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Biochemiluminescence and biomedical applications.

Although used for analytical purposes for more than 40 years it is only recently that biochemiluminescence (BCL) has found widespread acceptance. Methods employing BCL reactions now play an important role in biomedical research and laboratory medicine. The main attractions for the assay technology include exquisite sensitivity (attomole-zeptomole), high selectivity, speed and simplicity. In biomedical research, the most important applications of BCL are: (1) to estimate microbial numbers and to assess cellular states (e.g., after exposure to antibiotic or cytotoxic agents) and in reporter gene studies (firefly luciferase gene); (2) NAD(P)H involved in redox/dehydrogenase studies using Vibrio luciferase complex; (3) BCL labels and CL detection of enzyme labels in immunoassays are the most widespread routine application for this technology. BCL enzyme immunoassays represent the most active area of development, e.g., enhanced BCL method for peroxidase and BCL assays for alkaline phosphatase labels using adamantyl 1,2-dioxetane.

Bacteria↗

Efficacy of low-dose desferrioxamine for the estimation of aluminium overload in haemodialysis patients.

In haemodialysis patients the desferrioxamine (DFO) test is an accepted method for estimating the body content of aluminium. However, toxic side effects have been frequently reported with the high-dose (30 or 40 mg/kg) DFO tests. To study if a low-dose (500 mg) DFO test was also useful and free of side effects, we compared it with a high-dose DFO test in 22 patients on regular haemodialysis. During the first two hours of dialysis, 500 mg DFO in 100 ml 0.9% NaCl were administered intravenously. Before dialysis and after 48 hours aluminium concentration was determined in serum. In 14 patients the low-dose DFO test was considered positive. In 11 of those 14 patients the high-dose (30 mg/kg) DFO test was positive too. Analysis of the results showed a significant correlation (p < 0.05) between the outcome of the two DFO tests in the same patient. Compared to the high-dose DFO test, the low-dose test revealed a sensitivity of 79% and a specificity of 63%. In contrast with the high-dose DFO test might be a reliable and safe method for the estimation of the body content of aluminium in patients on regular haemodialysis.

Adult↗

Issues in protection from galactic cosmic rays.

Radiation risks to astronauts depend on the microscopic fluctuations of energy absorption events in specific tissues. These fluctuations depend not only on the space environment but also on the modifications of that environment by the shielding provided by structures surrounding the astronauts and the attenuation characteristics of the astronaut's body. The effects of attenuation within the shield and body depends on the tissue biological response to these microscopic fluctuations. In the absence of an accepted method for estimating astronaut risk, we examined the attenuation characteristics using conventional linear energy transfer (LET)-dependent quality factors (as one means of representing relative biological effectiveness, RBE) and a track-structure repair model to fit cell transformation (and inactivation) data in the C3H10 T1/2 mouse cell system obtained for various ion beams. Although the usual aluminum spacecraft shield is effective in reducing dose equivalent with increasing shield thickness, cell transformation rates are increased for thin aluminum shields. Clearly, the exact nature of the biological response to LET and track width is critical to evaluation of biological protection factors provided by a shield design. A significant fraction of biological injury results from the LET region above 100 keV/mu m. Uncertainty in nuclear cross-sections results in a factor of 2-3 in the transmitted LET spectrum beyond depths of 15 g/cm2, but even greater uncertainty is due to the combined effects of uncertainty in biological response and nuclear parameters. Clearly, these uncertainties must be reduced before the shield design can be finalised.

Animals↗

Assessment of the value and pattern of use of a target controlled propofol infusion system.

Thirty one anaesthetists were supplied with a pharmacokinetic based target controlled propofol infusion system for evaluation. Twenty seven of 30 replies to a questionnaire sent to them indicated that the system had changed their use of propofol for maintenance of anaesthesia. The main reasons were greater ease of use and more confidence regarding the predictability of anaesthetic effects compared with manually controlled infusion. Data obtained from 770 patients anaesthetised with the system were analysed. The median maximum target concentration selected was 6.6 micrograms/ml. Younger patients (18-35 yr) required significantly greater target concentrations than older patients (65-80 yr). The mean time during which the system was in maintenance mode, when the predicted blood concentration of propofol was held constant for at least one minute, was 26.1 minutes. The median number of alterations in propofol concentration was 6. The target controlled infusion system provided an inexpensive and acceptable method of delivering intravenous anaesthesia.

