earth's atmosphere, land surface, and subsurface. In order to do this, scientists from the USGS and the Desert Research Institute studied the hydrogeology, recharge and discharge, groundwater flow and geochemistry of the aquifer system.
The study-wide average annual groundwater recharge exceeded annual discharge by about 90,000 acre-feet. Most of this groundwater surplus exits the study area through Snake Valley to the northeast or White River Valley to the south. Annual recharge exceeded discharge in Steptoe Valley by 53,000 acre-feet, and a significant amount of groundwater flows from this valley to adjacent areas such as Jakes and White River Valleys to the west and Spring and Lake Valleys to the east. Annual discharge exceeded recharge in White River Valley by 41,000 acre-feet, and a significant amount of groundwater flows into this valley from adjacent areas, such as Jakes, Steptoe, and Cave valleys.
Groundwater flows through three types of aquifers in White Pine County: a shallow basin-fill aquifer, a deeper volcanic-rock aquifer, and an underlying carbonate-rock aquifer. The basin-fill aquifer is the principal source of domestic and agricultural water supply, which is safe for human consumption.

domain: geography


The Prescription Drug Monitoring Program (PDMP) was once again proposed in the Missouri House of Representatives and the Missouri Senate. Fortunately it failed. The main bill that went the farthest was HB90. The proponents claim that this program is needed to stop doctor shopping. What they don’t tell you is that in 2015, doctor shopping was responsible for only 2.3% of all misused prescriptions for pain medicine. 85% of the people who misused prescriptions for pain medicine got the prescription from one doctor or got it from a friend or relative who got it from one doctor. It also violates Article I, Section 15 of the Missouri Constitution because it is a mandatory, involuntary database. The ultimate goal is a national prescription database. These bills should never become law. Sen. Claire McCaskill sponsored an amendment in the Comprehensive Addiction and Recovery Act which allows local PDMPs to apply for federal grants. It has been adopted in St. Louis County, Jackson County (Kansas City), the city of Columbia and other cities and counties in Missouri.
*** LOCAL UPDATE ***
The PDMP was on the agenda at the June 6, 2017 Community Involvement Committee meeting. They plan to bring it to the City Council in July.
At the March 20, 2017 Springfield City Council meeting, the Council voted 7-1 (Kristi Fulnecky was the no vote) to adopt Council Bill No. 2017-069 which is a Resolution authorizing the Mayor to sign a letter of support for a grant application by the St. Louis County Health Department to the Department of Justice for consortium funding for a PDMP. Yes, your tax money is being used to fund this, but only for two years. Then the city will be on the hook for at least $16,000 per year. They still must pass an ordinance to put it into effect. Stay tuned.
The second attachment is my slide presentation with notes. PDMP Presentation


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Shigemori, S: NIHON KADO BIJUTSU ZENSHU. (The Art of Japanese Flower Arranging). Kyoto, 1929. 28; 27; 27; 27; 27 folded leaves. 125 full page illustrations, the majority b/w. 5 vols. 31x23 cm. Stitched.
A pleasing five volume work from the early Showa period discussing the art of Japanese flower arranging. Each volume contains 25 full page illustrations of a flower arrangement accompanied by a page of descriptive text. The fine illustrations are a combination of black-and-white line drawings, b/w collotype and b/w photographic illustrations. A minority of the illustrations are lovely colour collotypes. All text in Japanese. Scarce.
Subjects: Botany Flower Arrangement
Available, as of: 12/02/2022
Was item 587 in printed List 202.
Record produced by Hanshan Tang Books, www.hanshan.com.

domain: literature


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This is by far some of the best night time photos of our landscape lighting. We lit this home in Leawood recently and captured the full moon rising over the roof. We took some with the house lights on and some with them off.
Spectacular. Let me know which pic you like the best. (You can click the images to see a larger version.)


