Is the Rate of Metabolic Ageing and Survival Determined by Basal Metabolic Rate in the Zebra Finch?

The relationship between energy metabolism and ageing is of great interest because aerobic metabolism is the primary source of reactive oxygen species which is believed to be of major importance in the ageing process. We conducted a longitudinal study on captive zebra finches where we tested the effect of age on basal metabolic rate (BMR), as well as the effect of BMR on the rate of metabolic ageing (decline in BMR with age) and survival. Basal metabolic rate declined with age in both sexes after controlling for the effect of body mass, indicating a loss of functionality with age. This loss of functionality could be due to accumulated oxidative damage, believed to increase with increasing metabolic rate, c.f. the free radical theory of ageing. If so, we would expect the rate of metabolic ageing to increase and survival to decrease with increasing BMR. However, we found no effect of BMR on the rate of metabolic ageing. Furthermore, survival was not affected by BMR in the males. In female zebra finches there was a tendency for survival to decrease with increasing BMR, but the effect did not reach significance (P<0.1). Thus, the effect of BMR on the rate of functional deterioration with age, if any, was not strong enough to influence neither the rate of metabolic ageing nor survival in the zebra finches.     

Wolf Movement Patterns: a Key to Estimation of Kill Rate?

Abstract: To estimate wolf (Canis lupus) kill rates from fine-scale movement patterns, we followed adult wolves in 3 territories of the Scandinavian wolf population using Global Positioning Systems (GPS) during the winters of 2001–2003. The resulting 6 datasets of 62–84 study days gave a total of 8,747 hourly GPS positions. We visited clusters of positions in the field on average 8.8 days after positioning and found moose (Alces alces) killed by wolves during the study period on 74 (8%) of the 953 clusters. The number of positions and visits to a cluster, their interaction, and the proportion of afternoon positions were significant fixed effects in mixed logistic-regression models predicting the probability of a cluster containing a wolf-killed moose. The models, however, displayed a poor goodness-of-fit and were not a suitable tool for estimating kill rates from positioning data alone. They might be used to reduce fieldwork by excluding unlikely clusters, although the reduction was not substantial. We discuss proximate factors (i.e., human disturbance and access to prey) as well as ultimate factors (i.e., social organization, intra-guild dominance, and litter size) as potential causes of the observed high temporal and spatial variation in prey-handling. For similar future kill-rate studies, we recommend increasing field efforts and shortening positioning intervals.     

Is the Mutation Rate Lower in Genomic Regions of Stronger Selective Constraints?

A study of the plant Arabidopsis thaliana detected lower mutation rates in genomic regions where mutations are more likely to be deleterious, challenging the principle that mutagenesis is blind to its consequence. To examine the generality of this finding, we analyze large mutational data from baker''s yeast and humans. The yeast data do not exhibit this trend, whereas the human data show an opposite trend that disappears upon the control of potential confounders. We find that the Arabidopsis study identified substantially more mutations than reported in the original data-generating studies and expected from Arabidopsis'' mutation rate. These extra mutations are enriched in polynucleotide tracts and have relatively low sequencing qualities so are likely sequencing errors. Furthermore, the polynucleotide “mutations” can produce the purported mutational trend in Arabidopsis. Together, our results do not support lower mutagenesis of genomic regions of stronger selective constraints in the plant, fungal, and animal models examined.     

Is Aberrant DNA Methylation a Key Factor in Molar Incisor Hypomineralization?

Molar incisor hypomineralization (MIH) is a qualitative disturbance of the enamel of the permanent molars and/or incisors. Its etiology is not clearly defined but is connected with different factors occurring before and after birth. It remains difficult to identify a single factor or group of factors, and the problem is further complicated by various overlapping mechanisms. In this study, we attempted to determine whether DNA methylation—an epigenetic mechanism—plays a key role in the etiology of MIH. We collected the epithelium of the oral mucosa from children with MIH and healthy individuals and analyzed its global DNA methylation level in each child using a 5-mC DNA ELISA kit after DNA isolation. There was no statistically significant difference between the global DNA methylation levels in the study and control groups. Then, we also analyzed the associations of the DNA methylation levels with different prenatal, perinatal, and postnatal factors, using appropriate statistical methods. Factors such as number of pregnancies, number of births, type of delivery, varicella infection (under 3 years old), and high fever (under 3 years old) were significantly important. This work can be seen as the first step towards further studies of the epigenetic background of the MIH etiology.     

Is progesterone a pre-hormone in the CNS?

