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Lot 89SF has been the reference standard serum pool used in pneumococcal enzyme-linked immunosorbent assays (ELISAs) since 1990. In 2005, it was estimated that there remained between 2 and 5 years' supply of lot 89SF. Since lot 89SF was the reference standard used in the evaluation of the seven-valent pneumococcal conjugate vaccine Prevnar (PCV7), the link to clinical efficacy would be severed if stocks became completely depleted. Furthermore, demonstration of immune responses comparable to those elicited by PCV7 is a licensure approach used for new pneumococcal conjugate vaccines, so a replacement reference standard was required. A total of 278 volunteers were immunized with the 23-valent unconjugated polysaccharide vaccine Pneumovax II, and a unit of blood was obtained twice within 120 days following immunization. Plasma was prepared, pooled, and confirmed to be free from hepatitis B virus (HBV), hepatitis C virus (HCV), and HIV. The pooled serum was poured at 6 ml per vial into 15,333 vials and lyophilized. Immunological bridging of 007sp to 89SF was used to establish equivalent reference values for 13 pneumococcal capsular serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F) by five independent laboratories. Antibody concentrations in 007sp were established relative to the lot 89SF reference preparation using the WHO reference ELISA. Subsequently, 12 existing WHO calibration sera had concentrations reassigned for 13 pneumococcal serotypes using new serum 007sp as the reference, and these were compared to concentrations relative to the original reference serum. Agreement was excellent for the 12 WHO calibration sera. The 007sp preparation has replaced 89SF as the pneumococcal reference standard. Sufficient quantity of this new preparation is available such that, with judicious use, it should be available for at least 25 years.

Diffuse pollution from agriculture remains a significant challenge to many countries seeking to improve and protect their water environments. This paper reviews literature relating to the provision of information and advice as a mechanism to encourage farmers to mitigate diffuse pollution. The paper presents findings from a literature review on influencing farmer behaviour and synthesizes three main areas of literature: psychological and institutional theories of behaviour; shifts in the approach to delivery of advice (from knowledge transfer to knowledge exchange); and the increased interest in heterogeneous farming cultures. These three areas interconnect in helping to understand how best to influence farmer behaviour in order to mitigate diffuse pollution. They are, however, literatures that are rarely cited in the water management arena. The paper highlights the contribution of the 'cultural turn' taken by rural social scientists in helping to understand collective and individual voluntary behaviour. The paper explores how these literatures can contribute to the existing understanding of water management in the agricultural context, particularly: when farmers question the scientific evidence; when there are increased calls for collaborative planning and management; and when there is increased value placed on information as a business commodity. The paper also highlights where there are still gaps in knowledge that need to be filled by future research - possibly in partnership with farmers themselves. Whilst information and advice has long been seen as an important part of diffuse pollution control, increasing climate variability that will require farmers to practice adaptive management is likely to make these mechanisms even more important., (Copyright © 2009 Elsevier B.V. All rights reserved.)

Aims: To evaluate the visual outcome, safety, and economic benefits of immediately sequential phacoemulsification performed under topical anaesthesia as a day case procedure., Methods: Immediately sequential phacoemulsification under topical anaesthesia was performed as a day case procedure on 218 eyes of 109 patients over 21 months by a single senior surgeon using intracameral antibiotics. Case notes were retrospectively analysed., Results: The final best corrected visual acuity (BCVA) was 6/9 or better in 86% of eyes with 47% achieving 6/6 or better. The incidence of intraoperative complications was 0.45%. Postoperative complications occurred in 13.8% of eyes and included refractive error >1.00D from refractive aim (6.9%). Bilateral complications occurred in 5.5% of the patients. There were no cases of endophthalmitis. The mean time from surgery to discharge was 24 days., Conclusion: Immediately sequential phacoemulsification under topical anaesthesia performed as a day case procedure is not associated with an increased number of complications when compared to unilateral phacoemulsification. Under strict surgical protocol with the use of intracameral antibiotics and performed by an experienced surgeon with a good track record this procedure is safe.