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The NASCAR Xfinity Series returns to action this Saturday for the Boyd Gaming 300 at Las Vegas Motor Speedway. All 38 cars entered will race, barring any entry list changes.
Continuing his quest for 200 combined wins in NASCAR, Kyle Busch will pull triple duty and drive the Joe Gibbs Racing Extreme Concepts No. 18, while Austin Dillon will pull double duty and drive Kaulig Racing’s No. 10 Chevy.
The highly anticipated debut of Zane Smith comes to life as he’ll pilot the JR Motorsports No. 8, making him the third different driver for the No. 8 already this season. Junior Avila and Donald Theetge will also make their season debuts, driving for BJ Mcleod Motorsports (No. 99) and DGM Racing (No. 90), respectively.
The NASCAR Xfinity Series Boyd Gaming 300 at Las Vegas Motor Speedway is March 2 at 4 p.m. ET on Fox Sports 1.


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The beauty and perfection in spun glass collectible figurines will intrigue and amaze those of all ages. Its superb blown glass has delicate threading and chiseled detail. Spun glass is also known as lace glass or lace crystal. You will discover Spun glass figurines in a wide range of flowers, animals, boats, angels, people, carousels and much more. The only limitation to this art is the imagination of the artist.
As one of many new wave of glass blowers, René Steinke is pushing the boundaries of glass art and creating thrilling new work while teaching glass blowing classes to the community. As well as, Sac Artwork Glass is gathering place for artists – flame staff, metal artists, sculptors and painters who participate in Second Saturday occasions at the Sacramento studio.
There are even some that sell their shards in 1, 2 and 5 lb bags which have been tumbled in order that there are no sharp slicing edges remaining. What I like about these is that the pieces are already all mosaic sized and you simply use them as they’re. They make nice projects and their appearance is random and delightful! I order a number of pounds once in a while and when they arrive, I type them out by colour and put them in my jars ready to use for when I want them.
Alas, the primary of those home windows is misplaced to historical past, and their origins can solely be debated. Mosaics and jewelry makers are credited with creating the technique that created coloured glass artwork as we know it, which arose during a rise in church development. Nevertheless, coloured glass had been produced for the reason that days of the Egyptian and Roman empires, both of which surpassed others when it came to manufacturing small, stained glass objects.
After he misplaced the imaginative and prescient in his left eye as a result of a tragic 1976 auto accident, Dale relinquished his duties as chief glassblower at his studio. As a result of lack of depth notion he might now not perform the art of glass blowing. He started to direct different glassblowers who created artwork glass sculptures in line with his precise instructions. This was the start of his groundbreaking and revolutionary staff primarily based strategy to glass making.

domain: hobby


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ALSO AVAILABLE IN BEEF & FISH
HiLife Tempt Me! Petit portions
Chicken in Sauce 6 x 50g DISCONTINUED
Suggested Retail Price: £2.29 (per pack)
HiLife Tempt Me! Petit portions has been developed to give your cat a nutritious dinner with meaty chunks in a delicious herb-infused sauce. So delicious, they'll be purring for more! Free from artificial colours, preservatives, & wheat-gluten.
This is a complete food for adult cats.
If fed exclusively on HiLife tempt me! Petit portions, an average cat (4kg) should eat 3 - 4 pouches per day in at least two meals. If other products are fed during the day, remember to monitor total food intake and reduce the number of pouches fed. Always ensure your cat has access to plenty of fresh drinking water. Although every precatution has been take in the preparation of this recipe, it may contain small bones.
Chicken with Tarragon Sauce - Chicken (40%, incl.chicken breast 14%), soy protein, potato starch, sunflower oil, minerals, tarragon (0.2%), sugar, taurine
Chicken with Rosemary Sauce - Chicken (40%, incl.chicken breast 14%), soy protein, potato starch, sunflower oil, minerals, rosemary (0.2%), sugar, taurine
Nutritional Additives - Vitamin A 10,350 iu/kg, Vitamin D3 1,050 iu/kg, Ferrous sulphate monohydrate 51.9 mg/kg, Potassium iodide 10.4 mg/kg, Copper sulphate pentahydrate 4.1 mg/.kg, Manganese sulphate monohydrate 6.2 mg/kg, Zinc sulphate heptahydrate 207.4 mg/kg, Natural Colourants.
Protein 10.5%, Fat Content 7.5%, Crude Fibres 0.5%, Inorganic Matter 1.5%, Moisture 78.0%
Lindsay Mason - May 2017
Bought these, along with the beef and fish versions, yesterday for my cat who has been off her food. She's been to the vet and treated with antibiotics but getting her to eat was proving tricky - even her favourite fresh chicken was turned down! I was very excited to see these in the store as I thought they may just whet her appetite. Boy, was I right! She is absolutely loving them! She has just wolfed down the chicken in tarragon and has really enjoyed each one 've given her so far. I like the 50 g pouches too as I can give her enough to eat in one go without having to put some in the fridge or waste food. Brilliant product and I highly recommend them - just hope I can continue to source them (no pun intended!) as they are not a regular option at my local store.

