Reduction of target gene expression by a modified U1 snRNA.

Although the primary function of U1 snRNA is to define the 5' donor site of an intron, it can also block the accumulation of a specific RNA transcript when it binds to a donor sequence within its terminal exon. This work was initiated to investigate if this property of U1 snRNA could be exploited as an effective method for inactivating any target gene. The initial 10-bp segment of U1 snRNA, which is complementary to the 5' donor sequence, was modified to recognize various target mRNAs (chloramphenicol acetyltransferase [CAT], beta-galactosidase, or green fluorescent protein [GFP]). Transient cotransfection of reporter genes and appropriate U1 antitarget vectors resulted in >90% reduction of transgene expression. Numerous sites within the CAT transcript were suitable for targeting. The inhibitory effect of the U1 antitarget vector is directly related to the hybrid formed between the U1 vector and target transcripts and is dependent on an intact 70,000-molecular-weight binding domain within the U1 gene. The effect is long lasting when the target (CAT or GFP) and U1 antitarget construct are inserted into fibroblasts by stable transfection. Clonal cell lines derived from stable transfection with a pOB4GFP target construct and subsequently stably transfected with the U1 anti-GFP construct were selected. The degree to which GFP fluorescence was inhibited by U1 anti-GFP in the various clonal cell lines was assessed by fluorescence-activated cell sorter analysis. RNA analysis demonstrated reduction of the GFP mRNA in the nuclear and cytoplasmic compartment and proper 3' cleavage of the GFP residual transcript. An RNase protection strategy demonstrated that the transfected U1 antitarget RNA level varied between 1 to 8% of the endogenous U1 snRNA level. U1 antitarget vectors were demonstrated to have potential as effective inhibitors of gene expression in intact cells.

The development and evaluation of a computer-assisted teaching programme for intrapartum fetal monitoring.

OBJECTIVE: The development and evaluation of a computer-assisted teaching programme of cardiotocography and acid-base balance. DESIGN: Randomised controlled trial. PARTICIPANTS: One hundred and seventeen midwifery and obstetric staff at Derriford Hospital, Plymouth. METHODS: The obstetricians and midwives were randomly allocated to use the teaching programme, either early or late. The late group (control) used the teaching programme three months after the early group. To assess the effect of the teaching programme, participants were tested on four occasions over eight months by a multiple choice questionnaire. Two questionnaires on ease of use were also completed. MAIN OUTCOME MEASURES: Multiple choice questionnaire scores and opinion questionnaire results. RESULTS: The mean score in the early group improved from 50-8% (test 1, pre-teaching programme) to 70.2% (test 2, post-teaching programme). The mean score in the control group was 50.3% (test 1) and 54.8% (test 2). Knowledge was retained up to seven months. CONCLUSIONS: The teaching programme was effective in improving knowledge of acid-base balance and cardiotocography and can be used by all staff whilst on duty on the labour ward.

Umbilical cord blood gas analysis at the time of delivery.

AIMS: it is now recommended that cord blood acid-base measurement is performed routinely at time of delivery in the UK as a measure of fetal response to labour. However, there remains some uncertainty about the value of this procedure. In this paper our experience of cord blood analysis is described and the literature is reviewed to: (1) provide an overview of the physiological basis of cord blood acid-base assessment; (2) describe the appropriate methodology and identify issues which have contributed to confusion and undermined the value of cord blood sampling; and (3) address the practical issues of cord blood sampling. CONCLUSIONS: cord blood acid-base measurement has a sound physiological basis. It provides objective information which is a useful adjunct to subjective methods of newborn assessment, enables babies at risk of neonatal morbidity to be identified, can be helpful in litigation cases and is a prerequisite for clinical audit. However, to be of benefit the information must be correct and correctly interpreted.

The QSHP proposal: 10 questions most often asked.