Background: The effect of community action on cervical cancer screening rates was explored in 20 rural Australian towns., Methods: Town quarterly Pap test counts, from the Australian Health Insurance Commission, were converted to town rates using Census populations. Pap test rates for 12 quarters before and after mobilization were examined., Results: Wilcoxon-Mann-Whitney tests demonstrated significantly higher per capita Pap test rates pre- to postmobilization for intervention towns compared to matched control towns for women whose last Pap test was more than 2 years ago (P=0.008). Maximum efficiency robust tests examining the equality of mean differences (intervention-control) of per capita rates between pre- and post-mobilization periods also found significant differences for Pap tests where last procedure was between 1 and 2 years ago (R = 13.9, P = 0.039) and near significance for Pap tests where last procedure was less than one year ago (R = 10.6, P = 0.087)., Conclusions: It appeared that underscreened women were more likely and overscreened women were less likely to have a Pap test in response to the intervention. The results indicate that a community action program can positively impact cervical cancer screening rates., (Copyright 2000 American Health Foundation and Academic Press.)

Telemedicine systems aim to provide quality health care services to persons whose access is otherwise restricted by geography and environment. The military medical department has a unique mission to provide all medical care for the battlefields and peacekeeping missions anywhere in the world. In addition, the medical department has to ensure the health of all soldiers, family members, and retirees during peacetime. Hospital closures coupled with a decreased number of military physicians have left many health care beneficiaries without readily available specialty care. They face long waiting lists or incur high out-of-pocket expenses in order to see medical specialists. As a result of the establishment of a virtual Telepain clinic, 56,400 miles were saved in patient and clinician travel. Use of technologies in the emerging field of telemedicine has lead to the creation of numerous military and civilian medical applications such as virtual dermatology, virtual psychiatry, virtual cardiology, virtual nuclear medicine/radiology, virtual pharmacology, and in future, virtual dentistry and ophthalmology.

The most widely used particle dosimetry models are those proposed by the National Council on Radiation Protection, International Commission for Radiological Protection, and the Netherlands National Institute of Public Health and the Environment (the RIVM model). Those models have inherent problems that may be regarded as serious drawbacks: for example, they are not physiologically realistic. They ignore the presence and commensurate effects of naturally occurring structural elements of lungs (eg, cartilaginous rings, carinal ridges), which have been demonstrated to affect the motion of inhaled air. Most importantly, the surface structures have been shown to influence the trajectories of inhaled particles transported by air streams. Thus, the model presented herein by Martonen et al may be perhaps the most appropriate for human lung dosimetry. In its present form, the model's major "strengths" are that it could be used for diverse purposes in medical research and practice, including: to target the delivery of drugs for diseases of the respiratory tract (eg, cystic fibrosis, asthma, bronchogenic carcinoma); to selectively deposit drugs for systemic distribution (eg, insulin); to design clinical studies; to interpret scintigraphy data from human subject exposures; to determine laboratory conditions for animal testing (ie, extrapolation modeling); and to aid in aerosolized drug delivery to children (pediatric medicine). Based on our research, we have found very good agreement between the predictions of our model and the experimental data of Heyder et al, and therefore advocate its use in the clinical arena. In closing, we would note that for the simulations reported herein the data entered into our computer program were the actual conditions of the Heyder et al experiments. However, the deposition model is more versatile and can simulate many aerosol therapy scenarios. For example, the core model has many computer subroutines that can be enlisted to simulate the effects of aerosol polydispersity, aerosol hygroscopicity, patient ventilation, patient lung morphology, patient age, and patient airway disease.

