A monitoring,feedback, and triggering system for reproducible breath-hold MR imaging

A technique is described that provides improved reproducibility of breath-holding for MR image acquisition by monitoring the superior-inferior (S/I) position of the diaphragm. The method incorporates detection of the level of inspiration using an MR signal, rapid display to the patient of diaphragm position to enable breath-hold adjustment, and triggering of image data acquisition once appropriate position is attained. The response time of the system is short, approximately 10 ms. Studies in six volunteers using this method demonstrate a considerable decrease in the S/I range of diaphragm position over 10 consecutive periods of suspended respiration. The mean range is 1.3 mm with the system, while it is 8.3 mm without using it is expected that this method will be of assistance in many abdominal and cardiothoracic studies that use breath-hold techniques.     

Preparation of metaphase chromosomes in cell cultures derived from an aquatic reptile

Summary Cell cultures (GTS) of epithelioid nature derived from the skin of a green sea turtle,Chelonia mydas, were treated with colchicine at a final concentration of 0.5 g/ml for 16 h. Mitotic cells were harvested by brief treatment with trypsin-Versene, subjected to hypotonic solution (1% sodium citrate) and fixed in (13) acetic acid: methanol. Giemsa stained preparations were photographed on High Contrast Copy film under phase contrast optics using a bluegreen filter. The result was significant enhancement of the microchromosome portion of the complement morphologically characteristic of reptilian metaphase chromosomes. By this method it was determined that the GTS cell line retains the female diploid number of the Chelonia species.     

Relating cell and tissue mechanics: Implications and applications

The Differential Adhesion Hypothesis (DAH) posits that differences in adhesion provide the driving force for morphogenetic processes. A manifestation of differential adhesion is tissue liquidity and a measure for it is tissue surface tension. In terms of this property, DAH correctly predicts global developmental tissue patterns. However, it provides little information on how these patterns arise from the movement and shape changes of cells. We provide strong qualitative and quantitative support for tissue liquidity both in true developmental context and in vitro assays. We follow the movement and characteristic shape changes of individual cells in the course of specific tissue rearrangements leading to liquid-like configurations. Finally, we relate the measurable tissue-liquid properties to molecular entities, whose direct determination under realistic three-dimensional culture conditions is not possible. Our findings confirm the usefulness of tissue liquidity and provide the scientific underpinning for a novel tissue engineering technology. Developmental Dynamics 237:2438-2449, 2008. (c) 2008 Wiley-Liss, Inc.     

Structural changes induced in scallop heavy meromyosin molecules by Ca2+ and ATP

Summary We have used physicochemical and ultrastructural methods to investigate the effects of Ca²⁺ and ATP on the structure of purified heavy meromyosin (HMM) from the striated adductor muscle of the scallop, a species with myosin-linked regulation. Using papain as a structural probe, we found that, in the presence of ATP, the head/tail junction was five times more susceptible to digestion at high levels of Ca²⁺ than at low levels.wBy HPLC gel filtration, two fractions of scallop HMM with different Stokes radii were detected in the presence of ATP at low Ca²⁺, while at high Ca²⁺ a single peak with the larger Stokes radius predominated. Electron microscopy of rotary-shadowed HMM suggested that molecules with the smaller Stokes radius had their heads bent back towards their tails, while those with the larger radius had heads pointing away from the tail. The number of molecules with their heads bent back decreased at high Ca²⁺ levels. The data also showed that in the absence of ATP or at high salt, HMM molecules behaved similarly to those in the presence of ATP at high Ca²⁺.These results suggest that scallop myosin heads can exist in two conformations (heads down towards the tail and heads up away from the tail) and that the equilibrium between these two conformations is altered by the concentrations of salt, ATP and Ca²⁺. However, the equilibrium between the two forms appears to be too slow to be involved in regulating contraction. The heads-down configuration may instead be related to the inactive, folded (10S) form of scallop myosin and possibly involved in filament assembly during development.     