domain: pet


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Chapter six was very informative, it showed how words and pictures really come together to create this great piece of work we call comics. I liked how it explained the evolution of both. How they have both changed over the years. And how they have come together. In this chapter i got a better understanding of how the words and the images of comics work together. Words and pictures simply add to one another. They enhance each other and ensure that the meaning will be clear to the reader. The use of words and images really does make comics limitless. Comics can do just about anything the options are endless when it comes to comics.
10 February 2011
Hurray for Show and Tell! The Balance of Pictures and Words.
Chapter six in the book Understanding Comics is all about "show and tell". Show and tell you may ask? Isn't that something that kindergartners do? Well no, Scott McCloud explains in chapter six how comics are all about show and tell. But first he explains the history of both words and pictures. When he is talking about the history he shows how they have both evolved. After explaining the history he goes into how they have come together to create comics. The pictures show whats going on and the words explain in depth whats happening. There are different types of show and tell when it comes to comics. The different ones are word specific-which have pictures to show but need text to complete it, picture specific-words do more they are like a "soundtrack", duo specific-words and pictures do the same thing, additive- words or pictures "amplify" or "elaborate", parallel- the words and pictures follow different paths, montage- the words are literally apart of the picture, interdependent- the words and pictures go "hand in hand" they both need each other. Pictures and words work together when it comes to comics. They are a team and have to work together to accomplish a goal. There needs to be balance when it comes to comics. Words and pictures both can't lead. Pictures usually take the lead when it comes to comics, but the words can be used to explore or go more into depth and expand on an idea.


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4rabet India website is a legal system for internet gambling. However, right now, this element is virtually not jeopardized since a lot of the systems have discovered to conserve the individual information as well as funds of gamers in a dependable means. To play on the internet casino video games with genuine cash, gamers need to select among the widely known gambling systems, register there, make a down payment as well as begin to play beneficially on their own. Free rotates no down payment mobile casino in the U.S.A. would certainly be the finest technique to raise your revenue together with satisfaction also. An online casino can play a range of casino video games via numerous specialized websites or mobile applications. Exactly How to Play Casino Gamings Online with Genuine Cash?
Exactly how to take out cash from an internet casino? On the internet, casino websites supply a variety of withdrawal approaches comparable to down payment alternatives. 4rabet online casino approves credit/debit cards as well as on the internet repayment choices e-wallets such as Visa, Mastercard, Skrill, Neteller, Paytm as well as PhonePe. Is 4rabet lawful in India? Which is the ideal online casino in India? SattaMatka video game was presented by a male called Ratan Khatri as well as this video game made a massive name in India. Invite to real Las Las vega on the internet casino port video game as well as rotate the finest timeless 777 ports casino equipment The online ports with the most effective as well as greatest payments are those that, such as the Huge Moolah collection from software program programmer Microgaming. The order will certainly be verified by the solution reps after the information has readout.
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