Qualified Student Health Plans (QSHPs) have gained increasing support since the authors introduced the proposal to the field of college health in 1994. This article answers 10 of the most frequently asked questions about QSHPs and summarizes changes in the proposal that have resulted from discussions among college health professionals throughout the country. The QSHP proposal, which has been endorsed by the board of directors of the American College Health Association, offers the following benefits for the college health field and for students: It is founded on the traditional prepaid college health model and assures that control remains with higher education; participation is voluntary for employers and for institutions of higher education; no new tax funding is required; QSHPs preclude state interference and provide employers with incentives to contribute to the cost of student healthcare; both the cost of student health insurance and duplication of coverage are reduced and benefits are shifted to where the students live; the problem of uninsured and underinsured students is eliminated at participating institutions. With incremental healthcare reform continuing at the state level, the time to move forward with the proposal is viewed as ideal.

A multicentre comparative study of 17 experts and an intelligent computer system for managing labour using the cardiotocogram.

OBJECTIVES: To investigate 1. whether an intelligent computer system could obtain a performance in labour management comparable with experts when using cardiotocograms (CTGs), patient information, and fetal blood sampling and 2. whether experts could be consistent and agree in their management of labour. SUBJECTS: An intelligent computer system and 17 clinicians experienced in fetal monitoring from 16 centres in the UK. DESIGN: Fifty cases with complete intrapartum CTGs and clinical data were reviewed by each expert and the system independently on two occasions, at least one month apart. Each CTG was scored in 15 min segments according to a protocol and estimates of the cervical dilatation and fetal scalp blood pH were given when requested. MAIN OUTCOME MEASURES: Consistency and agreement in the recorded scores, agreement and timing of cases recommended for caesarean sections, fetal blood sampling rates, intervention in cases with poor outcome and intervention in cases with good clinical outcome. RESULTS: The system: 1. Agreed with experts well and significantly better than chance (67.33%, kappa = 0.31, P << 0.001). 2. Was highly consistent (99.16%, kappa = 0.98, P << 0.001) when used by two operators independently. 3. Recommended no unnecessary intervention in cases with normal delivery and good condition (cord artery pH > 7.15, vein pH > 7.20, 5 min Apgar > or = 9 and no resuscitation). This was better than all but two of the experts. 4. Recommended delivery by caesarean section in 11 cases; at least 15 of the 17 experts in each review also recommended caesarean section delivery in these cases. The majority did so within 15 min of the system and two-thirds did so within 30 min. 5. Identified as many of the birth asphyxiated cases (cord arterial pH < 7.05 and BDecf > or = 12, and Apgar score at 5 min < or = 7 with neonatal morbidity) as the majority of experts and one more than was acted upon clinically. The experts were found to be consistent and to agree. There was good agreement in the cases and the timing of caesarean section recommendations. The majority of experts did not recommend operative intervention in cases which had a normal delivery and good outcome, but did recommend operative interventions in 10 of 12 cases delivered with cord arterial pH < 7.05. However, in one of the cases delivered with birth asphyxia, 14 of the 17 experts and the system failed to recommend intervention. CONCLUSIONS: The system's performance was found to be indistinguishable from the experts' in the 50 cases examined, but it was more consistent. This demonstrates the potential for an intelligent computer system to improve the interpretation of the CTG and decrease intervention. Furthermore, the good performance of most experts in this study demonstrates the potential effectiveness of the CTG and raises important questions regarding why the CTG has fallen short of expectations in current practice.

Delegation to assistive personnel by school nurses--one state's experience.

Budget cuts to education, reduction of school health services and an increase in the nursing needs of school-age children have placed pressure on school nurses to delegate nursing procedures to assistive personnel. This paper reports on a study of school nurses in a Midwestern state. Nurses reported refusing to delegate, delegating against their better judgment, delegating without knowledge of the assistant's qualifications or without providing orientation or training, and delegating because they were told to by someone else. The study identified factors considered by nurses when deciding to delegate a procedure, educational preparation for delegation, and problems experienced with delegation.

Student healthcare delivery and financing programs: adapting to healthcare reform.

