Structural insight into M-band assembly and mechanics from the titin-obscurin-like-1 complex

In the sarcomeric M-band, the giant ruler proteins titin and obscurin, its small homologue obscurin-like-1 (obsl1), and the myosin cross-linking protein myomesin form a ternary complex that is crucial for the function of the M-band as a mechanical link. Mutations in the last titin immunoglobulin (Ig) domain M10, which interacts with the N-terminal Ig-domains of obscurin and obsl1, lead to hereditary muscle diseases. The M10 domain is unusual not only in that it is a frequent target of disease-linked mutations, but also in that it is the only currently known muscle Ig-domain that interacts with two ligands—obscurin and obsl1—in different sarcomeric subregions. Using x-ray crystallography, we show the structural basis for titin M10 interaction with obsl1 in a novel antiparallel Ig-Ig architecture and unravel the molecular basis of titin-M10 linked myopathies. The severity of these pathologies correlates with the disruption of the titin-obsl1/obscurin complex. Conserved signature residues at the interface account for differences in affinity that direct the cellular sorting in cardiomyocytes. By engineering the interface signature residues of obsl1 to obscurin, and vice versa, their affinity for titin can be modulated similar to the native proteins. In single-molecule force-spectroscopy experiments, both complexes yield at forces of around 30 pN, much lower than those observed for the mechanically stable Z-disk complex of titin and telethonin, suggesting why even moderate weakening of the obsl1/obscurin-titin links has severe consequences for normal muscle functions.     

An automatic measure of progression during colonoscopy correlates to patient experienced pain

Background: Colonoscopy screening and surveillance programs depend on patient’s tolerable experience, which is associated with competence of the endoscopist. The Colonoscopy Progression Score (CoPS) is an automated tool based on recording of the Magnetic Scope Imager (MEI) picture in order to track progression. CoPS deliver a numeric score and a graphic map. A high score expresses a rapid and smooth progression. Aims of study were to explore the correlation between CoPS and patient experienced pain and to identity locations associated with pain.

Methods and materials: Patients listed for colonoscopy were included and asked to reply to pain by pressing a rubber ball. The signal was recorded simultaneous to CoPS. Patients evaluated the experience on a Visual Analogue Scale (VAS). CoPS and recorded pain events were used to create a pain sensitive CoPS-map (S-CoPS map).

Results: A total of 58 complete recordings were used for evaluation. We demonstrated a moderate correlation between CoPS and patient experienced pain, Pearson’s r?=??0.47 (p?Conclusion: A moderate correlation between CoPS and patient experienced pain suggest that CoPS measure inserting skills but might also be a measure of a gentle performance. The graphic S-CoPS-map can be used to point-out painful passages and aid planning of future colonoscopies.     

Biosimulation Modeling for Diabetes: Contributions of Mathematical Modeling of Beta Cells to the Understanding of Beta-Cell Oscillations and Insulin Secretion

Mathematical modeling of pancreatic beta cells has contributed significantly to the understanding of the mechanisms involved in glucose-stimulated insulin secretion (GSIS). Early models of insulin secretion built in the 1970s were phenomenological with little biological foundation for the proposed mechanisms. In the 1980s, models focused on identifying the regulation of bursting electrical activity known to be important for insulin secretion. The main result was to reject proposed mechanisms as new data emerged, but important results of the role of cell-to-cell coupling were also established. New models have been proposed that provide possible explanations for the occurrence of various patterns of bursting and calcium oscillations. In addition, modeling has played an important role in comparing competing effects of calcium on both NADH and adenosine 3′-5′-cyclic monophosphate levels. Models including modern cell biological results of the regulation of insulin containing granules and cell heterogeneity have appeared, providing updated versions of the early models proposed in the 1970s. These models, when coupled to electrophysiological- and calcium-based ones, have the prospect to aid in understanding the overall picture of GSIS. In addition, they might be useful for estimating in vivo beta-cell functioning. Beta-cell modeling will likely move closer to clinical applications, where it can be expected to play an important role, as it has and will, in understanding the complex oscillatory phenomena observed in beta cells and islets.     

Nitric oxide synthase type-1 modulates cardiomyocyte contractility and calcium handling: association with low intrinsic aerobic capacity.