Objective: The aim of the study was to determine whether use of the fetal fibronectin assay would decrease the number of admissions to labor and delivery for diagnosis and treatment of preterm labor., Study Design: A prospective cohort design was used to compare preterm labor admissions during a 12-month period of fetal fibronectin assay use (study) against a baseline period before fetal fibronectin assay was implemented as standard protocol. Patients coming to the physician's office or hospital with signs and symptoms of preterm labor had a sample obtained for fetal fibronectin assay per labeling criteria. Comparisons were made with the Mann-Whitney U test, independent Student t test, chi2 test, and Fisher exact test. P <.05 was considered significant., Results: There was no difference noted in the number of deliveries between the baseline and study years. During the study year 251 of 330 patients evaluated for preterm labor met study criteria and had the fetal fibronectin assay completed. Eight patients did not have fetal fibronectin assay results available because of specimen handling errors, leaving 243 subjects available for study. Compared with the baseline year, the study year had significantly fewer admissions for preterm labor, preterm labor admissions per patient, and prescriptions written for tocolytic agents. In addition, the length of stay per admitted patient was significantly reduced. The study population had no differences in neonatal outcomes from the baseline population in terms of deliveries at <35.0 weeks' gestation, number of admissions to the neonatal intensive care unit, neonatal intensive care unit length of stay, or days of ventilatory support per patient admitted to the neonatal intensive care unit., Conclusions: Use of the fetal fibronectin assay resulted in significantly reduced preterm labor admissions, length of stay, and prescriptions for tocolytic agents. No negative impact on neonatal outcomes was observed. Reductions in admissions for preterm labor and in charges per admission resulted in approximately $486,000 saved during the study period. A trend toward increased corticosteroid administration (for neonates ultimately admitted to the neonatal intensive care unit) was noted.

The 96 well solid phase extraction (SPE) operated by robot in the LC/MS/MS bioanalysis offered rapid sample preparation for drugs and metabolites in biological matrices, based on simultaneous extraction of 96 samples. The use of a disk as sorbent in the 96 well plate further improved the performance of SPE and allowed for small elution volumes, making it possible to 'dilute and shoot" after SPE elution. In this study, a 96 well plate (Empore) was developed, characterized and optimized for several pharmaceutical compounds. In addition, a robot (MultiProbe) was modified to automate the 96 well plate operation. Examples were given to illustrate the major differences of using 96 well disk plate SPE in the method development as compared to the traditional SPE. This technology has been successfully used to support many clinical studies. Typically, a batch of 96 samples were prepared in 1-1.5 hours unattended (except for the replacement of a collection plate). Considerable savings in disposable supplies were also noted.

EBV-induced lymphoproliferative disease (EBV-LPD) is a serious and potentially fatal complication following stem cell transplantation. Strategies have been developed for the cultivation of donor-derived, EBV-specific cytotoxic T lymphocytes (CTL) for stem cell transplant (SCT) patients affected with these disorders, using donor-derived, EBV-transformed B lymphoblastoid cell lines (BLCL) as stimulators. Although cultivation of EBV-transformed BLCL is possible without using an exogenous source of EBV, transformation of autologous B cells with endogenous virus may be slow and inconsistent. Therefore, if exogenous strains of EBV are used to generate BLCL, it may be beneficial to patients to ensure that these cell lines are not producing virus that potentially could be conveyed at the time of CTL infusion. A reliable method of screening for EBV using a cord blood transformation assay has been developed and is described.

SR 49059 ((2S 1-[(2R 3S)-5-chloro-3-(2-chlorophenyl)-1-(3, 4-dimethoxybenzene-sulfonyl)-3-hydroxy-2,3-dihydro-1 H-indole-2-carbonyl]-pyrrolidine-2-carboxamide) is an orally active non-peptide vasopressin V1a antagonist. A sensitive, selective, and robust LC-MS/MS method was developed to determine the plasma and urine concentrations of SR 49059 in support of clinical studies. Plasma samples were prepared based on a rapid extraction procedure using Chem Elut cartridges. The extracted samples were analyzed on a C18 HPLC column interfaced with a Finnigan TSQ 700 mass spectrometer. Positive atmospheric chemical ionization (APCI) was employed as the ionization source. The analyte and its internal standard (2H6-SR 49059) were detected by use of multiple reaction monitoring (MRM) mode. The plasma matrix had a calibration range 0.2-20 ng ml-1, with within and between run accuracy and precision both less than 10%. The chromatographic run time was approximately 3 min. Urine samples were prepared based on a simple dilution with water, followed by analysis under the same conditions as plasma. The calibration range for urine matrix was 20-5000 ng ml-1, with within and between run accuracy and precision less than 11%. The method has been successfully applied to the clinical sample analysis. The plasma assay was also evaluated on a Finnigan TSQ 7000 mass spectrometer. The performance based on precision and accuracy was virtually identical to that on the TSQ 700, with the exception of linearity in calibration curve (the TSQ 700 was linear, the TSQ 7000 was quadratic).