In this paper, experimental evidences have been presented indicating that progesterone per se appears to be a powerful modulatory steroid of presynaptic striatal dopaminergic terminals of the central nervous system of the rat. This effect of the progesterone signal is concentration as well as infusion mode dependent. Low pulsatile doses of the steroid positively modulate the mechanism by which dopamine terminals respond to amphetamine stimulation and increase tissue dopamine concentration. Whereas, continuous and/or high doses of this steroid negatively modulate the response of the dopamine terminals to amphetamine stimulation and decreases tissue dopamine concentration. This effects occurs through a membrane mediated mechanism either upon the dopamine neuron directly and/or upon an interneuron. Pregnanolone a 5- beta-3 beta-metabolite of progesterone known to activate the hypothalamic LHRH neural apparatus at the level of the hypothalamus of ovariectomized estrogen primed rats in both in vitro as well as in vivo preparations was completely ineffective at the level of the corpus striatum of similar animal preparations. Therefore, it is reasonable to assume that site specific mechanisms exist within the central nervous system which may control differentially the final action of progesterone. In the hypothalamus, pregnanolone appears to be the final signal for its action on the LHRH neural apparatus, whereas in the corpus striatum, the steroid per se, and dependent on the modality and/or the strength of the signal can either directly or indirectly up-regulate (stimulatory component) or down-regulate (inhibitory component) the activity of striatal dopaminergic terminals.     

E2P State Stabilization by the N-terminal Tail of the H,K-ATPase ��-Subunit Is Critical for Efficient Proton Pumping under in Vivo Conditions

