Muscle cellular properties in the ice hockey player: a model for investigating overtraining?

In this study, we hypothesized that athletes involved in 5-6 months of sprint-type training would display higher levels of proteins and processes involved in muscle energy supply and utilization. Tissue was sampled from the vastus lateralis of 13 elite ice hockey players (peak oxygen consumption = 51.8 ± 1.3 mL·kg(-1)·min(-1); mean ± standard error) at the end of a season (POST) and compared with samples from 8 controls (peak oxygen consumption = 45.5 ± 1.4 mL·kg(-1)·min(-1)) (CON). Compared with CON, higher activities were observed in POST (p < 0.05) only for succinic dehydrogenase (3.32 ± 0.16 mol·(mg protein)(-1)·min(-1) vs. 4.10 ± 0.11 mol·(mg protein)(-1)·min(-1)) and hexokinase (0.73 ± 0.05 mol·(mg protein)(-1)·min(-1) vs. 0.90 ± 0.05mol·(mg protein)(-1)·min(-1)) but not for phosphorylase, phosphofructokinase, and creatine phosphokinase. No differences were found in Na(+),K(+)-ATPase concentration (β(max): 262 ± 36 pmol·(g wet weight)(-1) vs. 275 ± 27 pmol·(g wet weight)(-1)) and the maximal activity of the sarcoplasmic reticulum Ca(2+)-ATPase (98.1 ± 6.1 μmol·(g protein)(-1)·min(-1) vs. 102 ± 3.3 μmol·(g protein)(-1)·min(-1)). Cross-sectional area was lower (p < 0.05) in POST but only for the type IIA fibres (6312 ± 684 μm(2) vs. 5512 ± 335 μm(2)), while the number of capillary counts per fibre and the capillary to fibre area ratio were generally higher (p < 0.05). These findings suggest that elite trained ice hockey players display elevations only in support of glucose-based aerobic metabolism that occur in the absence of alterations in excitation-contraction processes.     

Stable karyotypes: a general rule for the fish of the family Prochilodontidae?

Cytogenetic studies involving the family Prochilodontidae have shown that these fish can be characterized by a constant diploid number and a conserved karyotypic macrostructure. This study focused on comparative physical chromosomal mapping using 18S and 5S rDNA to compare the species Semaprochilodus insignis and S. taeniurus. Our results indicated the conservation of large number of conventional chromosomal markers. The molecular cytogenetic analyses of the location of the 18S rDNA indicated the maintenance of a chromosome pair bearing these sites in both species analyzed, and it appears to be a conserved character among the majority of the species of this family. The stability of the number of 5S ribosomal DNA sites and their chromosomal localization as has been reported for the Prochilodontidae was not, however, confirmed for S. insignis and S. taeniurus, as these species showed multiple specific rDNA 5S sites. As such, and in spite of the fact that a number of studies indicate that the family Prochilodontidae has a conserved karyotypic structure, the utilization of molecular tools that use chromosomal segments as markers revealed that this presumed stability cannot be extended to the genome level for the species S. insignis and S. taeniurus.     

Fast methods of enantiopurity determination for the Soai reaction: Towards a general enantioenrichment detector?

The Soai reaction system possesses a remarkable combination of characteristics (enantioselective autocatalysis, strong positive nonlinearity leading to asymmetric amplification, ability to be triggered by wide variety of exogenous enantioenriched materials) that make it a potential starting point for a generalized detector for enantioenrichment. In this study we apply standard approaches used in pharmaceutical process research to the problem of developing a rapid method for analysis of the enantiopurity of the Soai reaction product. Several methods for rapid analysis (<1 min per sample) are described, including an approach using chiral supercritical fluid chromatography (SFC) and an approach using achiral chromatography with circular dichroism (CD) detection. Some thoughts on incorporation into a device for generalized enantioenrichment detection are presented.     

Probabilistic optimization of phenotype distributions: a general solution for the effects of uncertainty on natural selection?