Objective: Our objective was to explore the effectiveness of community action as a strategy for health promotion, through a critical review of five years of community action evaluation literature., Methods: Community action was defined as a health promotion, program that involved the community in implementation and control of the process of the program. Criteria for scientific evaluation of programs were proposed for sampling and control procedures, reliability and validity of instruments, analysis techniques, and specification of details of the intervention. A critical review of the literature, located by an on-line and related reference search, was undertaken for community action aimed at reducing cancer and cardiovascular disease, between January 1990 and May 1995., Results: None of seven community action studies (17 articles) that examined cancer risk factors fulfilled all the criteria for rigorous scientific evaluation. The most methodologically adequate cancer study, the COMMIT intervention, had only a moderate degree of success in reducing community smoking rates. Similarly, none of the six studies (25 articles) on cardiovascular disease fulfilled all the criteria. The results for the most methodologically adequate study, the Minnesota Heart Health Program, were disappointing, with strong secular trends preventing adequate assessment of the intervention effect., Conclusions: The finding that none of the reviewed studies met all evaluation criteria was due to several factors, including political considerations, feasibility, and the continued evolution of the science of evaluation in health promotion. Some important questions are posed for researchers by the failure of methodologically superior projects, such as COMMIT, to show major gains in reducing health risk behaviors.

A comprehensive survey of class I alpha-tubulin (alpha 1) and class II beta-tubulin (beta II) mRNAs was performed using in situ hybridization in order to determine the extent of continued expression of these immature tubulin isotype mRNAs in the adult rat brain. Qualitatively similar distributions of the two isotype mRNAs were observed, with marked variations in hybridization intensity of both probes apparent across different brain regions. Neurons in a wide variety of structures throughout the brain exhibited intense hybridization signals. While the presence of large numbers of neurons with a moderate hybridization intensity could account for the relatively high level of total binding in some regions such as the cerebellar and dentate granule layers, in most cases higher regional mRNA levels reflected greater hybridization intensity per neuron. Little variability in hybridization intensity was typically seen between individual cells within specific nuclei throughout the brain. The presence of occasional intensely labeled neurons scattered throughout the basal ganglia provided the most striking exception to this pattern. While no qualitative differences between the distributions of alpha 1-tubulin and beta II-tubulin mRNAs were observed, consistent differences in the relative intensity of hybridization for alpha 1-tubulin versus beta II-tubulin mRNA were apparent in a few brain regions. Expression by glia did not appear to contribute significantly to detectable levels of either alpha 1-tubulin or beta II-tubulin mRNA. These findings suggest that continued expression of growth-associated tubulin isotype mRNAs may have functional significance in specific neuronal populations of the adult brain. Partial overlap between the distributions of alpha 1- and beta II-tubulin mRNAs and that of GAP-43 mRNA is discussed, as are potential roles for growth-associated tubulin gene expression in supporting cytoskeletal turnover, reactive axonal growth, and dendritic remodeling in the adult brain.

