CCK-induced pancreatic growth is not limited by mitogenic capacity in mice

In mice fed trypsin inhibitor (camostat) to elevate endogenous CCK, pancreatic growth plateaus by 7 days. It is unknown whether this represents the maximum growth capacity of the pancreas. To test the ability of CCK to drive further growth, mice were fed chow containing camostat (0.1%) for 1 wk, then fed standard chow for 1 wk, and finally returned to the camostat diet for a week. Pancreatic mass increased to 245% of initial value (iv) following 1 wk of camostat feeding, decreased to 147% iv following a 1 wk return to normal chow, and increased to 257% iv with subsequent camostat feeding. Camostat feeding was associated with significant increases in circulating CCK and changes in pancreatic mass were paralleled by changes in protein and DNA content. Moreover, regression of the pancreas following camostat feeding was associated with changes in the expression of the autophagosome marker LC3. Pancreatic protein synthetic rates were 130% of control after 2 days on camostat but were equivalent to control after 7 days. Changes in the phosphorylation of 4E-BP1 and S6, downstream effectors of mammalian target of rapamycin (mTOR), paralleled changes in protein synthetic rates. Cellular content of Akt, an upstream activating kinase of mTOR, decreased after 7 days of camostat feeding whereas expression of the E3 ubiquitin-ligases and the cell cycle inhibitor p21 increased after 2 days. These results indicate that CCK-stimulated growth of the pancreas is not limited by acinar cell mitogenic capacity but is due, at least in part, to inhibition of promitogenic Akt signaling.     

Dry matter accumulation in citrus fruit is not limited by transport capacity of the pedicel

The vascularization of the pedicel in Marisol clementine (Citrus clementina Hort. ex Tanaka) has been characterized in relation to fruit growth. Phloem and xylem formation occurred during the first half of the period of fruit growth. Phloem cross-sectional area reached its maximum value by the end of fruitlet abscission, 78 d after anthesis (DAA), shortly after the rate of accumulation of dry matter in fruitlets reached its maximum value. Secondary xylem formation occurred until day 93, well after the end of fruitlet abscission. At fruit maturity, xylem accounted for 42-46 % of the cross-section of the pedicel. Vessels differentiated in this late-formed xylem. Formation of phloem and early xylem was directly related to fruitlet size (and growth rate). Differences in the rate of formation of conductive tissues in the pedicel of the developing fruitlets followed rather than preceded the differences in growth rate. Specific mass transfer (SMT) in the phloem was highest in the fastest growing fruitlets, and peaked during the late stages of fruitlet abscission (72-78 DAA) and during the main period of fruit growth (107-121 DAA). Application of a synthetic auxin to developing fruits, either at the end of flowering (2,4-D) or by day 64 after flowering (2,4-DP), increased the growth rate of the fruit and fruit size at maturity (8-13 % increase in fruit diameter at maturity). These auxin applications also enhanced the formation of conductive tissues in the pedicel, with a specific effect on phloem formation. Applying auxin at flowering resulted in a reduction in the phloem SMT by days 72-78, whereas auxin application on day 64 increased this parameter. Despite this difference in behaviour, which resulted from the different time-course of the growth response of the fruit to auxin applications, these applications increased fruit size to a similar extent. Severing 37 % of the phloem of the pedicel during the main period of fruit growth resulted in an increase in the specific mass transfer in the phloem but had no influence on fruit growth. These observations demonstrate that the transport capacity in the phloem of the pedicel does not limit fruit growth and, within the limits of our experiments, an increase in demand by the fruit appeared to be matched by an increase in SMT. The dependence of late xylem formation (after the period of fruitlet abscission) on fruitlet growth was demonstrated in Salustiana orange [Citrus sinensis (L.) Osbeck] by means of controlling fruit growth through the manipulation of leaf area. Fruit growth at this time was more closely related to leaf area than to carbohydrate levels, suggesting that it may be limited by current photosynthesis.     