Summary Explorations of optimizing selection often find discrepancies between the theoretically expected and observed phenotypes. Such discrepancies are usually attributed to a variety of potential constraints. We suggest that one common constraint, environmental uncertainty, may reduce the applicability of traditional deterministic or stochastic optimization methods and that many apparent discrepancies might be artifacts of these methods. Since natural selection is essentially a statistical process, we propose that a probabilistic optimization procedure, that includes all of the variability in phenotype distributions and associated fitness potential functions might offer better results. The traditional methods define an optimal gene or genotype as that which produces a phenotype distribution with a mean or other measure of central tendency that equals the value yielding the maximum fitness potential. Our method defines the optimal gene or genotype as that which produces the phenotype distribution that maximizes fitness summed or integrated over its associated fitness potential function. Often the central tendency of the phenotype distribution yielding the probabilistic optimum will differ from the deterministic expectation. This method is an extension of utility theory to any phenotypic character. We illustrate our method using an example based on Price and Waser's (1979) notion of optimal inbreeding via optimal pollen dispersal.     

Phylogeography of a nematode (Heligmosomoides polygyrus) in the western Palearctic region: persistence of northern cryptic populations during ice ages?

This study establishes the continental phylogeographical pattern of a European nematode, Heligmosomoides polygyrus (Dujardin, 1845; Heligmosomoidea). We sequenced 687 base pairs of the mitochondrial DNA (mtDNA) cyt b gene for 136 individuals collected in 22 localities. The results revealed that H. polygyrus populations are separated into five major units corresponding to the Italian, northern European (Denmark and Ireland), Iberian, western European, and Balkan populations. Different subclades were also observed within the first two groups. Based on the rate of molecular evolution of H. polygyrus cyt b gene-estimated to 3.5%-3.7% divergence per million years (Myr) in a previous study--the isolation time of the five clades was estimated between 2.5 +/- 0.24 and 1.5 +/- 0.23 million years ago. Moreover, H. polygyrus presents a higher genetic variability in the Mediterranean peninsulas as compared to northwestern Europe, highlighting the role of these regions as refuge areas. Like its specific host, the wood mouse Apodemus sylvaticus, H. polygyrus' pattern of postglacial recolonization of northwestern Europe was initiated from Iberian populations, while Italian and Balkan populations did not expand to the north. The results also suggest the existence of forested and temperate refuges in the southern British Isles during the Quaternary. Finally, the genetic diversity as well as the level of genetic divergence between the lineages of H. polygyrus are compared to those observed in other vertebrate and invertebrate phylogeographical studies: the existence of highly differentiated lineages in H. polygyrus (5%-10% of genetic divergence) highlights that the effects of Pleistocene climate changes on free-living organisms are also reflected in their obligate parasites.     

Identification of the mouse Loop‐tail gene: a model for human craniorachischisis?

Neural tube defects are one of the commonest human birth defects, with more than 0.5% of some populations affected. Mouse models are being used in an attempt to identify genes that could be involved in these malformations. Only two mouse mutations are known to lead to craniorachischisis, failure of closure of almost the entire neural tube. Two recent papers report that the gene for one of these, Loop-tail, has now been identified and sequenced.1, 2 It has been given the designation Ltap/Lpp1 and appears to function in floor plate formation. It will be of great interest to investigate the role of this gene in human neural tube defects.     

Oncogenic virus-associated neoplasia: a role for cyclin D1 genotypes influencing the age of onset of disease?

Cyclin D1 (CCND1) is a key regulatory protein at the G1/S checkpoint of the cell cycle. The purpose of our study was to assess the role of CCND1 genotypes influencing the age of onset of oncogenic virus-associated neoplasia. We conducted a hospital-based case-control study of 581 individuals, including 247 controls and 334 cases (108 nasopharyngeal and 226 cervical cancer cases). The polymorphism analysis was performed in blood samples by PCR-RFLP methodology. Age-adjusted logistic regression analysis indicates that individuals carrying two G-alleles have an increased genetic susceptibility for the development of oncogenic virus-associated cancers (aOR = 2.02, 95% CI 1.30-3.14, P = 0.002). Moreover, our results indicate that the waiting time for onset of oncogenic virus-associated neoplasia in patients homozygous (GG) for CCND1 genotypes (52 years) was 12 years earlier in comparison with patients carrying AG or AA genotypes (60 years) (log-rank test: P = 0.0003). Our results may be important in contributing to a more extensive knowledge of the mechanisms involved in oncogenic virus-associated carcinogenesis, as CCND1 may be an important target for the development of new strategies for cancer treatment and prevention.     