Adolescent↗

Unilateral inferior temporal lobectomy with hippocampectomy for relief of incisural herniation.

1. The syndrome of incisural hippocampal herniation, accompanied by unilateral and then bilateral fixed pupils, deepening coma, respiratory irregularity (Cheyne-Stokes), contralateral hemiplegia, ipsilateral decerebrate posturing rising blood pressure, and finally renal shutdown, indicates a terminal state that requires immediate relief. 2. Accepted methods of relief by brain shrinkage, dexamethasone, subtemporal decompression, anterior temporal lobectomy, cutting of the tentorium, and even massive uncapping of the skull, have failed in the author's experience to reverse the process when once established. 3. Presentation is made of a rapid and relatively simple inferior horizontal temporal lobectomy, including the hippocampal gyrus, so as to relieve peduncle compression, blockage of cerebrospinal fluid circulation, midbrain haemorrhage, and infarction in the posterior cerebral artery distribution area. 4. Reports are given of 15 cases out of 30 cases in which operation proved lifesaving, with restoration of normal function in 13. 5. Those patients who died did so in an average of five days, and generally showed either no herniation or continuing evidence of tumour or internal bleeding. 6. Very young patients particularly have an excellent chance of survival, which is seven times better than that of adults in having good to excellent results without operation.

Adolescent↗

On-line computer estimation of carbon dioxide response curves.

Anesthesiologists are concerned with the effect of various anesthetics on a patient's central nervous ventilatory control. The most widely accepted method of determining the effect of a drug is to compare carbon dioxide response curves (delta VE/delta PETCO2, where VE = minute ventilation [in L/min] and PETCO2 = end-tidal carbon dioxide [in mm Hg]) measured before and after administration of the drug. Additional information concerning neuromechanical control can be obtained by also including a measure of the airway occlusion pressure (generally measured 100 ms after occlusion, i.e., P100). To facilitate these measurements we have developed a portable, computer-controlled data acquisition system. It includes an Apple II+ computer and measures VE, PETCO2, and P100. Each subject rebreathes exhaled carbon dioxide through a two-way breathing valve attached to a 9-liter reservoir, which is initially filled with 5% carbon dioxide and balance oxygen. Exhaled carbon dioxide concentrations are measured with an infrared medical gas analyzer on samples taken through a catheter connected at the mouthpiece. The exhaled flow is measured with a pneumotachograph in conjunction with a differential pressure transducer, and P100 is determined with a Validyne MP45 pressure transducer.

Airway Resistance↗

Five-year evaluation of safety, efficacy and acceptability of Norplant implants in Nepal.

This paper presents findings based on a five-year, noncomparative study of Norplant contraceptive subdermal implants in Nepal. The study was designed to evaluate the contraceptive safety, efficacy, and overall acceptability of Norplant. Four hundred and seven women enrolled in the clinical trial, which began in 1985, at two study sites, located in Patan and Kathmandu. Follow-up visits were scheduled at 1, 3 and 6 months after Norplant insertion and every six months thereafter until removal or at the end of five years. Although five pregnancies were reported during the study, only two women (one from each center) were diagnosed as becoming pregnant while using Norplant. The pooled gross cumulative life-table pregnancy rate was 0.6 per 100 women at the end of five years. The pooled cumulative continuation rate was 62 per 100 women at the end of five years. The three most frequently reported reasons for discontinuation were menstrual problems, personal reasons, and medical reasons. Of the 125 women who completed a five-year user satisfaction questionnaire, the majority of the women (86%) planned to continue using contraception after study completion. Of these women, almost one half said they planned to use a second Norplant set. The findings suggest that the Norplant system is a safe, effective, and acceptable method of contraception among Nepalese women.