An in situ hybridization study was performed to determine the relationship between levels of mRNAs for the axonal growth-associated alpha 1-tubulin and beta II-tubulin isotypes and the process of collateral axonal sprouting by identified central nervous system (CNS) neurons. A unilateral hypothalamic knife-cut was used to hemisect the hypothalamoneurohypophysial tract, which results in a robust collateral sprouting response by the uninjured neurons of the contralateral supraoptic nucleus (SON) (Watt and Paden: Exp Neurol 111:9-24, 1991). At 10 and 30-35 days after the lesion, cryosections of the SON were obtained and hybridized with 35S-labeled cDNA probes specific to alpha 1- and beta II-tubulin mRNAs. Quantitative evaluation of the resulting autoradiographs revealed that alpha 1-tubulin mRNA levels were significantly increased by 10 days in SON neurons that were undergoing collateral sprouting compared to controls and that this increase was sustained at 30-35 days post-lesion. Less marked increases in hybridization intensity of the beta II-tubulin probe were also apparent in sprouting neurons at both 10 and 30-35 days after the lesion, but were statistically significant only at 10 days. The measured increases in intensity of hybridization of alpha 1- and beta II-tubulin probes are likely to be conservative estimates of the underlying increase in alpha 1- and beta II-tubulin mRNA levels because sprouting SON neurons undergo significant hypertrophy. High levels of both alpha 1- and beta II-tubulin mRNAs were also seen in surviving axotomized SON neurons ipsilateral to the hypothalamic lesion. We conclude that the pattern of regulation of alpha 1- and beta II-tubulin mRNAs in CNS neurons which are capable of supporting new axonal growth includes three elements: maintenance of significant basal alpha 1- and beta II-tubulin mRNA pools in mature neurons, rapid increases in the pool size of the mRNAs following stimulation of collateral sprouting, and sustained elevation of mRNA levels during the period of axonal sprouting.

A sensitive and efficient assay for gamma-aminobutyric acid (GABA) was applied to fresh mouse whole brain synaptosomes where the extracted GABA was analyzed as its di(tert.-butyl(dimethylsilyl)) derivative by gas chromatography-mass spectrometry (GC-MS) using GABA-d6 as an internal standard. Endogenous levels of 20.01 +/- 0.75 nmol GABA/mg protein were found. The method is characterized by a detection limit of about 10 fmol injected GABA derivative and coefficients of intra-day and inter-day variation of 0.95% and 7.7%, respectively. The rate of synaptosomal GABA synthesis was used to determine the activity of glutamate decarboxylase (GAD) as 314.9 +/- 9.0 nmol GABA/mg protein/h. Both GABA levels and GAD activity were significantly elevated by therapeutic doses of the antiepileptic drug valproic acid.

The Toledo Hospital has initiated many innovative quality programs. One initiated with the dietary department involved turning its wholesaler into a partner to provide better-quality products, lower costs and better service. Specific requirements were established in each of these areas including guaranteed mark-ups, price caps and performance reports. The hospital and the wholesaler also do joint negotiating with product manufacturers. Through these and other strategies, the hospital has reduced purchase prices for food items by over 4%, drastically reduced administrative costs of procurement and lead time, increased inventory turns to over 80 and given the dietary department a single resource for "one stop shopping."

By integrating customer satisfaction planning and industrial engineering techniques when examining internal costs and efficiencies, materiel managers are able to better realize what concepts will best meet their customers' needs. Defining your customer(s), applying industrial engineering techniques, completing work sampling studies, itemizing recommendations and benefits to each alternative, performing feasibility and cost-analysis matrixes and utilizing resources through productivity monitoring will get you on the right path toward selecting concepts to use. This article reviews the above procedures as they applied to one hospital's decision-making process to determine whether to incorporate a stockless inventory program. Through an analysis of customer demand, the hospital realized that stockless was the way to go, but not by outsourcing the function--the hospital incorporated an in-house stockless inventory program.

Relationships between the psychological characteristics absorption and neuroticism, and in vitro and in vivo measures of cell-mediated immunity were examined. Thirty-nine female subjects responded to questionnaires, donated blood for analysis of T-cell numbers, and were tested for delayed hypersensitivity skin responses. Consistent with the experimental hypothesis, subjects classified as repressors of negative affect (low absorption/low neuroticism), or extreme expressors of negative affect (high absorption/high neuroticism), showed lower immune responses than other groups of subjects. For the in vitro T-cell measures and the in vivo skin induration measures, there were also pervasive main effects of neuroticism, with subjects higher in neuroticism showing higher immunity than subjects lower in neuroticism.