Aggregation and collapse of fungal wall vesicles in hyphal tips: a model for the origin of the Spitzenk?rper

The intracellular origins of polarity and branch initiation in fungi centre upon a localization in the supply of fungal wall constituents to specific regions on the hyphal wall. Polarity is achieved and maintained by accumulating secretory vesicles, prior to incorporation into the wall, in the form of an apical body or Spitzenkörper. However, neither the mechanisms leading to this accumulation nor the initiation of branching, are as yet understood. We propose a mechanism, based on experimental evidence, which considers the mechanical properties of the cytoskeleton in order to explain these phenomena. Cytoskeletal viscoelastic forces are hypothesized to be responsible for biasing vesicles in their motion, and a mathematical model is derived to take these considerations into account. We find that, as a natural consequence of the assumed interactions between vesicles and cytoskeleton, wall vesicles aggregate in a localized region close to the tip apex. These results are used to interpret the origin of the Spitzenkörper. The model also shows that an aggregation peak can collapse and give rise to two new centres of aggregation coexisting near the tip. We interpret this as a mechanism for apical branching, in agreement with published observations. We also investigate the consequences and presumptive role of vesicle–cytoskeleton interactions in the migration of satellite Spitzenkörper. The results of this work strongly suggest that the formation of the Spitzenkörper and the series of dynamical events leading to hyphal branching arise as a consequence of the bias in vesicle motion resulting from interactions with the cytoskeleton.     

Transport physiology of the urinary bladder in teleosts: a suitable model for renal urea handling?

The transport physiology of the urinary bladder of both the freshwater rainbow trout (Oncorhychus mykiss) and the marine gulf toadfish (Opsanus beta) was characterized with respect to urea, and the suitability of the urinary bladder as a model for renal urea handling was investigated. Through the use of the in vitro urinary bladder sac preparation urea handling was characterized under control conditions and in the presence of pharmacological agents traditionally used to characterize urea transport such as urea analogues (thiourea, acetamide), urea transport blockers (phloretin, amiloride), and hormonal stimulation (arginine vasotocin; AVT). Na(+)-dependence and temperature sensitivity were also investigated. Under control conditions, the in vitro trout bladder behaved as in vivo, demonstrating significant net reabsorption of Na(+), Cl(-), water, glucose, and urea. Bladder urea reabsorption was not affected by pharmacological agents and, in contrast to renal urea reabsorption, was not correlated to Na(+). However, the trout bladder showed a threefold greater urea permeability compared to artificial lipid bilayers, a prolonged phase transition with a lowered E(a) between 5 degrees C and 14 degrees C, and differential handling of urea and analogues, all suggesting the presence of a urea transport mechanism. The in vitro toadfish bladder did not behave as in vivo, showing significant net reabsorption of Na(+) but not of Cl(-), urea, or water. As in the trout bladder, pharmacological agents were ineffective. The toadfish bladder showed no differential transport of urea and analogues, consistent with a low permeability storage organ and intermittent urination. Our results, therefore, suggest the possibility of a urea transport mechanism in the urinary bladder of the rainbow trout but not the gulf toadfish. While the bladders may not be suitable models for renal urea handling, the habit of intermittent urination by ureotelic tetrapods and toadfish seems to have selected for a low permeability storage function in the urinary bladder.     

Tracheobronchial air-liquid interface cell culture: a model for innate mucosal defense of the upper airways?

Human tracheobronchial epithelial cells grown in air-liquid interface culture have emerged as a powerful tool for the study of airway biology. In this study, we have investigated whether this culture system produces "mucus" with a protein composition similar to that of in vivo, induced airway secretions. Previous compositional studies of mucous secretions have greatly underrepresented the contribution of mucins, which are major structural components of normal mucus. To overcome this limitation, we have used a mass spectrometry-based approach centered on prior separation of the mucins from the majority of the other proteins. Using this approach, we have compared the protein composition of apical secretions (AS) from well-differentiated primary human tracheobronchial cells grown at air-liquid interface and human tracheobronchial normal induced sputum (IS). A total of 186 proteins were identified, 134 from AS and 136 from IS; 84 proteins were common to both secretions, with host defense proteins being predominant. The epithelial mucins MUC1, MUC4, and MUC16 and the gel-forming mucins MUC5B and MUC5AC were identified in both secretions. Refractometry showed that the gel-forming mucins were the major contributors by mass to both secretions. When the composition of the IS was corrected for proteins that were most likely derived from saliva, serum, and migratory cells, there was considerable similarity between the two secretions, in particular, in the category of host defense proteins, which includes the mucins. This shows that the primary cell culture system is an important model for study of aspects of innate defense of the upper airways related specifically to mucus consisting solely of airway cell products.     