Adolescent↗

The frameless GyneFix intrauterine implant: a major improvement in efficacy, expulsion and tolerance.

The CuFix (GyneFix), conceived in 1985, was developed to minimize three major problems frequently associated with discontinuation of IUD use: expulsion, bleeding and pain. Since the initial clinical investigations, over 10,000 woman years of experience and up to 8 years of follow-up in international multicenter, non-comparative and comparative clinical trials, including a large proportion of nulligravid/nulliparous women, have been collected. Based on new clinical information about the GyneFix from a long-term multicenter clinical trial, conducted in young nulligravid/nulliparous and parous women, the importance of this new contraceptive is discussed. The following conclusions were reached: The unique design characteristics of the GyneFix (frameless, flexible and fixed to the fundus of the uterus) have resulted in optimal tolerance and almost complete absence of expulsion. The result is enhanced effectiveness (comparable to OCs and male/female sterilization) and a high rate of continued use. The GyneFix reduces the IUD failure rate to a minimum and is, therefore, a welcome reversible alternative to OCs and female surgical contraception. Framelessness and flexibility explain the absence of side-effects and adverse events caused by dimensional incompatibility between the frame of conventional IUDs and the uterine cavity and may also explain the absence of PID and ectopic pregnancies in any of the clinical studies. The GyneFix is a promising new, highly effective and safe, contraceptive option for parous women and an equally effective and well-accepted method for nulliparous women.

Adolescent↗

Hirschsprung's disease in adolescents and adults.

Hirschsprung's disease in the adolescent and adult is a rare and often misdiagnosed cause of lifelong refractory constipation. Two adolescent and three adult patients with Hirschsprung's disease treated between 1973 and 1987 at the University of Michigan Medical Center are reported. Each patient presented with chronic constipation requiring enemas, cathartics, and multiple hospital admissions for management. Diagnosis in each case was made with barium enema and full-thickness rectal biopsy. Four patients underwent endorectal pull-through procedures, all with good long-term results. The fifth patient, initially treated with a Duhamel retrorectal pull-through procedure, required reoperation for constipation secondary to a retained rectal septum. Review of 199 cases of adult Hirschsprung's disease enables comparison of the various operative procedures for this disorder with respect to postoperative complications and functional outcomes. Anorectal myectomy with low anterior resection, the Duhamel-Martin procedure, and the Soave endorectal pull-through procedure are the most acceptable methods for surgical management.

Adolescent↗

Comparison of heart rate variability spectra using generic relationships of their input signals.

Spectral analysis of heart rate variability (HRV) is an accepted method for assessment of cardiac autonomic function and its relationship to numerous disorders and diseases. Various non-parametric methods for HRV estimation have been developed. The spectrum of counts, the instantaneous heart rate spectrum and the interval spectrum are mostly practised. Although extensive literature on their respective properties is available, there seems to be a need for a more complete comparison, eventually resulting in recommendations for applicability. The methods for HRV spectral analysis use their specific transforms of the primary R-R interval series into input signals for spectral computation. This is, in fact, the reason for obtaining different spectra. A basis for comparison is established, revealing the generic relationships of these HRV input signals. It allows for a more systematic evaluation and for further development of the considered methods. The results with simulated and real HRV data show better performance by the spectrum of counts and by a proposed instantaneous heart rate spectrum, obtained using a cubic spline interpolated input signal. The modulation depth of the primary signal can influence the accuracy of the HRV analysis methods.

Electrocardiography↗

Expectations of recipient couples awaiting an anonymous oocyte donor match.