Growth of Saccharomyces cerevisiae is controlled by its limited respiratory capacity: Formulation and verification of a hypothesis

A novel mechanistic model for the growth of baker's yeast on glucoseis presented. It is based on the fact that glucose degradation proceeds via two pathways under conditions of aerobic ethanol formation. Part is metabolized oxidatively and part reductively, with ethanol being the end product of reductive energy metabolism. The corresponding metabolic state is designated oxidoreductive. Ethanol can be used oxidatively only. Maximum rates of oxidative glucose and ethanol degradation are governed by the respiratory capacity of the cells. The model is formulated by using the stoichiometric growth equations for pure oxidative and reductive (fermentative) glucose and ethanol metabolism. Together with the experimentally determinable yield coefficients (Y(X/S)) for the respective metabolic pathways, the resulting equation system is sufficiently determined. The superiority of the presented model over hitherto published ones is based on two essential novelities. (1) The model was developed on experimentally easily accessible parameters only. (2) For the modeling of aerobic ethanol formation, the substrate flow was split into two simultaneously operating (i.e., in parallel) metabolic pathways that exhibit different but constant energy-generating efficiencies (respiration and fermentation) and consequently different and constant biomass yields (Y(X/S)). The model allows the prediction of experimental data without parameter adaption in a biologically dubious manner.     

Parent-offspring conflict that is not limited by degree of kinship

Models are presented showing that, under certain conditions, when multiple offspring are reared simultaneously, the magnitude of offspring selfishness is independent of offspring-offspring genetic relatedness. When the cost (θ) of the selfish offspring's behavior approaches zero, alleles causing offspring selfishness spread as long as $\text{c}\text{b}$ is positive.In general, simultaneous offspring are predicted to behave more selfishly than sequential offspring.Stable equilibria between alleles for selfish and non-selfish behavior are predicted for certain selfish alleles.     

Egg-laying capacity is limited by carotenoid pigment availability in wild gulls Larus fuscus

In birds, experimentally increased egg production can reduce maternal condition, parenting ability and survival, and the quality of the eggs themselves. Such costs probably reflect resource limitation, but the identity of the resource(s) in question remains unclear. Carotenoids are antioxidants and immunomodulants that birds can only obtain in their diet. Trade-offs in the allocation of limiting carotenoids between somatic maintenance and egg production could therefore be an important factor underlying reproductive costs. We show that in wild lesser black-backed gulls, Larus fuscus, dietary carotenoid availability (i) constrained the capacity to re-lay following clutch removal; and (ii) affected the relationship between yolk mass and egg mass. However, whether carotenoids are limiting for egg production directly, by stimulating the synthesis or antioxidant protection of yolk precursors, or indirectly via effects on maternal health, requires further study.     

Respiration rate in maize roots is related to concentration of reduced nitrogen and proliferation of lateral roots

The relationship between specific rate of respiration (respiration rate per unit root dry weight) and concentration of reduced nitrogen was examined for maize ( Zea mays L.) roots. Plants with 2 primary nodal root axes were grown for 8 days in a split-root hydroponic system in which NO-₃ was supplied to both axes at 1.0 mol m⁻³, to one axis at 1.0 mol m⁻³ and the other axis at 0.0 mol m⁻³ or to both axes at 0.0 mol m⁻³ Respiration rates and root characteristics were measured at 2-day intervals. Specific rate of respiration was positively correlated in a nonlinear relationship with concentration of reduced nitrogen. The lowest specific rates of respiration occurred when neither axis received exogenous NO⁻³ and the concentration of reduced nitrogen in the axes was less than 9 mg g⁻¹. The greatest rates occurred in axes that were actively absorbing NO⁻³ and contained more than 35 mg g⁻¹ of reduced nitrogen. At 23 mg g⁻¹ of reduced nitrogen, below which initiation of lateral branches was decreased by 30–50%. specific rate of respiration was 17% greater for roots actively absorbing NO⁻³ than for roots not absorbing NO⁻³ Increases in specific rate of respiration associated with concentrations of reduced nitrogen greater than 23 mg g⁻¹ were concluded to be attributable primarily to proliferation of lateral branches.     