Can hybridization cause local extinction: a case for demographic swamping of the Australian native Senecio pinnatifolius by the invasive Senecio madagascariensis?

Hybridization between native and invasive species can have several outcomes, including enhanced weediness in hybrid progeny, evolution of new hybrid lineages and decline of hybridizing species. Whether there is a decline of hybridizing species largely depends on the relative frequencies of parental taxa and the viability of hybrid progeny. Here, the individual- and population-level consequences of hybridization between the Australian native Senecio pinnatifolius and the exotic Senecio madagascariensis were investigated with amplified fragment length polymorphism (AFLP) markers, and this information was used to estimate the annual loss of viable seeds to hybridization. A high frequency (range 8.3-75.6%) of hybrids was detected in open pollinated seeds of both species, but mature hybrids were absent from sympatric populations. A hybridization advantage was observed for S. madagascariensis, where significantly more progeny than expected were sired based on proportional representation of the two species in sympatric populations. Calculations indicated that S. pinnatifolius would produce less viable seed than S. madagascariensis, if hybridization was frequency dependent and S. madagascariensis reached a frequency of between 10 and 60%. For this native-exotic species pair, prezygotic isolating barriers are weak, but low hybrid viability maintains a strong postzygotic barrier to introgression. As a result of asymmetric hybridization, S. pinnatifolius would appear to be under threat if S. madagascariensis increases numerically in areas of contact.     

Optimal combination of theory and experiment for the characterization of the protein folding landscape of S6: how far can a minimalist model go?

The detailed characterization of the overall free energy landscape associated with the folding process of a protein is the ultimate goal in protein folding studies. Modern experimental techniques provide accurate thermodynamic and kinetic measurements on restricted regions of a protein landscape. Although simplified protein models can access larger regions of the landscape, they are oftentimes built on assumptions and approximations that affect the accuracy of the results. We present a new methodology that allows to combine the complementary strengths of theory and experiment for a more complete characterization of a protein folding landscape. We prove that this new procedure allows a simplified protein model to reproduce remarkably well (correlation coefficient > 0.9) all experimental data available on free energies differences upon single mutations for S6 ribosomal protein and two circular permutants. Our results confirm and quantify the hypothesis, recently formulated on the basis of experimental data, that the folding landscape of protein S6 is strongly affected by an atypical distribution of contact energies.     

Immunotherapy of cancer for the elderly patient: does allogeneic bone marrow transplantation after nonmyeloablative conditioning provide a new option?

The critical role of antigen-specific T cells in cancer immunotherapy has been amply demonstrated. Though success of clinical trials still remains far behind expectations, the continuous improvement in our understanding of the biology of the immune response will provide the basis for optimized cancer vaccines. This review focuses on active therapeutic vaccination after allogeneic bone marrow cell transplantation with nonmyeloablative conditioning. This approach could provide a major breakthrough in cancer immunotherapy, particularly of elderly patients. The senescent immune system, mainly the T-cell compartment, displays reduced responsiveness, and this has to be overcome if therapeutic vaccination is to be of benefit for the patient. Although the defects are quite well characterized, the inducing factors and ways to overcome them are still to be explored in more detail. Many questions also remain to be answered in the field of allogeneic bone marrow transplantation after nonmyeloablative conditioning to optimize this therapeutic setting in cancer immunotherapy. Current considerations to improve engraftment and to reduce graft-versus-host disease while strengthening graft-versus-tumor reactivity will be briefly reviewed. Finally, I will discuss whether tumor-reactive T cells can be naturally maintained during the process of T-cell maturation in the allogeneic host. Provided this hypothesis can be substantiated, a T-cell vaccine will meet a pool of virgin T cells in the allogeneically reconstituted host, which are tolerant toward the host but not anergized toward tumor antigens presented by MHC molecules of the host. Inevitably, the problem of the aged immune system would be circumvented.Abbreviations APC antigen-presenting cell - BMCT bone marrow cell transplantation - CTL cytotoxic T cell - DC dendritic cell - GvHD graft-versus-host disease - GvT graft versus tumor - HvG host-versus-graft - LAK lymphokine-activated killer cell - mAB monoclonal antibody - MHC major histocompatibility complex - TCR T-cell receptor - TH helper T cell - TIL tumor-infiltrating leukocyteM. Zöller was supported by the Tumorzentrum Heidelberg/Mannheim, the Mildred-Scheel-Stiftung für Krebsforschung, the José Carreras Leukemia Foundation, and a German-Israel Joint Program.     