PURPOSE: Our purpose was to survey recipients in an ovum donation program and report on their expectations while waiting for their potential donor recipient match. METHODS: Accepted or rejected anonymous ovum donor matches (n = 80) from January 1996 to May 1997 were evaluated. Patients generated a "wish list" of desired traits and physical characteristics. From an approved donor pool of medically and psychologically screened women, candidates were drawn upon as potential matches and presented to the potential recipient, who decided whether to accept the donor. Reasons for accepting or rejecting the donor were tallied and were compared to the patient's wish list. RESULTS: Medical history and race were ranked by 33 and 23% of recipient couples as the two most important characteristics, while 74 and 54% stated that these were among the three most important factors in a potential donor compared with other traits. Fifty-seven (71%) recipients accepted, while 23 (29%) rejected, the first donor presented to them. Eleven were subsequently given a second choice within 6 months, with 10 (91%) accepting the next presented match. Recipients waiting for a donor were just as likely to accept or reject a potential candidate whether waiting < 3 months (33%; 15/46), 3-6 months (25%; 4/16), or > 6 months (22%; 4/18) (P > 0.05; NS). In all but five recipients, the reason for rejection was consistent with their top three priorities reported in their wish list. CONCLUSIONS: Phenotypic, ethnic, educational, and other interests are important in the selection of an ovum donor. Recipients are proactive in their decision process, making educated and well considered decisions in spite of the limited pool and the extended time frame in waiting for an appropriate ovum donor.

Attitude↗

Inhibitory effect of an oxygenated cholesterol on the induction and progression of DMBA-induced mammary carcinomas in the rat.

In vivo studies on the effect of two stereoisomeric 7,22-dihydroxycholesterols on tumor development were conducted in the Charles Huggins animal cancer model (DMBA-induced mammary cancer in the Sprague-Dawley female rat). Three groups of DMBA-treated animals were fed a 9:1 mixture of (22R)-cholest-5-ene-3 beta,7 beta,22-triol and (22R)-cholest-5-ene-3 beta,7 alpha,22-triol in the drinking water in a calculated dose of 250 micrograms per animal per day. One group (A) received the sterols throughout the experimental period of 35 weeks, another group (B) during the first 12 weeks only, and a third group (C) only during weeks 13 through 35 after DMBA injection. Tumor rates and tumor yields were calculated, and statistical assessment by accepted methods demonstrated a very significant inhibitory effect on tumor development in Groups A and B, as compared with Group C. The results indicate a growth-inhibitory effect during the induction period of carcinoma development. The influence on neoplastic growth of (22R)-cholest-5-ene-3 beta,7 alpha,22-triol, (22R)-cholest-5-ene-3 beta,7 beta,22-triol, and (22R)-cholest-5-ene-3 beta,22-diol-7-one was examined in suspension cultures of Ehrlich ascites tumor cells. The 7 alpha-hydroxy compound proved to be ineffective, whereas the latter two substances displayed a strong cytotoxic effect.

9,10-Dimethyl-1,2-benzanthracene↗

TPA (12-O-tetradecanoyl-phorbol-13-acetate) as a carcinogen for mouse skin. A positive dose-response relationship.

In order to study a possible dose/response relationship in the tumorigenic effect of 12-O-tetradecanoyl-phorbol-13-acetate (TPA), groups of hairless mice were treated topically on the back skin with different doses and treatment schedules of TPA in acetone. A group of 104 mice (56 M/48 F) received a single application of 20 nmol TPA; 80 mice (32 M/48 F) received 2 X 20 nmol TPA at 3 day intervals; 95 mice (48 M/47 F) received five applications; and 32 mice (16 M/16 F) received 50 applications of 20 nmol TPA twice weekly. Two groups of 32 mice (16 M/16 F) were painted topically on the back skin twice weekly with doses of 17 and 10 nmol TPA, respectively, in each application for 60 weeks or until the animals died. All applications were given in 0.1 ml reagent grade acetone. A control group was treated twice weekly with acetone alone for 60 weeks. Long-term application of 17 nmol TPA was toxic to the animals, and only 20% survived 10 months. Doses of 10 nmol TPA twice weekly were less toxic, and 80% of the animals survived 14 months. The other doses were fairly non-toxic. The results were assessed statistically by accepted methods for tumor rates and yields, respectively. There was a significant tumor incidence after the higher doses of TPA and a clear dose-response relationship. This further strengthens the evidence that TPA is a complete carcinogen, and not merely a promoter.