The catalytic α-subunits of Na,K- and H,K-ATPase require an accessory β-subunit for proper folding, maturation, and plasma membrane delivery but also for cation transport. To investigate the functional significance of the β-N terminus of the gastric H,K-ATPase in vivo, several N-terminally truncated β-variants were expressed in Xenopus oocytes, together with the S806C α-subunit variant. Upon labeling with the reporter fluorophore tetramethylrho da mine-6-maleimide, this construct can be used to determine the voltage-dependent distribution between E₁P/E₂P states. Whereas the E₁P/E₂P conformational equilibrium was unaffected for the shorter N-terminal deletions βΔ4 and βΔ8, we observed significant shifts toward E₁P for the two larger deletions βΔ13 and βΔ29. Moreover, the reduced ΔF/F ratios of βΔ13 and βΔ29 indicated an increased reverse reaction via E₂P → E₁P + ADP → E₁ + ATP, because cell surface expression was completely unaffected. This interpretation is supported by the reduced sensitivity of the mutants toward the E₂P-specific inhibitor {"type":"entrez-protein","attrs":{"text":"SCH28080","term_id":"1053015931","term_text":"SCH28080"}}SCH28080, which becomes especially apparent at high concentrations (100 μm). Despite unaltered apparent Rb⁺ affinities, the maximal Rb⁺ uptake of these mutants was also significantly lowered. Considering the two putative interaction sites between the β-N terminus and α-subunit revealed by the recent cryo-EM structure, the N-terminal tail of the H,K-ATPase β-subunit may stabilize the pump in the E₂P conformation, thereby increasing the efficiency of proton release against the million-fold proton gradient of the stomach lumen. Finally, we demonstrate that a similar truncation of the β-N terminus of the closely related Na,K-ATPase does not affect the E₁P/E₂P distribution or pump activity, indicating that the E₂P-stabilizing effect by the β-N terminus is apparently a unique property of the H,K-ATPase.The gastric H,K-ATPase fulfills the remarkable task of pumping protons against a more than 10⁶-fold concentration gradient. H⁺ extrusion is coupled to countertransport of an equal number of K⁺ ions for each ATP molecule hydrolyzed, resulting in an electroneutral overall process (1). Characteristic for all P-type ATPases, the enzyme cycles between the two principal conformational states (E₁ and E₂) and the corresponding phosphointermediates (E₁P and E₂P), which are formed by reversible phosphorylation of an aspartate residue in the highly conserved DKTGTLT motif. According to a Post-Albers-like reaction scheme (see Fig. 1A), the conformational E₁P → E₂P transition converts the high H⁺/low K⁺ affinity of the cation binding pocket into a low H⁺/high K⁺ affinity binding site, hence enabling proton release into the stomach lumen and subsequent binding of extracellular K⁺. Because the pump faces a lumenal proton concentration of ∼150 mm (2), proton release is probably the energetically most demanding step in the reaction cycle. Thus, during the conformational E₁P → E₂P transition, enormous pK_a changes of the H⁺-coordinating residues have to occur that most likely involve the rearrangement of a positively charged lysine side chain (Lys-791 in rat H,K-ATPase) (3).Open in a separate windowFIGURE 1.Post-Albers scheme (A) and cryo-EM structural representation of pig gastric H,K-ATPase in the fluoroaluminate-bound pseudo-E₂P state (B). A, Post-Albers scheme of the proposed reaction cycle of the gastric H,K-ATPase. E₁P/E₂P conformational states giving rise to voltage jump-induced fluorescence changes of TMRM-labeled H,K-ATPase molecules are highlighted (gray box). B, structural representation based on the cryo-EM structure of the pig gastric H,K-ATPase (surface or mesh, contoured at 1 σ; EM Data Bank code 5104) and the corresponding homology model (schematic; Protein Data Bank code 3IXZ). Inset, a close-up view (from the right side of the molecule) showing the putative interaction sites of the β-subunit N terminus with the P-domain (red arrow) and αTM3 (black arrow), respectively. Color coding is indicated in the figure.All P₂-type ATPases share a common catalytic α-subunit, composed of 10 transmembrane domains harboring the ion-binding sites and a large cytoplasmic loop with the nucleotide-binding domain, the phosphorylation domain (P-domain),² and the actuator domain (A-domain) (4). However, a unique feature of K⁺-transporting Na,K- and H,K-ATPase enzymes is the requirement for an accessory β-subunit, which is indispensable for proper folding, maturation, and plasma membrane delivery (5, 6). Despite only 20–30% overall sequence identity between the H,K-ATPase β-subunit and the Na,K β-isoforms, the topogenic structure is similar: a short N-terminal cytoplasmic tail, followed by a single transmembrane segment and a large extracellular C-terminal domain with glycosylation sites and disulfide-bridging cysteines. Numerous studies have demonstrated that the β-subunit of the Na,K-ATPase is more than just a chaperone for the α-subunit, being also required for proper ion transport activity of the holoenzyme. In fact, it has been discovered that different cell- and tissue-specific β-isoforms have distinct effects on the cation affinities (7–9). Furthermore, it was shown that mutational changes in all three topogenic domains of the Na,K-ATPase β-subunit (10–19) as well as chemical interference with disulfide-forming cysteines in the Na,K-ATPase β-subunit ectodomain (20–22) affect the cation transport properties of the sodium pump. Finally, conformational changes in the β-subunit during the Na,K-ATPase reaction cycle were demonstrated by proteolytic digestion studies (23) and voltage clamp fluorometry (24).Far less is known about the functional significance of the single H,K-ATPase β-isoform, especially about its potential impact on cation transport (reviewed in Refs. 25 and 26). We have proven recently that E₂P state-specific transmembrane interactions between residues in αTM7 and two highly conserved tyrosines in the βTM of both Na,K- and H,K-ATPase significantly stabilize the E₂P conformation (19). Mutational disruptions of this interaction resulted in substantial shifts toward E₁P and severely affected H⁺ secretion, which highlighted the physiological relevance of this E₂P state stabilization. Notably, according to the recently published cryo-EM structure of pig gastric H,K-ATPase in the pseudo-E₂P state (27), the N-terminal tail of the β-subunit makes direct contact with the phosphorylation domain of the α-subunit (see Fig. 1B), thus indicating an additional E₂P state stabilization mediated by the β-N terminus. Although this idea was further supported by biochemical studies on N-terminally truncated mutants, direct evidence for this putative E₂P-stabilizing interaction and its potential significance for ion transport in intact cells is still lacking.Here, we demonstrate for the first time the functional importance of the gastric H,K-ATPase β-subunit N terminus in living cells under in vivo conditions: voltage clamp fluorometry, Rb⁺ flux, and {"type":"entrez-protein","attrs":{"text":"SCH28080","term_id":"1053015931","term_text":"SCH28080"}}SCH28080 sensitivity measurements revealed E₁P-shifted, ion transport-impaired phenotypes for two N-terminally truncated H,K β-variants, thus substantiating the E₂P-stabilizing effect of the β-N terminus suggested by the recent cryo-EM structure.     

Is the originality of a species measurable?

In this paper, we introduce the concept of 'originality of a species within a set' in order to indicate the average rarity of all the features belonging to this species. Using a phylogenetic tree of 70 species of New World terrestrial Carnivora, we suggest measuring the originality by a probability distribution. This maximizes the expected number of features shared by two species randomly drawn from the set. By using this new index, we take account of branch lengths whereas current indices of originality focus on tree topology. As a supplement to Nee and May's optimizing algorithm, we find that originality must be one of the criteria used in conservation planning.     

Is virtual reality a useful tool in the teaching of physiology?