Effect of amiloride on electrolyte concentrations and rubidium uptake in principal and mitochondria-rich cells of frog skin

The role of mitochondria-rich cells (MR cells) in transepithelial Na transport was investigated by determining electrolyte concentrations and Rb uptake in individual cells of frog skin epithelium using electron microprobe analysis. Measurements were performed under control conditions and after blocking the transepithelial Na transport with amiloride. Under control conditions, Na and Cl concentrations of MR cells scattered much more than those of principal cells and ranged from a few up to more than 30 mmol/kg wet weight. Rb uptake from the basal side into individual MR cells also showed a large variation and was, on the average, much less pronounced than into the principal cells. In principal cells, amiloride reduced the Na concentration and Rb accumulation. In contrast, no effect was observed upon electrolyte concentration and Rb uptake of MR cells. Rb uptake was correlated to the Na concentration of MR cells both under control conditions and after amiloride. It is concluded that, in contrast to the principal cells, MR cells are not involved in amiloride-sensitive transepithelial Na transport and that their Na/K-pump activity is very low.     

Mechanisms for differential block among single myelinated and non-myelinated axons by procaine

1. The differential sensitivity of saphenous nerve fibres in the cat to block by procaine HCl was re-examined by recording identifiable unit action potentials from small nerve filaments.2. Small myelinated axons were blocked more quickly than large myelinated axons, but this differential effect could not be accounted for by differences in anaesthetic concentration requirements.3. The onset of block in non-myelinated axons was slower than or equal to that of small myelinated axons depending on anaesthetic concentration.4. Absolute differential block of non-myelinated and small myelinated axons was obtained by limiting the length of axons exposed to procaine to 2 mm.5. Differential rates of blocking among myelinated axons appear to depend on differences in the length of axons that must be exposed to blocking concentrations of procaine and to arise from the irregular distribution of such concentrations within an exposed nerve.     

A unique exonic splice enhancer mutation in a family with X-linked mental retardation and epilepsy points to a novel role of the renin receptor

The renin-angiotensin system (RAS) is essential for blood pressure control and water-electrolyte balance. Until the discovery of the renin receptor, renin was believed to be mainly a circulating enzyme with a unique function, the cleavage of angiotensinogen. We report a unique mutation in the renin receptor gene (ATP6AP2) present in patients with X-linked mental retardation and epilepsy (OMIM no. 300423), but absent in 1200 control X-chromosomes. A silent mutation (c.321C>T, p.D107D) residing in a putative exonic splicing enhancer site resulted in inefficient inclusion of exon 4 in 50% of renin receptor mRNA, as demonstrated by quantitative RT-PCR. Analysis of membrane associated-receptor molecular forms showed the presence of full-length and truncated proteins in the patient. Functional analysis demonstrated that the mutated receptor could bind renin and increase renin catalytic activity, similar to the wild-type receptor, but resulted in a modest and reproducible impairment of ERK1/2 activation. Thus, our findings confirm the importance of the RAS in cognitive processes and indicate a novel specific role for the renin receptor in cognitive functions and brain development.     

Randomised controlled trial of pelvic floor muscle exercises and manometric biofeedback for erectile dysfunction.

BACKGROUND: The pelvic floor muscles are active in normal erectile function. Therefore, it was hypothesised that weak pelvic floor muscles could be a cause of erectile dysfunction.AIMS: To compare the efficacy of pelvic floor muscle exercises and manometric biofeedback with lifestyle changes for men with erectile dysfunction.Design of study: Randomised controlled trial.SETTING: The Somerset Nuffield Hospital, Taunton, United Kingdom.METHOD: Fifty-five men with erectile dysfunction (median age 59.2 years; range 22-78 years) were enrolled from a local urology clinic. Of these, 28 participants were randomised to an intervention group and engaged in pelvic floor exercises, as well as receiving biofeedback and suggestions for lifestyle changes. Twenty-seven controls were solely advised on lifestyle changes. Baseline, 3- and 6-month assessments were: erectile function domain of International Index of Erectile Function (IIEF), Partner's International Index of Erectile Function (PIIEF), Erectile Dysfunction-Effect on Quality of Life (ED-EQoL), anal manometry, digital anal measurements, and clinical assessment by an assessor blind to treatment allocation. After 3 months, the control group were transferred to the active arm.RESULTS: At 3 months, compared with controls, men in the intervention group showed significant mean increases in the erectile function domain of the IIEF (6.74 points, P = 0.004); anal pressure (44.16 cmH(2)O, P <0.001); and digital anal grades (1.5 grades, P <0.001). All showed further improvement in these outcomes at 6 months. Similar benefits were seen in men of the control arm after transfer to active treatment. A total of 22 (40.0%) participants attained normal function, 19 (34.5%) participants had improved erectile function, and 14 (25.5%) participants failed to improve.CONCLUSION: Pelvic floor muscle exercises and biofeedback are an effective treatment for men with erectile dysfunction.