College health professionals want to assure the unique healthcare and health education needs of college students will continue to be met under national and state healthcare reform. This may be an "all or nothing" proposition. Either colleges and universities will have exclusive control of healthcare delivery for the college student population or else college health will not be a major force in healthcare reform. If college health is to play a meaningful role in future government-controlled health insurance programs, it must first demonstrate that current health services and insurance financing programs meet minimum quality standards. This proposal calls for expanding existing federal laws to create qualified student health plans and integrating the college health model into a reform package based on employer-sponsored health insurance. The concept of qualified student health plans allows for a high degree of flexibility that can be integrated into the majority of state and federal healthcare reform proposals, including the plan proposed by President Clinton, that are not based on a single-payer system. Ultimately, the authors suggest, their proposed plan would eliminate the current situation, in which large numbers of college students are uninsured or underinsured.

Scar rupture in labour after previous lower uterine segment caesarean section: the role of uterine activity measurement.

A series of 12 trials of scar associated with scar rupture is reviewed. Uterine activity patterns were assessable in 10 of them. Clinical features and characteristics of the intrauterine pressure waveform and uterine activity are discussed in relation to the integrity of the scar.

Comparison of immunologic and enzymatic assay of prostatic acid phosphatase for follow-up and assessment of clinical status of stage D prostate cancer.

Prostatic acid phosphatase (PAP) was measured in 70 patients with stage D prostate cancer under different modalities of treatment. PAP was determined by radioimmunoassay (RIA), counter immunoelectrophoresis (CIEP), and enzymatic method using alpha-naphthyl phosphate to compare the usefulness of the three methods in follow-up and assessing the clinical status of stage D prostate cancer. In the regressive state (29 patients), RIA and enzymatic methods correlated well; both gave 17% of abnormal results with a mean value of 4.7 +/- 4.6 and 3 +/- 1.7. Also, in the progressive state (17 patients) the two methods showed similar percentages of abnormal results with a mean value of 40 +/- 38 and 19 +/- 17 for RIA and enzymatic method, respectively. There was greater variability in the stable group owing to the difference in the tumour load. Again the two methods correlated well regarding their diagnostic sensitivity and specificity as a parameter for assessing the clinical response. CIEP, used as a qualitative method, showed more positive than negative results and did not correlate with the clinical state. We feel that the conventional enzymatic method is adequate for follow-up and assessing clinical state of stage D prostate cancer.

Treatment of sequential bilateral germ cell tumors of the testis following interval retroperitoneal lymph node dissection.

Modification in lymphatic drainage following retroperitoneal lymph node dissection, such as a collateral circulation or lymph node and lymphatic vessel regeneration, was observed in 2 patients in whom a second tumor developed in the remaining testicle. Such alterations of the lymphatic system are difficult to evaluate for the possible presence of metastatic disease. The presence of extensive collateral circulation rules out lymph node dissection or radiation therapy as an appropriate treatment in these patients. A short course of systemic chemotherapy, regardless of the histological type of the second malignancy, seems to be the safest adjunctive treatment in such cases.

The addition of chemotherapy to hormonal therapy for treatment of patients with metastatic carcinoma of the prostate.

Patients with advanced prostate carcinoma that had been stabilized by orchiectomy (ORCH) or hormone therapy for at least 3 months, were randomized to either diethylstilbestrol (DES) alone or DES plus Cytoxan or DES plus Emcyt. A total of 188 patients were randomized between July, 1976 and February, 1982 of which 161 were evaluable for objective response to treatment. Objective response rates, response duration, or survival experiences were not demonstrably different between treatment arms, either for all patients or within good or poor prognosis groups determined by initial pain or acid phosphatase level. Subjective improvements in performance status were small for each treatment. Pain relief was somewhat greater in the chemotherapy-hormone combinations than in the DES/ORCH, but the advantage was not statistically significant. Side effects were primarily nausea and vomiting and leukopenia, mostly in the DES + Cytoxan arm. The duration of stabilization prior to entry did not influence response overall, although there were opposing trends within each of the two chemotherapy arms. The premise for combining antitumor agents with hormones before hormone failure is still felt to be a more logical approach than waiting for the ultimate hormone failure, and a combination of hormones plus two antitumor agents is being evaluated in a subsequent ongoing trial where a more rigid design limits the duration of the preentry period of hormone stabilization.