Animals↗

Differential renal function in the prediction of recovery in adult obstructed kidneys after pyeloplasty.

AIM: Pyeloplasty is a widely accepted method for the treatment of ureteropelvic junction obstruction (UPJO). Surgery has long been thought to affect postoperative renal function. However, controversies still exist on the functional studies that can be used to indicate which renal units will benefit from surgery. In this study, the correlation between differential renal function (DRF) and other preoperative parameters was examined to determine which parameter more accurately predicts recovery of renal function in adult obstructed kidneys. MATERIALS & METHODS: In this study, the authors evaluated 32 patients with UPJO. In all patients, standart diuretic Tc-99m DTPA renal scans were performed preoperatively and 6 months after surgery. Patients were divided into two groups according to the preoperative DRF (> or = 30%, n = 22, group I and < 30%, n = 10, group II). Second type classification was made according to the postoperative DRF improvement as improved (group A, n = 13) and not improved (group B, n = 19). These groups were then compared regard to variables which were defined as symptoms, age at operation and ultrasonographic findings. We also evalulated whether preoperative parenchymal function is important to predict improvement in drainage half-time (T1/2). RESULTS: While preoperative drainage half-time was 39.6 +/- 15.9 minutes, postoperative half-time decreased to 16.9 +/- 6.8 minutes (p < 0.001). The mean DRF did not improved significantly after surgery compared with preoperative values (32.03 +/- 9.42% versus 36.16 +/- 9.60%). When comparing the patients with preoperative DRF > or = 30% (group I, DRF 38 +/- 0.8%) to those who had an initial DRF below 30% (group II, DRF 22.8 +/- 5.2%), postoperative DRF was 41.22 +/- 5.72 in group I and 25.00 +/- 6.22 in group II. The difference was significant (p < 0.01). The patients in group I and II showed improvement in 50% and 20%, respectively. Age, clinical presentation and ultrasonographic findings did not affect functional outcome after pyeloplasty. We could not find any correlation between preoperative DRF and the degree of improvement in T1/2. CONCLUSION: Renal function improves after pyeloplasty with regard to the initial level of split renal function in adult obstructed kidneys. Improvement may not be observed especially in patients with DRF less than 30%.

Adult↗

Comparative trial of the effect of ranitidine and cimetidine on gastric secretion in fasting patients at induction of anaesthesia.

A comparative trial of the H2-receptor antagonists, cimetidine and ranitidine, on gastric pH and volume, was conducted in 168 healthy patients coming to elective surgery. The drugs were administered in random fashion either intravenously (ranitidine 50 mg or 100 mg, cimetidine 300 mg or placebo) or orally (ranitidine 150 mg, cimetidine 300 mg or placebo). The patients received the drugs or placebo 45 minutes to five hours before operation. After induction of anaesthesia, a nasogastric tube was passed and the stomach contents were aspirated. The volume and pH were measured. Those patients receiving ranitidine 50 or 100 mg or cimetidine 300 mg intravenously had statistically significantly higher gastric pH compared to those receiving placebo, but up to eight percent of patients has a pH less than 2.5. Oral administration of cimetidine 300 mg or ranitidine 150 mg were also superior when compared to placebo. However, 25 per cent of the patients receiving oral cimetidine had a pH less than 2.5; cimetidine orally was statistically significantly inferior to ranitidine 100 mg given intravenously. We conclude that the intravenous use of either ranitidine or cimetidine is an acceptable method to decrease the acidity of gastric contents before induction of anaesthesia. Orally, ranitidine appears to be a better choice than cimetidine in the doses studied. Both ranitidine and cimetidine need to be given at least 45 minutes before induction of anaesthesia to be effective; therefore the use of these agents to decrease the risk of acid pulmonary aspiration syndrome by no means obviates the need for proper anaesthesia technique during induction of anaesthesia.

Administration, Oral↗

Isolated regional perfusion; anaesthetic technique, monitoring and blood replacement.

