Isoform specific changes in calcium transporters in colon and breast cancer cell lines

Calcium transporters play vital roles in the transport of calcium ions across cells of the mammary gland and the intestine. One such transporter is the plasma membrane Ca2+-ATPase (PMCA), of which there are 4 different genes (PMCA1-4). In these studies we investigated the hypothesis that the expression of PMCA is altered in HT-29 colon cancer cells during sodium butyrate and post-confluence mediated differentiation. We also investigated if PMCA expression is altered in breast cancer cell lines in an isofrom specific manner. Our results indicate isoform specific changes in PMCA mRNA and protein levels in HT-29 cells during differentiation, using real time RT-PCR and western blotting, respectively. We also observed pronounced alterations in the mRNA levels of the PMCA isoform linked to lactation (PMCA2) in a bank of breast cancer cell lines compared to normal cell lines. Changes in other isoforms were less pronounced. To further study the role of specific calcium transporters we have optimised conditions for the reverse transfection of MCF-7 breast cancer cells using NeoFX (Ambion). Using real time RT-PCR we have confirmed gene knockdown for specific isoforms and have studied the time course of knockdown over 96 hours. We see approximately 68 % inhibition at 24 hours increasing to 84 % 96 hours post-reverse transfection. Our studies suggest that the expression of specific calcium transporter isoforms can be significantly altered in cancer cell lines and that isoform specific inhibition of calcium transporters is possible using reverse transfection of siRNA

Uncoupled bone turnover in cystic fibrosis: bone markers and the PTH-vitamin D axis

No abstract

Evaluation of a pilot telemedicine network for accident and emergency work

A pilot accident and emergency. teleconsulting service was established in Scotland. It was based at the accident and emergency department of the main hospital in Aberdeen. There were three peripheral sites in rural Grampian (Peterhead, Turriff and Huntly) and one in the Shetland Isles. The videoconferencing equipment used was connected by ISDN at 384 kbit/s. During the 15 months of the study, 1998 videoconference calls were made, of which 402 (20%) calls were made to the accident and emergency department for clinical consultations. The majority of the clinical calls (95%) were made between 09:00 and 17:00, and more than 90% were completed within 20 min. During the majority of calls (87%) one or more X-ray images were transmitted. The majority of patients (89%) received treatment without transportation to the main centre in Aberdeen. The present study demonstrated that accident and emergency teleconsultations can be technically reliable, effective in reducing the number of patient transfers and acceptable to the referring clinicians. As a result, approximately pound1.5 million has been made available by the government to develop a national system for Scotland.

Design and implementation of an automatic message-routing system for low-cost telemedicine

Email has been used for some years as a low-cost telemedicine medium to provide support for developing countries. However, all operations have been relatively small scale and fairly labour intensive to administer. A scalable, automatic message-routing system was constructed which automates many of the tasks. During a four-month study period in 2002, 485 messages were processed automatically. There were 31 referrals from eight hospitals in three countries. These referrals were handled by 25 volunteer specialists from a panel of 42. Two system operators, located 10 time zones apart, managed the system. The median time from receipt of a new referral to its allocation to a specialist was 1.0 days (interquartile range 0.7-2.4). The median interval between allocation and first reply was 0.7 days (interquartile range 0.3-2.3). Automatic message handling solves many of the problems of manual email telemedicine systems and represents a potentially scalable way of doing low-cost telemedicine in the developing world.

The importance of setting and evaluating standards of telemedicine training

The importance of appropriate training in the use of videoconferencing equipment for clinical purposes is often underestimated when telemedicine projects are established. We developed a user training programme which was delivered via videoconferencing to a group of 130 nurses. Training was delivered on a one-to-one basis. A questionnaire was developed to evaluate user satisfaction and the effectiveness of training. One hundred and two fully completed questionnaires were returned (a 79% response rate). High levels of satisfaction were obtained but the level of user competence reached 100% only when training was supported by a training manual and at least weekly practice. Before establishing a telemedicine service, the following steps appear to be important: identify the required training competencies; deliver a 'hands on' training programme based on the required training competencies; back up the training programme with an instruction booklet; ensure that trainees have at least weekly practice; measure the level of user competence.