Inherent capacity for lipogenesis or dietary fat retention is not increased in obesity-prone rats

Obesity results from positive energy balance and, perhaps, abnormalities in lipid and glycogen metabolism. The purpose of this study was to determine whether differences in lipogenesis, retention of dietary fat, and/or glycogenesis influenced susceptibility to dietary obesity. After 1 wk of free access to a high-fat diet (HFD; 45% fat by energy) rats were separated on the basis of 1 wk body weight gain into obesity-prone (OP; > or =48 g) or obesity-resistant groups (OR; < or =40 g). Rats were either studied at this time (OR1, OP1) or continued on the HFD for an additional 4 wk (OR5, OP5). Weight gain and energy intake were greater (P < or = 0.05) in OP vs. OR at both 1 (53 +/- 2 vs. 34 +/- 1 g; 892 +/- 27 vs. 755 +/- 14 kcal) and 5 (208 +/- 7 vs. 170 +/- 7 g; 4,484 +/- 82 vs. 4,008 +/- 72 kcal) wk, respectively. Rats were injected with (3)H(2)O and were either provided free access to an HFD meal containing labeled fatty acids (fed; n = 10 or 11/group) or were fasted (n = 10/group) overnight. The amount of food or (14)C tracer eaten overnight was equivalent between OP and OR rats. In liver, the fraction of (3)H retained in glycogen or lipid was not significantly different between OR and OP groups. Retention of dietary fat in the liver was not increased in OP rats. In adipose tissue, retention of (3)H was approximately 49% greater (P < or = 0.05) in OP1 vs. OR1 and approximately 30% greater in OP5 vs. OR5, but retention of dietary fat was not elevated in OP vs. OR. At the same time, fat pad weight (sum of epididymal, retroperitoneal, mesenteric) was 49% greater in OP1 rats vs. OR1 rats and 65% greater in OP5 vs. OR5 rats (P < or = 0.05). Thus a greater capacity for lipogenesis or retention of dietary fat does not appear to be included in the OP phenotype. The characteristic increase in energy intake associated with OP rats appears to be necessary and critical to accelerated weight and fat gain.     

Gravitropic curvature of maize roots is not preceded by rootcap asymmetry

We tested whether the first response to gravistimulation is an asymmetry in the root tip that results from differential growth of the rootcap itself. The displacement of markers on the rootcap surface of maize (Zea mays L. cv. Merit) roots was quantified from videotaped images using customized software. The method was sensitive enough to detect marker displacements down to 15 microns and root curvature as early as 8 min after gravistimulation. No differential growth of the upper and lower sides of the cap occurred before or during root curvature. Fewer than a third of all gravistimulated roots developed an asymmetrical outline of the root tip after curvature had started, and this asymmetry did not occur in the rootcap itself. Our data support the view that the regions of gravitropic sensing and curvature are spatially separate during all phases of gravitropism in maize roots.     

Red pigment-concentrating hormone is not limited to crustaceans

A peptide that was previously assumed to occur exclusively in crustaceans is found in the corpora cardiaca of the stinkbug, Nezara viridula. The sequence of the peptide was deduced from the multiple MS(N) electrospray mass data as that of an octapeptide: pGlu-Ile/Leu-Asn-Phe-Ser-Pro-Gly-Trp amide. This peptide with Leu at position 2 is known as crustacean red pigment-concentrating hormone and code-named Panbo-RPCH. The ambiguity about the amino acid at position 2, Leu or Ile, was solved by isolating the peptide in a single-step by reversed-phase HPLC and establishing co-elution with authentic Panbo-RPCH but not with the Ile(2)-analog. When injected into stinkbugs, synthetic Panbo-RPCH elicited an increase of lipids in the haemolymph. Thus, it is assumed that Panbo-RPCH functions in the stinkbug as a lipid-mobilizing hormone.     

Miniaturized orb-weaving spiders: behavioural precision is not limited by small size

The special problems confronted by very small animals in nervous system design that may impose limitations on their behaviour and evolution are reviewed. Previous attempts to test for such behavioural limitations have suffered from lack of detail in behavioural observations of tiny species and unsatisfactory measurements of their behavioural capacities. This study presents partial solutions to both problems. The orb-web construction behaviour of spiders provided data on the comparative behavioural capabilities of tiny animals in heretofore unparalleled detail; species ranged about five orders of magnitude in weight, from approximately 50-100mg down to some of the smallest spiders known (less than 0.005mg), whose small size is a derived trait. Previous attempts to quantify the 'complexity' of behaviour were abandoned in favour of using comparisons of behavioural imprecision in performing the same task. The prediction of the size limitation hypothesis that very small spiders would have a reduced ability to repeat one particular behaviour pattern precisely was not confirmed. The anatomical and physiological mechanisms by which these tiny animals achieve this precision and the possibility that they are more limited in the performance of higher-order behaviour patterns await further investigation.     