This opinion statement points out some of the considerations and pitfalls in using virtual reality computer programs in the teaching of life sciences. Emphasis is placed on the possibility of such programs leading to reductionist thinking including how reductionist thinking could foster the formation of misconceptions. Negative feedback is used as the classic example of reductionist thinking in physiological regulation, including how classic negative feedback is inconsistent with evidence of complexity in living systems. This statement concludes that virtual reality can be a useful tool in the teaching of physiology so long as the complexity of living systems is taken into account.     

Is beauty in the eye of the beholder?

Theories of beauty were evaluated by requiring subjects to “evolve” a beautiful female face using a Genetic Algorithm. In this procedure, a computer program generated a small population of faces (first generation of phenotypes) from a set of random binary strings (genotypes). Genotypes specified the shapes and soft tissue anthropometrics of facial features. Each of the first generation of faces was rated by a subject (relative fitness measure) for beauty. The fittest genotypes then bred in proportion to their fitness, with crossover and mutation of the binary strings, to produce offspring which were again rated by the subject. This process continued until the most beautiful face, for that subject, was evolved. Forty Caucasian subjects (20 M, 20 F) were required to evolve their idealized beautiful female face using this procedure. The features and soft tissue anthropometrics of their final composites were compared to population norms. Also, the final composites, and different faces generated from the same data base, were rated for beauty by independent judges. The results support the conclusion that the concept of facial beauty is the result of sexual selection, and a beautiful female face has features and proportions indicative on high fertility.     

Osmoregulation or Turgor Regulation in Chara?

Chara corallina Klein ex Willd. wm. R.D.W. (= C. australis R. Br.), a fresh water alga, maintains a constant internal osmotic pressure when external osmotic pressure is increased. This results in a decrease in turgor pressure. Chara osmoregulates effectively in the presence of high CaCl₂ and raffinose, but is less efficient in response to increased NaCl. Decreasing external pH from 7 to 5 results in a decrease in turgor, but increasing it to values as high as 9 has no effect. Increasing the daily amount of light from 0.5 to 24 hours has no effect on turgor.     

Is there a gloxylate cycle in the liver of the fetal guinea pig?

Studies on the effect of the inhibitor of fatty acid oxidation (+)-octanoylcarnitine on the perfused liver of the 48–51 days fetal guinea pig indicate that the oxidation of endogenous fatty acids is a major source of carbon for the citric acid cycle and for synthesis of hexose. Consistent with this the liver can convert isocitrate to glyoxylate and glyoxylate to malate and may therefore operate a glyoxylate cycle allowing the net production of sugars from acetyl-CoA.     

Is there a pathology of prevention? the implications of visualizing the invisible in screening programs

The preventive orientation that has been gaining ground in Sweden is indicative of ways in which our society is organized to sustain values like a healthy life, a healthy body, and a healthy society. Search for health dangers and risks shows how medical technology has been integrated with our thinking about health. Preventive language, like all language of medicine, besides describing a pre-existing biological reality, creates in the process its own objects of analysis. This also has an impact and influences how lay people experience their bodies. The study presented focuses on one form of prevention in an attempt to describe how the ambition to secure a healthy society, through the detection of early disease, may have the opposite effect. Medical health-care ambitions in screening for cholesterolaemia will be related to implications for a group of men in whom cholesterol was found to be elevated. The men feel healthy yet are in some sense diseased. This raises the issue of visualizing the invisible in health care and the implications of such a process for the patients concermed.     

Is the involvement of opinion leaders in the implementation of research findings a feasible strategy?

Background

Although numerous therapies have been shown to be beneficial in the prevention of myocardial infarction and/or death in patients with coronary disease, these therapies are under-used and this gap contributes to sub-optimal patient outcomes. To increase the uptake of proven efficacious therapies in patients with coronary disease, we designed a multifaceted quality improvement intervention employing patient-specific reminders delivered at the point-of-care, with one-page treatment guidelines endorsed by local opinion leaders ("Local Opinion Leader Statement"). This trial is designed to evaluate the impact of these Local Opinion Leader Statements on the practices of primary care physicians caring for patients with coronary disease. In order to isolate the effects of the messenger (the local opinion leader) from the message, we will also test an identical quality improvement intervention that is not signed by a local opinion leader ("Unsigned Evidence Statement") in this trial.