Isolated regional perfusion for the treatment of malignant melanoma is an accepted method of treatment. No standard of anaesthetic practice has been established for those individuals. Perioperative records of patients undergoing isolated limb perfusion were studied to determine adequate blood replacement. Records were examined and compared for (1) age, (2) ASA physical status, (3) presence of associated disease, (4) anaesthetic technique, (5) the amount of blood and fluid replacement, (6) preoperative haemoglobin (hgb) and haematocrit (hct) and postoperative serial complete blood counts. Fifteen patients were studied (mean age 53 +/- 16 yrs). Mean blood and fluid replacement was: packed red blood cells; 2.28 +/- 0.82 units, 722 +/- 17 ml of 5 per cent albumin, 1747 +/- 21 ml crystalloid. There were twelve Physical Status Class I or II and three Class III patients. All patients received general anaesthesia. There was a statistically significant difference in the preoperative and postoperative values for haemoglobin and haematocrit (p less than 0.01) with no difference between the postoperative and discharge values. Adequate blood replacement was determined by the equation: (Formula: see text) Extensive invasive monitoring is not routinely required for adequate blood replacement or the detection of leaks between the systemic and isolated circulation.

Adult↗

Early extubation after high-dose fentanyl anaesthesia for aortocoronary bypass surgery: reversal of respiratory depression with low-dose nalbuphine.

To investigate the possibility of selective reversal of narcotic-induced respiratory depression following fentanyl anaesthesia, we studied 20 patients after aortocoronary bypass surgery. All patients were anaesthetized with fentanyl, 40 micrograms . kg-1 and oxygen, with isoflurane as indicated. In a random double blind fashion either incremental doses of nalbuphine, or normal saline were administered approximately four hours after cardiopulmonary bypass. Respiratory depression was evaluated using blood gas and end tidal CO2 (PETCO2) measurement, and in addition, a ventilatory response to CO2 was obtained preoperatively and at selected intervals postoperatively. Despite randomization, patients with more respiratory depression were assigned to nalbuphine. There appeared to be a reversal of respiratory depression with nalbuphine, indicated by a fall in the resting PETCO2 value. This apparent reversal of respiratory depression was associated with a significant increase in pain, requiring treatment in three patients. We conclude that low-dose nalbuphine is not an acceptable method of antagonism of respiratory depression in this group of patients. Many patients who did not receive nalbuphine were able to breathe adequately at an earlier stage than was previously suspected. Close monitoring of the respiratory system may permit earlier extubation without the requirement of a narcotic antagonist after this dose of fentanyl.

Anesthesia, General↗

Sudden infant death syndrome--are parents getting the message?

BACKGROUND: Factors that place an infant at increased risk of sudden infant death include the prone sleep position, overheating and parental smoking, while practices such as bottle-feeding, co-sleeping and the use of pacifiers remain controversial. Major publicity campaigns have been undertaken, which have included the distribution of printed material and extensive media coverage. AIMS: To examine if Irish parents follow the currently recommended childcare guidelines to reduce the risk of sudden infant death and to examine factors that may have impact on their acceptance. METHODS: A random selection of 197 infants from the Birth Register of the Eastern Health Board. Parents were interviewed and a semi-structured survey questionnaire was completed. RESULTS: Forty one per cent of infants are still placed on their side to sleep, an inherently unstable position. First time parents are more likely to place their infants on their backs. Over 60% of infants are exposed to one or more adults smoking in the home despite parental knowledge of its association with sudden infant death syndrome (SIDS). Sixty eight per cent of infants are overwrapped at night and parental understanding of what constitutes overwrapping is poor. Thirteen per cent of infants regularly co-sleep with their parents and 20% of these parents smoke. Pacifier use is common. CONCLUSION: Future programmes should target first time parents, should provide clear information regarding appropriate infant thermal environment, and should ensure regular updating of medical personnel so that they can instruct families on best current practice. Smoking remains a significant health issue with an impact on sudden infant death.

Adult↗