Objective: The authors hypothesized that psychiatric liaison screening of elderly patients with hip fractures would shorten the average length of hospital stay and increase the proportion of patients who returned home after discharge., Method: The study was performed at Mount Sinai Medical Center in New York and Northwestern Memorial Hospital in Chicago. The subjects were 452 patients 65 years or older who were consecutively admitted for surgical repair of fractured hips. During a baseline year the patients received traditional referral for psychiatric consultation. During the experimental year all the patients at Mount Sinai and the patients on one Northwestern Unit were screened for psychiatric consultation., Results: The patients who received psychiatric liaison screening had a higher consultation rate than those who received traditional consultation. The rates of DSM-III disorders in the experimental year were 56% at Mount Sinai and 60% at Northwestern. The mean length of stay was reduced from 20.7 to 18.5 days at Mount Sinai and from 15.5 to 13.8 days at Northwestern, resulting in reductions in hospital costs ($647/day) of $166,926 and $97,361, respectively. Fees generated from Medicare service delivery could have paid for the $20,000 psychiatric intervention cost at each site. There was no difference, however, between the two years in the discharge placement of patients., Conclusions: Admission psychiatric liaison screening of elderly patients with hip fractures results in early detection of psychiatric morbidity, better psychiatric care, earlier discharge, and substantial cost savings to the hospital.

In a randomised, single-blind, cross-over study, the effects of a high carbohydrate meal or 'no meal' on BP, pulse rate, blood glucose, plasma insulin and catecholamine levels were assessed in eight fit, untreated, elderly hypertensive subjects. A significantly greater fall in supine (P = 0.006) and erect (P = 0.03) systolic blood pressure (SBP) occurred post-meal compared with the no meal phase. The maximum postprandial fall in supine SBP (-24 mmHg, 95% CI -16 to -32 mmHg) occurred at 60 minutes and was associated with a significant rise in supine pulse rate. No overall difference in diastolic BP was seen between the two phases and orthostatic control of BP was maintained. Blood glucose and plasma insulin levels rose after the meal but plasma noradrenaline levels were unchanged during both phases. Thus in these elderly hypertensive subjects the postprandial fall in SBP was not associated with an overall increase in sympathetic nervous system (SNS) activity (as gauged by plasma noradrenaline levels), unlike the findings previously reported in normotensive elderly and young subjects. These changes in BP and the possible impairment of the SNS response to postprandial hypotension should be considered when assessing the BP control, particularly after therapeutic intervention, in elderly hypertensive patients.

The effect of tamoxifen on the vaginal mucosa and on serum oestrone, oestradiol and gonadotrophin concentrations was investigated in a group of nine postmenopausal women with non-metastatic breast cancer. Compared with fit age-matched controls, the vaginal pH in the tamoxifen-treated group was significantly lower and was comparable to levels in fertile women. Two thirds of tamoxifen-treated women had well oestrogenized vaginal smears compared with none in the control group. Follicle-stimulating hormone was significantly reduced, whilst oestrone and oestradiol levels remained unchanged. We conclude, therefore, that tamoxifen has oestrogen-agonistic properties particularly evident in postmenopausal women, even though it is primarily an antioestrogenic drug.

1. The in vivo biliary metabolites of (+/-)-3-dimethylamino-1,1-diphenylbutane hydrochloride (recipavrin) isolated from Wistar rats have been characterized by g.l.c.-mass spectrometry. 2. Non-conjugated metabolites include recipavrin (1), norrecipavrin (2), diphenylbutanone (3), diphenylbutanone oxime (4), diphenylbutanone phenol (12), diphenylbutanone oxime phenol (14), recipavrin phenol (19), diphenylbutanone O-methylcatechol (16) and diphenylbutanone oxime O-methylcatechol (18). 3. Following beta-glucuronidase hydrolysis and extraction from pH 10 solution, diphenylbutanone (3), diphenylbutanone oxime (4), an unidentified compound (6), primary amine (8), norrecipavrin (2), recipavrin (1), phenols (12, 14, 15), norrecipavrin phenol (13), O-methylcatechols (16, 18), diphenylbutanol O-methylcatechol (17), recipavrin O-methylcatechol (19) and a secondary formamide (5) were identified by g.l.c.-mass spectrometry. 4. Various extraction solvents were employed in sample workup. The formamide (5) was present regardless of solvent used, while the trace presence of secondary acetamide (7) may be associated with the use of ethyl acetate. 5. Metabolites isolated after beta-glucuronidase hydrolysis were characterized by g.l.c.-mass spectrometry of the underivatized form, and as the trimethylsilyl (TMS) derivatives, or following methylation with diazomethane or trimethylanilinium hydroxide (TMAH).