Determinate growth in a paternal mouthbrooding fish whose reproductive success is limited by buccal capacity

The life history of the paternal mouthbrooding cardinal fish Apogon doederleini was investigated in the temperate waters of Japan, with particular reference to its growth and reproductive rate. Both males and females almost ceased to grow at age 3 years, although living to 7 years of age. Their growth pattern, represented by the relative size at sexual maturity to the asymptotic size and the von Bertalanffy growth coefficient, was among the most determinate in ectothermic vertebrates. Brood size just before hatching increased in proportion to the second power of the body size of the brooding male, and correlated more positively with the male's than the female's body size, suggesting that it was limited by the male's buccal capacity. The estimated total number of broods hatched in a breeding season showed a weak or no correlation with the body size or age in either sex. Using life-history parameters based on data of A. doederleini, a simulation model of energy allocation without considering sexual interaction revealed that the optimal growth pattern shows an indeterminate growth that differs greatly from the actual growth pattern of A. doederleini. This suggests that there are some brooding constraints to size-advantage of reproductive success in this species. The possible mechanism of such reproductive constraint is discussed.     

Nitric oxide uptake by erythrocytes is primarily limited by extracellular diffusion not membrane resistance

The process of NO transfer into erythrocytes (RBCs) is of critical biological importance because it regulates the bioavailability and diffusional distance of endothelial-derived NO. It has been reported that the rate of NO reaction with oxyhemoglobin (Hb) within RBCs is nearly three orders of magnitude slower than that by equal amounts of free oxyhemoglobin. Consistent with early studies on oxygen uptake by RBCs, the process of extracellular diffusion was reported to explain this much lower NO uptake by RBC encapsulated Hb (Liu, X., Miller, M. J., Joshi, M. S., Sadowska-Krowicka, H., Clark, D. A., and Lancaster, J. R., Jr. (1998) J. Biol. Chem. 273, 18709-18713). However, it was subsequently proposed that the RBC membrane provides the main resistance to NO uptake rather than the process of extracellular diffusion (Vaughn, M. W., Huang, K. T., Kuo, L., and Liao, J. C. (2000) J. Biol. Chem. 275, 2342-2348). This conclusion was based on competition experiments that were assumed to be able to determine the rate constant of NO uptake by RBCs without extracellular diffusion limitation. To test the validity of this hypothesis, we theoretically analyzed competition experiments. Here, we show that competition experiments do not eliminate the extracellular diffusion limitation. Simulation of the competition data indicates that the main resistance to NO uptake by RBCs is caused by extracellular diffusion in the unstirred layer surrounding each RBC but not by the RBC membrane. This extracellular diffusion resistance is responsible for preventing interference of NO signaling in the endothelium without the need for special NO uptake by intracellular hemoglobin or a unique membrane resistance mechanism.     

By-product formation during exposure of respiring Saccharomyces cerevisiae cultures to excess glucose is not caused by a limited capacity of pyruvate carboxylase

Upon exposure to excess glucose, respiring cultures of Saccharomyces cerevisiae produce substantial amounts of ethanol and acetate. A possible role of a limited anaplerotic capacity in this process was investigated by overexpressing pyruvate carboxylase and by replacing it with a heterologous enzyme (Escherichia coli phosphoenolpyruvate carboxylase). Compared to the wild-type, neither the pyruvate carboxylase (Pyc)-overexpressing nor the transgenic strain exhibited reduced by-product formation after glucose pulses to aerobic glucose-limited chemostat cultures. An increased intracellular malate concentration was observed in the two engineered strains. It is concluded that by-product formation in S. cerevisiae is not caused by a limited anaplerotic capacity.     

Maximal oxygen uptake is not limited by a central nervous system governor.