Methods

Randomized trial testing three different interventions in patients with coronary disease: (1) usual care versus (2) Local Opinion Leader Statement versus (3) Unsigned Evidence Statement. Patients diagnosed with coronary artery disease after cardiac catheterization (but without acute coronary syndromes) will be randomly allocated to one of the three interventions by cluster randomization (at the level of their primary care physician), if they are not on optimal statin therapy at baseline. The primary outcome is the proportion of patients demonstrating improvement in their statin management in the first six months post-catheterization. Secondary outcomes include examinations of the use of ACE inhibitors, anti-platelet agents, beta-blockers, non-statin lipid lowering drugs, and provision of smoking cessation advice in the first six months post-catheterization in the three treatment arms. Although randomization will be clustered at the level of the primary care physician, the design effect is anticipated to be negligible and the unit of analysis will be the patient.

Discussion

If either the Local Opinion Leader Statement or the Unsigned Evidence Statement improves secondary prevention in patients with coronary disease, they can be easily modified and applied in other communities and for other target conditions.     

Is the preprophase band of microtubules a marker of organization in suspension cultures?

Summary To date it has been accepted that preprophase bands of microtubules (PPBs) either do not precede cell division or do so inconsistently in suspension cultures, the assumption being that such cultures proliferate in an unorganized state in which placement of cell plates is not regulated by the PPB system that is widespread in organized tissues. Using indirect immunofluorescence microscopy with antitubulin, the relative frequency of occurrence of PPBs in enzymatically separated cells from root tips and suspension cultures of carrot and tobacco, was quantified by taking the ratio of the number of PPBs: phragmoplast. This ratio was termed the PPB index.One carrot suspension culture proliferated in a medium containing 2,4-Dichlorophenoxyacetic acid (2,4-D), and recognizable stages in somatic embryogenesis formed when 2,4-D was removed from the medium. Another carrot suspension culture was nonembryogenic and removal of 2,4-D resulted in a reduction of cell division and increase in cell elongation. The tobacco culture was a cytokinin habituated cell line and also required 2,4-D to maintain cell division. It ceased proliferation, and cell elongation took place if 2,4-D was removed.The PPB index in the root tips from both species, and in both types of carrot suspension culture was approximately the same but was approx. 15-fold lower in the tobacco suspension. PPBs in the tobacco suspension were atypical in structure as well as sparse in numbers. The PPB index allows quantitative comparisons between different tissues to be made. The low PPB index and the irregular PPBs in the tobacco suspension correlates with its inability to undergo organized morphogenesis and generate spatially defined cell lineages upon 2,4-D removal. In contrast, the high PPB index in the carrot suspension cultures correlates with their potential for organized embryo formation, whether or not that potential is realized by withdrawal of 2,4-D. However, their high PPB index is not obligatorily coupled to embryogenesis.     

Thyroid Hormones Affect the Membrane Dipolar Organization. Is It a General Event in Their Non-genomic Action?

The surface balance technique was employed to study the interactions of 3,5,3',5' tetraiodo L-thyronine, 3,5,3' triiodo L-thyronine, and 3,5-diiodothyronine with monomolecular phospholipid monolayers spread at the air-water interface. With this technique the insertion of thyroid hormones into egg yolk phosphatidylcholine was investigated. An increase of surface pressure and a substantial decrement in surface potential were observed after the injection of these hormones beneath a phospholipid monolayer. The negative dipole contribution upon hormone interaction opposes the well-known positive contribution of phospholipids. These effects correlated with iodo content of the thyroid molecule analogues 3,5,3',5' tetraiodo L-thyronine >3,5,3' triiodo L-thyronine >3,5-diiodothyronine. To our knowledge, these observations suggest a new and surprising effect of thyroid hormones on the regulation of transmembrane dipolar organization.     

Is the meniscus of the knee joint a fibrocartilage?

A histological analysis of the structure of intact knee joint menisci was carried out in adult dogs. By means of specific histochemical methods for the connective tissue and cartilage, it was found that the meniscus as a whole does not have a unique structure. The anterior and posterior horns are populated by round chondroid cells encircled by abundant interstitial substance and branched wavy connective fibers; blood vessels are present. The outer third of the meniscus is constituted of cross bundles of connective fibers, fibrocytes and spindle-like areas of loose connective tissue with blood vessels. The inner avascular two thirds of the meniscus are filled with parallel circumferentially oriented fascicles of connective fibers, ovally elongated chondroid cells, and a small quantity of chondroid interstitial substance. In some menisci, in the inner two thirds of the body, there are isles of typical cartilage, which show metachromasia of the beta type and rarely of the gamma type. The occurrence and way of the manifestation of cartilage are of an individual character. The structural duality of the knee meniscus is accounted for by its functional duality manifested in offering resistance to the forces of traction and pressure, the latter ones favoring the process of evolution of tissue from connective, through chondroid, to cartilaginous.