The knowledge and estimated retrospective use of postcoital contraception was ascertained from health professionals in Tower Hamlets in the summer of 1988 using a postal questionnaire. Eighty five per cent of general practitioners responded and 91% of these had received requests for postcoital contraception within the previous six months. Only one third of general practitioners had information about postcoital contraception available in their surgeries. Family planning doctors and nurses had the most accurate knowledge of the method but many health professionals appeared to lack sufficient knowledge to ensure appropriate prescribing and to publicize this method to their women patients. It is concluded that if the high rate of abortion in the borough is to be reduced, health professionals as well as women need to be further educated as part of a postcoital contraception publicity campaign. Use of the term 'emergency contraception' rather than the non-medical term 'the morning after pill' may be more effective and reduce the present confusion among both groups.

Valproic acid, an antiepileptic drug, is extensively metabolized in humans. A hexadeuterated valproic acid was used in this study to aid the identification of metabolites and their artifacts in serum and urine extracts. A 600 mg oral dose of hexadeuterated valproic acid was administered to a human volunteer at steady state serum concentrations of unlabelled valproic acid. By using gas chromatography mass spectrometry, metabolites were characterized as their methyl, tert-butyldimethylsilyl, or trimethylsilyl derivatives. A di-unsaturated metabolite with concentrations comparable to 2-ene valproic acid was identified in serum and urine. The presence of a molecular ion doublet in the mass spectrum corresponding to the unlabelled and hexadeuterated dienes reduces the possible structures for this metabolite. A new metabolite, 2-propyl-4-ketopentanoic acid was also identified in serum and in urine. The dicarboxylic acids, 2-propylsuccinic and 2-propylmalonic were identified in the urine extract as metabolites. Identification of isolated metabolites was aided by comparison of the retention times and mass spectra to those of synthesized reference compounds. Diagnostic fragment ions and ion doublets in the mass spectra of the derivatives of newly identified metabolites were compared to those of known metabolites.

Deuterium-labelled methadone and metabolites were used for the g.l.c.-mass spectrometry detection and identification of biliary conjugated methadone metabolites in rats. After beta-glucuronidase hydrolysis the bile extract contained an unknown metabolite that was not ring hydroxylated and retained an intact keto group. Chemical oxidation of the methadone metabolite 2-ethylidene-N,5-dimethyl-3,3-diphenylpyrrolidine, perchlorate salt (EDDP) with m-chloroperbenzoic acid in chloroform, gave a compound identical by g.l.c.-mass spectrometry to the new metabolite. The chemical oxidation product was identified as 2-(4',4'-diphenylheptan-5'-one-2'-yl)oxaziridine by spectroscopic methods. The oxaziridine was shown to quantitatively isomerize to a secondary formamide (2-formamido-4,4-diphenyl-5-heptanone) during g.l.c.-mass spectrometry analysis. The formamide was also isolated by flash column chromatography after reflux of the oxaziridine in m-xylene, and then characterized by spectroscopy. The formamide and oxaziridine g.l.c.-mass spectrometry characteristics were identical. It was concluded on the basis of g.l.c.-mass spectrometry that the metabolite is the secondary formamide.