The genetic legacy of lower sea levels: does the confluence of rivers during the last glacial maximum explain the contemporary distribution of a primary freshwater fish (Pseudobarbus burchelli, Cyprinidae) across isolated river systems?

The primary freshwater fish Pseudobarbus burchelli (Smith 1841) occurs across four presently isolated river systems in the south-western cape floristic region of South Africa. Mitochondrial DNA cytochrome b (701 base pairs) and control region (601 base pairs) genes were sequenced to assess the evolutionary history of P. burchelli and evaluate the role of climatic and landscape changes in shaping patterns of genetic variation in this species. We identified three historically isolated lineages in P. burchelli: a widespread lineage that occurs across three isolated river systems and two geographically restricted lineages. The results were evaluated against predictions of the confluence of river systems during low sea levels of the last glacial maximum. Occurrence of the widespread Breede lineage in the Duiwenhoks River system is consistent with reconstructed palaeoriver systems. However, the occurrence of this lineage in the Goukou river system that formed part of the eastern Gourits–Goukou palaeoriver system can only be explained by translocation or a recent river capture or episodic inundation of low drainage divides. Extreme ecological gradients or the potential presence of instream physical barriers could have prevented an exchange of lineages between the Breede and Heuningnes river systems.     

New methods to analyse auxin-induced growth II: The swelling reaction of protoplasts – a model system for the analysis of auxin signal transduction?

A method for monitoring the time course of auxin-induced volume changesby protoplasts at a high temporal resolution was developed for Zeamays coleoptile protoplasts. Auxins, like indole-3-acetic acid(IAA), induce a rapid change in volume. Immediately after addition ofthis auxin, a transient shrinkage was observed, followed by a long-termswelling response. This reaction occurred in the same time window as thetypical auxin growth response of intact coleoptiles. Active auxins, like1-naphthalene acetic acid (1-NAA) and 4-chloroindole-3-acetic acid(4-Cl-IAA), caused similar volume changes, whereas the inactive analogue2-naphthalene acetic acid (2-NAA) had no effect. The phytotoxinfusicoccin (FC) induced a rapid swelling response. We conclude that thissingle cell system is very adequate to analyse mechanisms of auxinsignal transduction.     

Monoamines activate neuropeptide signaling cascades to modulate nociception in C. elegans: a useful model for the modulation of chronic pain?

Monoamines and neuropeptides interact to modulate key behaviors in most organisms. This review is focused on the interaction between octopamine (OA) and an array of neuropeptides in the inhibition of a simple, sensory-mediated aversive behavior in the C. elegans model system and describes the role of monoamines in the activation of global peptidergic signaling cascades. OA has been often considered the invertebrate counterpart of norepinephrine, and the review also highlights the similarities between OA inhibition in C. elegans and the noradrenergic modulation of pain in higher organisms.     

Disruption of the blood-testis barrier integrity by bisphenol A in vitro: Is this a suitable model for studying blood-testis barrier dynamics?

Bisphenol A, an estrogenic environmental toxicant, has been implicated to have hazardous effects on reproductive health in humans and rodents. However, there are conflicting reports in the literature regarding its effects on male reproductive function. In this study, it was shown that in adult rats treated with acute doses of bisphenol A, a small but statistically insignificant percentage of seminiferous tubules in the testes displayed signs of germ cell loss, consistent with some earlier reports. It also failed to disrupt the blood-testis barrier in vivo. This is possibly due to the low bioavailability of free bisphenol A in the systemic circulation. However, bisphenol A disrupted the blood-testis barrier when administered to immature 20-day-old rats, consistent with earlier reports concerning the higher susceptibility of immature rats towards bisphenol A. This observation was confirmed using primary Sertoli cells cultured in vitro with established tight junction-permeability barrier that mimicked the blood-testis barrier in vivo. The reversible disruption of Sertoli cell tight junction barrier by bisphenol A was associated with an activation of ERK, and a decline in the levels of selected proteins at the tight junction, basal ectoplasmic specialization, and gap junction at the blood-testis barrier. Studies by dual-labeled immunofluorescence analysis and biotinylation techniques also illustrated declining levels of occludin, connexin 43, and N-cadherin at the cell–cell interface following bisphenol A treatment. In summary, bisphenol A reversibly perturbs the integrity of the blood-testis barrier in Sertoli cells in vitro, which can also serve as a suitable model for studying the dynamics of the blood-testis barrier.