BACKGROUND: The neuronal isoform of nitric oxide synthase (NOS-1) may be an important regulator of cardiac contractility by modifying calcium release and uptake from sarcoplasmic reticulum. Our working hypothesis was that NOS-1 modulates cardiomyocyte contractility more markedly in rat lines with low versus high congenital aerobic fitness. METHODS AND RESULTS: Rats performed high-intensity interval treadmill running 5 days per week over 8 weeks; age-matched sedentary rats served as controls. At baseline before the training program, aerobic fitness measured as maximal oxygen uptake was 30% higher, and cardiomyocyte contractility measured as fractional shortening 42% higher in high than in low congenital aerobic fitness rats. Training markedly increased aerobic fitness as well as cardiomyocyte contractility, relaxation and corresponding changes in calcium transient in both lines. Selective inhibition of NOS-1 increased cardiomyocyte contractility (12-43%) and calcium transient amplitude (10-28%), prolonged time to 50% relengthening (13-52%) and time to 50% calcium decay (17-35%), in all groups. Interestingly, NOS-1-inhibition abolished the difference in systolic events between low and high congenital aerobic fitness whereas no such findings occurred in diastolic parameters. CONCLUSION: NOS-1-derived nitric oxide production is a modulator of cardiomyocyte contractile performance and calcium handling in rats. It accounts for some of the difference between rats with low versus high congenital aerobic fitness, whereas it contributes little during adaptation to exercise training.     

The disease modifying osteoarthritis drug (DMOAD): Is it in the horizon?

Till date, the pharmaceutical industry has failed to bring effective and safe disease modifying osteoarthritic drugs (DMOADs) to the millions of patients suffering from this serious and deliberating disease. We provide a review of recent data reported on the investigation of DMOADs in clinical trials, including compounds inhibiting matrix-metalloproteinases (MMPs), bisphosphonates, cytokine blockers, calcitonin, inhibitors of inducible nitric oxide synthase (iNOS), doxycycline, glucosamine, and diacereine. We discuss the challenges associated with the drug development process in general and with DMOADs in particular, and we advance the need for a new development paradigm for DMOADs. Two central elements in this paradigm are a stronger focus on the biology of the joint and the application of new and more sensitive biomarkers allowing redesign of clinical trials in osteoarthritis.     

Adult Refsum Disease: A Form of Tapetoretinal Dystrophy Accessible to Therapy

Adult Refsum disease is characterized by an elevated plasma phytanic acid level and high concentrations of phytanic acid in a variety of tissues. Besides tapetoretinal degeneration, additional symptoms are anosmia, skeletal malformations, chronic polyneuropathy, cerebellar ataxia, sensorineural hearing loss, ichthyosis, and cardiac abnormalities. A diet low in phytanic acid ameliorates polyneuropathy and ataxia and slows or even stops the other manifestations. In order to be able to apply dietary therapy, as many patients as possible (even better if all of them are) have to be identified at an early stage. The ophthalmologist plays a crucial role in achieving this goal because of the early manifestation of the tapetoretinal degeneration.     

Co-operation and conflict in a hospital: interprofessional differences in perception and management of conflicts

This article presents a case study of a Norwegian hospital, analysing how health professionals manage conflicts related to work co-operation. Alltogether, 29 health professionals working in the hospital were interviewed, and data was analysed according to a grounded theory approach. When in conflict, health professionals seem to use three major approaches to handling the situation: avoidance, forcing and negotiation, and usually in that order. Avoidance behaviour or suppression is the most common reaction to an emerging conflict. If the use of power does not re-establish a balance between the participants, one negotiates. These conflict styles seem to be determined by two major factors: the perceived interdependence between parties and the perceived urgency of doing something about the situation. Nurses and physicians in particular seem to differ considerably in their perception of what is a conflict and when to do something about it. Such differences in perceptions and the extensive use of avoidance represent important challenges to managers and clinical leaders when it comes to advancing interprofessional co-operation.     

Cysteine uptake by Saccharomyces cerevisiae is accomplished by multiple permeases

Uptake by Saccharomyces cerevisiae of the sulphur-containing amino acid L-cysteine was found to be non-saturable under various conditions, and uptake kinetics suggested the existence of two or more transport systems in addition to the general amino-acid permease, Gap1p. Overexpression studies identified BAP2, BAP3, AGP1 and GNP1 as genes encoding transporters of cysteine. Uptake studies with disruption mutants confirmed this, and identified two additional genes for transporters of cysteine, TAT1 and TAT2, both very homologous to BAP2, BAP3, AGP1 and GNP1. While Gap1p and Agp1p appear to be the main cysteine transporters on the non-repressing nitrogen source proline, Bap2p, Bap3p, Tat1p, Tat2p, Agp1p and Gnp1p are all important for cysteine uptake on ammonium-based medium. Furthermore, whereas Bap2p, Bap3p, Tat1p and Tat2p seem most important under amino acid-rich conditions, Agp1p contributes significantly when only ammonium is present, and Gnp1p only contributes under the latter condition. Received: 25 January / 15 March 1999