Comparison of estramustine phosphate, methotrexate and cis-platinum in patients with advanced, hormone refractory prostate cancer.

In this clinical trial of men with advanced prostatic cancer no longer responsive to hormone therapy 189 were randomized to receive estramustine phosphate, methotrexate or cis-platinum. Response evaluations were done in 158 cases. Objective response rates (complete, partial or stabilization of disease) were 34 per cent for estramustine phosphate, 36 per cent for cis-platinum and 41 per cent for methotrexate. Subjective parameters indicated a substantial advantage for pain improvement with methotrexate or cis-platinum over estramustine phosphate. Probabilities of continued response indicated some advantage for methotrexate and median response durations at this time were twice as long for methotrexate (32 weeks) as for cis-platinum (16 weeks), with estramustine phosphate intermediate (23 weeks). Survival rates for the original treatment randomization groups were not different at this time. Side effects of estramustine phosphate consisted primarily of nausea and vomiting and/or anorexia but to a lesser extent than with cis-platinum. These effects were somewhat less for methotrexate, for which the major side effects were stomatitis and leukopenia, as well as hepatic toxicity reflected by elevated serum glutamic oxaloacetic transaminase levels. Other side effects of cis-platinum were less than for methotrexate (no stomatitis), except for signs of renal toxicity (elevations in blood urea nitrogen and serum creatinine), which were greater. Methotrexate had a relatively high level of activity against metastatic, progressive, hormone nonresponsive prostatic cancer, with side effects that were substantial but manageable.

The treatment of Stage III nonseminomatous testicular tumors. Roswell Park Memorial Institute results (1970-1979).

A review of 92 patients with Stage III nonseminomatous tumors treated at Roswell Park Memorial Institute between 1970-1979 was undertaken to verify changes in concepts as related to multiple agent chemotherapy and cytoreductive surgery. Each patient had a minimal follow-up of 18 months. Fifty-three patients were seen before 1975. Eighteen had metastasis to the lungs only. These were treated with a variety of single chemotherapeutic agents and cytoreductive surgery. The survival of this group was 38%. Among 35 patients with lung and visceral involvement seen at the same time, only one patient is alive. Thirty-nine patients were seen after 1975 and treated with multi-drug chemotherapy and cytoreductive surgery. The current survival rate of 23 patients with lung metastasis only is 69%. Among 16 patients with lung and visceral involvement, the present survival rate is 31%. This report confirms the effectiveness of multi-drug therapy in conjunction with cytoreductive surgery in the treatment of disseminated testicular tumors.

Filling of the obturator nodes in pedal lymphangiography: fact or fiction.

The value of pedal lymphangiography as a staging procedure for carcinoma of the prostate and the bladder continues to be debated because of the fact that the obturator lymph nodes apparently are not visualized during this diagnostic technique. This study involves 25 patients who have undergone pedal lymphangiography followed by fine needle aspiration. Pelvic lymph node dissection was performed and pelvic x-rays were taken once the external iliac nodes had been excised, leaving the obturator nodes behind. The next step was the removal of the remaining nodes in the obturator fossa, after which another x-ray was obtained. With this technique we could prove whether these obturator lymph nodes opacified on routine pelvic x-ray. In a review of the radiography consistent filling of the obturator nodal chain was noted in all of the cases, as well as the consistent prediction of the location of these lymph nodes before fine needle aspiration.

High-dose cytoreductive therapy with autologous bone marrow transplantation in advanced malignancies.