We tested the hypothesis that the work of the heart was not a limiting factor in the attainment of maximal oxygen uptake (VO2 max). We measured cardiac output (Q) and blood pressures (BP) during exercise at two different rates of maximal work to estimate the work of the heart through calculation of the rate-pressure product, as a part of the ongoing discussion regarding factors limiting VO2 max. Eight well-trained men (age 24.4 +/- 2.8 yr, weight 81.3 +/- 7.8 kg, and VO2 max 59.1 +/- 2.0 ml x min(-1) x kg(-1)) performed two maximal combined arm and leg exercises, differing 10% in watts, with average duration of time to exhaustion of 4 min 50 s and 3 min 40 s, respectively. There were no differences between work rates in measured VO2 max, maximal Q, and peak heart rate between work rates (0.02 l/min, 0.3 l/min, and 0.8 beats/min, respectively), but the systolic, diastolic, and calculated mean BP were significantly higher (19, 5, and 10 mmHg, respectively) in the higher than in the lower maximal work rate. The products of heart rate times systolic or mean BP and Q times systolic or mean BP were significantly higher (3,715, 1,780, 569, and 1,780, respectively) during the higher than the lower work rate. Differences in these four products indicate a higher mechanical work of the heart on higher than lower maximal work rate. Therefore, this study does not support the theory, which states that the work of the heart, and consequently VO2 max, during maximal exercise is hindered by a command from the central nervous system aiming at protecting the heart from being ischemic.     

Prestin-driven cochlear amplification is not limited by the outer hair cell membrane time constant

Outer hair cells (OHCs) provide amplification in the mammalian cochlea using somatic force generation underpinned by voltage-dependent conformational changes of the motor protein prestin. However, prestin must be gated by changes in membrane potential on a cycle-by-cycle basis and the periodic component of the receptor potential may be greatly attenuated by low-pass filtering due to the OHC time constant (τ(m)), questioning the functional relevance of this mechanism. Here, we measured τ(m) from OHCs with a range of characteristic frequencies (CF) and found that, at physiological endolymphatic calcium concentrations, approximately half of the mechanotransducer (MT) channels are opened at rest, depolarizing the membrane potential to near -40 mV. The depolarized resting potential activates a voltage-dependent K+ conductance, thus minimizing τ(m) and expanding the membrane filter so there is little receptor potential attenuation at the cell's CF. These data suggest that minimal τ(m) filtering in vivo ensures optimal activation of prestin.     

Respiration capacity and consequences in Lactococcus lactis

We recently reported that the well-studied fermenting bacterium Lactococcus lactis could grow via a respirative metabolism in the presence of oxygen when a heme source is present. Respiration induces profound changes in L. lactis metabolism, and improvement of oxygen tolerance and long-term survival. Compared to usual fermentation conditions, biomass is approximately doubled by the end of growth, acid production is reduced, and large amounts of normally minor end products accumulate. Lactococci grown via respiration survive markedly better after long-term storage than fermenting cells. We suggest that growth and survival of lactococci are optimal under respiration-permissive conditions, and not under fermentation conditions as previously supposed.Our results reveal the uniqueness of the L. lactis respiration model. The well-studied aerobic bacteria express multiple terminal cytochrome oxidases, which assure respiration all throughout growth; they also synthesize their own heme. In contrast, the L. lactis cydABgenes encode a single cytochrome oxidase (bd), and heme must be provided. Furthermore, cydAB genes mediate respiration only late in growth. Thus, lactococci exit the lag phase via fermentation even if heme is present, and start respiration in late exponential phase. Our results suggest that the spectacularly improved survival is in part due to reduced intracellular oxidation during respiration. We predict that lactococcal relatives like the Enterococci, and some Lactobacilli, which have reported respiration potential, will display improved survival under respiration-permissive conditions.     

The up-regulation of utrophin is not limited to muscular dystrophies

Immunohistochemical reactivity for utrophin has been recorded in 45 biopsies from patients with various neuromuscular diseases. The upregulation of utrophin on the extrajunctional sarcolemma has been found in dystrophinopathies, other muscular dystrophies, congenital myopathies, inflammatory myopathies, neurogenic muscle disorders (diabetic neuropathy, amyotrophic lateral sclerosis and spinal muscular atrophies), minimal change myopathies as well as in some normal biopsies.