Actions of the phorbol ester, PMA, on ATP-induced fluctuations in cytoplasmic calcium in single isolated bovine aortic endothelial cells.

Agonist-evoked rises in [Ca2+]i were recorded from single bovine aortic endothelial cells in the middle of confluent monolayers by dual-wavelength microspectrofluorimetry. Low doses of ATP (1-5 microM) induced a transient rise in [Ca2+]i followed by a maintained plateau phase upon which were superimposed irregular fluctuations in [Ca2+]i. The mechanism underlying these fluctuations is not known. Addition of the phorbol ester, phorbol 12-myristate 13-acetate (PMA), to single cells displaying ATP-induced fluctuations reduced the amplitude and frequency of these fluctuations but the maintained plateau phase was unaffected. Elevation of intracellular cAMP by activation of adenylate cyclase with forskolin, or application of dibutyryl cAMP did not affect the ATP-induced fluctuations. These results suggest a possible role for the diacylglycerol limb of the phosphoinositide hydrolysis pathway, via activation of protein kinase C, but not cAMP, in the mechanism responsible for generating ATP-induced fluctuations in [Ca2+]i.     

The effect of pH on the in vitro colony forming ability of transitional cell carcinoma cells treated with various chemotherapeutic agents: implications for in vivo therapy.

Extracellular pH may affect the sensitivity of cancer cells to chemotherapy agents. In an attempt to maximize the conditions for chemotherapy treatment of transitional cell carcinoma we tested the effect of pH on sensitivity of MGH-U3 transitional cell carcinoma cell line to thiotepa, doxorubicin, and mitomycin c in vitro. The toxicity of each agent tested varied with pH. There was no variation in cell growth in response to pH alone. The cytotoxic activity of thiotepa was markedly enhanced when cells were treated with a diluent pH of 5.5. Significant differences were also observed after treatment with doxorubicin and mitomycin c with a diluent pH of 7.0. This in vitro assay may be useful for clinical application of pH modulation during intravesical chemotherapy.     

FFT-based digital tactile vocoder system for real-time use

A microprocessor-based real-time digital vibrotactile vocoder system has been developed to train the deaf and for artificial hearing research. The system is composed of a microcomputer module with a digital signal processor interface units and an attenuator/driver circuit. Live or digitised (stored or synthetic) speech is presented to the skin spectrally through a belt housing eight or 16 vibrators. Speech is processed in real time using a fast Fourier transform. The system is also capable of presenting any arbitrary spatiotemporal pattern on the skin for artificial hearing experiments. A preliminary experiment with a deaf subject indicates that the system is potentially an effective device for artificial hearing.     

What is the most useful and cost-effective strategy to screen for left ventricular systolic dysfunction in clinical practice?: reply

We agree with many of the comments made by Yamada et al.,¹namely that the ECG and NTproBNP are fundamental tests usedin the diagnosis of heart failure and that     

Examining preferences for allocating health care gains

This study is part of a programme to elicit and examine community preferences for health care in different contexts. Data were obtained from a group of predominantly Australian health care decision-makers. A short questionnaire contained six valuation questions and four demographic questions. The six valuation questions posed choices where equal health gains were to be allocated to different population groups based upon: age; sex; current health; socio-economic status; across time; and across different numbers of individuals. The results provide some evidence that respondents were prepared to discriminate between health gains derived in different contexts especially where health gains were to be allocated between groups of different health status and over time. Further research is planned and the possible implications for health policy, and in particular for resource allocation in health care, are briefly discussed.     

ADAMTS-4 activity on a proteoglycan vs. a polypeptide is controlled by the ancillary domains

Introduction In combination with the catalytic domain, the ancillary domains of the ADAMTS' are proposed to regulate activity via interactions with sulfated GAGs, the extracellular matrix (ECM) and cell surface. Interactions with both GAGs and the ECM have been attributed to the thrombospondin (TSP) type 1 motifs and spacer region ( Kuno and Matsushima 1998 ; Flannery et al. 2002 ). ADAMTS‐1, ‐4 and ‐5, all undergo cleavage within their respective spacer regions ( Flannery et al. 2002 ; Rodriguez‐Manzaneque et al. 2000 ; Georgiadis et al. 2002 ), an event which has been reported to increase activity towards the interglobular domain of aggrecan and decrease the heparin affinity of ADAMTS‐4 ( Flannery et al. 2002 ; Gao et al. 2002 ). Materials and methods V5‐ and His‐tagged recombinant human ADAMTS‐4 constructs terminating after the catalytic (?DTS), disintegrin‐like (?TS), TSP (?S) or spacer region (Full) were expressed in High‐Five cells. Proteoglycanase activities of the resultant proteins were assayed with solution digests of aggrecan and a polyacrylamide‐entrapped aggrecan particle assay. Proteolytic activity was measured using a novel, nonglycosylated, reporter substrate assay. Results All forms of ADAMTS‐4 were active to varying degrees in the reporter substrate assay. Digestion of aggrecan in solution digests was apparent in all proteins with the exception of the catalytic domain in isolation (?DTS). Activity towards aggrecan decreased with increasing truncation of the protein. Discussion Removal of the cysteine‐rich‐spacer domain and further C‐terminal truncations decrease the activity of ADAMTS‐4 towards aggrecan, whilst the proteolytic activity remains intact. Cleavages releasing the ancillary domains of ADAMTS' may therefore alter the catalytic activity of these enzymes against proteoglycans and also nonglycosylated polypeptides. More information is required about potential substrates for the processed forms of ADAMTS‐4.     

Keratoconus,allergy, itch,eye‐rubbing and hand‐dominance

Background: One hypothesis for the pathogenesis of keratoconus includes teenage allergy, ocular itch and associated eye‐rubbing. Methods: This study examined the prevalence of these factors for teenage and adult patients. The results for a sample of 53 subjects with bilateral keratoconus were compared with those for a control sample of non‐keratoconus subjects, who also routinely wore RGP contact lenses. The strongest dominant hand and the eye with more advanced keratoconus were also determined, to examine for a relationship between them. Results: The keratoconic sample reported significantly higher levels of allergy, itch and rubbing as teenagers and as adults. However, all distributions were bimodal, consistent with the hypothesis that allergy, itch and rubbing are relevant in the pathogenesis of keratoconus only when the highest levels of these factors are present. For example, a significant relationship between the stronger dominant hand and the more advanced eye was evident only in subjects who reported the most severe rubbing. Conclusions: This finding adds weight to the circumstantial evidence that rubbing contributes to the pathogenesis of keratoconus. Low levels of teenage rubbing by some keratoconic subjects suggest a non‐rubbing pathogenesis and that emphasis on rubbing management is not warranted in these cases. However, high levels of adult rubbing reported by many keratoconic subjects indicate that the standard advice to avoid vigorous and prolonged rubbing is often not effective, even when repeated. There appears to be an indication for the need to improve the management of eye‐rubbing for some patients with keratoconus or at risk of developing this disease.