Twenty-one patients with advanced malignancies received high-dose chemotherapy and/or radiotherapy followed by autologous bone marrow infusion. Eighteen patients (85.7%) had fever greater than or equal to 100 degrees F for a median of 6 days; 14 of these patients required broad-spectrum antibiotics for a median of 13 days. Nineteen patients (90.5%) had a granulocyte count less than 500/mm3 for a median of 11 days. Thrombocytopenia (platelet count less than 50,000/mm3) was observed in 18 patients (85.7%) for a median of 14 days. Mucositis and diarrhea were not common, occurring in six (28.6%) and seven (33.3%) patients, respectively. Of the 21 patients studied, 16 were evaluable for tumor response; there were four complete responses and four partial responses, and two patients who showed no change for variable times. Two patients have unmaintained remissions for greater than 2 years. Our response rate (complete plus partial) is 50%. Our study shows that high-dose cytoreductive therapy can be given with moderate toxicity when combined with autologous bone marrow infusion. Because responses in this group of patients are generally of short duration, we believe that patients with advanced malignancies who have had less exposure to therapy or who have a high likelihood of disease recurrence should be considered for high-dose cytoreductive therapy with autologous transplantation.

Changing concepts in the treatment of advanced seminomatous tumors.

The treatment of patients with advanced seminomatous tumors represents a challenge for the medical and urological oncologists. In contrast to the reported excellent results of radiotherapy in the treatment of patients with clinical stages I and II disease the survival rate for those with advanced seminomatous disease is only around 40 per cent. Six consecutive patients with metastatic seminomas were treated with systemic chemotherapy and all achieved a complete response. The combination of vincristine, cis-platinum, bleomycin and prednisone was used in 4 patients and an excellent tolerance was observed even in patients with previous radiation therapy. Two other patients who had had radiation therapy to the mediastinum were treated with VP-16 and cis-platinum and again a complete response was observed. However, a significant potentiation of myelosuppressive effect of VP-16 was observed in the previously radiated patients. In view of the observed sensitivity of seminomatous tumors it appears that all patients who present initially with metastatic or bulky retroperitoneal disease should be treated with multiple drug chemotherapy and cytoreductive surgery similar to the treatment modalities used for nonseminomatous germ cell tumors. Radiation therapy for patients with this stage of the disease seems to be not only ineffective but also may compromise the benefits of aggressive chemotherapy.

A comparison of estramustine phosphate versus cis-platinum alone versus estramustine phosphate plus cis-platinum in patients with advanced hormone refractory prostate cancer who had had extensive irradiation to the pelvis or lumbosacral area.

Single and combination chemotherapy was compared in a clinical trial for men with advanced, metastatic prostate cancer who had received prior pelvic irradiation and had had progression of disease despite hormonal therapy. The 149 patients were randomized to receive estramustine phosphate or cis-platinum alone or in combination. Of the 149 patients 25 (17 per cent) were excluded from the study but 124 were evaluated for response and survival. Entry variables were distributed similarly among patients in each treatment arm. There were no complete or partial responders but there were nearly twice as many patients whose disease was stabilized (33 per cent) on the combination regimen compared to estramustine phosphate (18 per cent) and about a third more than for cis-platinum (21 per cent). Analysis of survival revealed some advantage for patients on combination therapy. Major toxicities for all treatments were nausea and vomiting (62 to 88 per cent) and accompanying anorexia (72 to 95 per cent). Azotemia developed in 45 per cent of the patients receiving combination therapy. In addition an elevation in serum creatinine occurred in 22 per cent of the patients receiving combination therapy and in 17 per cent of those receiving cis-platinum alone. Myelosuppression occurred infrequently.

Intermittent hemodialysis therapy in the cancer patient.

The occurrence of renal failure in the cancer patient presents a series of medical, technical, philosophical, and logistic problems that are rarely found in other patient population groups. In a review of 70 cancer patients requiring intermittent hemodialysis therapy at Roswell Park Memorial Institute over the last 6 years for acute renal failure, 46 (65.7%) of the patients were salvaged from renal failure to allow the use of further therapeutic modalities for their neoplastic disease. Patient survival after successful intermittent hemodialysis therapy was comparable to that expected in the nonazotemic cancer patients submitted to similar anticancer therapy. Hemodialysis should, therefore, be considered and offered in selected cancer patients in acute renal failure.