The latency of the alpha-glucosidase activity of intact rat liver lysosomes was studied by using four substrates (glycogen, maltose, p-nitrophenyl, alpha-glucoside, alpha-fluoroglucoside) at a range of substrate concentrations. The results indicate that the entire lysosome population is impermeable to glycogen and maltose, but a proportion of lysosomes are permeable to alpha-fluoroglucoside and a still higher proportion permeable to p-nitrophenyl alpha-glucoside. Incubation at 37 degrees C in an osmotically protected buffer of of pH 5.0 caused lysosomes to become permeable to previously impermeant substrates and ultimately to release their alpha-glucosidase into the medium. The latencies of lysosomal beta-glucosidase and beta-galactosidase were examined by using p-nitrophenyl beta-glucoside and beta-galactoside as substrates. The results indicate permeability properties to these substrates similar to that to p-nitrophenyl alpha-glucoside. On incubation in an osmotically protected buffer of pH 5, lysosomes progressively released their beta-galactosidase in soluble form, but beta-glucosidase remained attached to sedimentable material. Lysosomal beta-glucosidase was inhibited by 0.1% Triton X-100; alpha-glucosidase and beta-galactosidase were not inhibited.

Average intakes of nonstarch polysaccharides (dietary fiber), foods, and nutrients were measured in representative samples of 30 men aged 50-59 in 4 Scandinavian populations with a 3-4 fold difference in risk for large bowel cancer. The assessment technique, a 4-day weighed record of food consumed and duplicate collections of all food eaten, was validated by chemical analysis of the duplicates, by measuring 24-hour urine and fecal nitrogen excretion, and by comparing the constituents of the urine samples collected during the survey with similar collections 1-2 weeks later. There were good agreements between estimates of fat and protein intake obtained by food-table calculations of the 4-day weighed record and the chemically analyzed duplicates. Urinary plus fecal nitrogen excretion was equal to estimated nitrogen intake during the survey, and no discernable changes in urinary output occurred after the survey, thereby implying that dietary habits had not changed as a result of the investigative technique. It is concluded that the dietary data are indicative of current patterns of food consumption and are sufficiently valid for comparison with data on cancer risk in the 4 areas.

A case of atypical fibromuscular hyperplasia involving the carotid artery in a 38-year-old male is described. The lesion, with associated intimal disruption and subintimal hematoma, caused cerebral embolus and transient neurologic dysfunction. Segmental resection and autogenous graft reconstruction of the carotid artery were successful in removing the lesion and preventing subsequent symptoms.

Epidemiological data from a retrospective analysis of 1307 pathology reports of appendicectomy specimens from four Hunter Valley Hospitals, having a 100% tissue audit, showed that there was a significant difference in normal appendicectomy (histologically normal appendix following appendicectomy for suspected acute appendicitis) rates for the young (male and female 0-12 years) and reproductive female (13-50 years) groups when compared with the elderly (over 60 years) and the rest (male 13-59, female 51-59 years) of the population. The overall normal appendicectomy rate was 26%, and the perforation rate 3.6%. These data suggest that suspected acute appendicitis in the Hunter Valley is managed in an appropriate way. The perforation rate was significantly higher in the elderly group, and management of these patients will remain a problem due to delayed and atypical presentation. Continuing audit programmes and area-wide review should contribute to improved accuracy in the diagnosis of acute appendicitis.

A microculture system for in vitro induction of tumour-specific immunity with syngeneic spleen cells and plasma-cell tumours is described. The system generates cytoxic lymphocytes assessed by their ability to lyse 51Cr-labelled target cells in a microcytotoxicity assay. Both the inductive and effector phases are subject to many variables in material and methodology and the optimal conditions are defined in this study. Of particular importance are the responder- to stimulator-cell ratios, the presence of 2-mercaptoethanol, the day of harvest, the method of target-cell 51Cr labelling, and preincubation of the cytotoxic lymphocytes prior to assay.

A fibre made of psyllium husk was given to 12 elderly patients for 4 months in order to investigate their faecal output and selected serum parameters. The fibre significantly improved bowel function and faecal output confirming its value as a non-irritant, harmless bulk-forming laxative. Serum cholesterol was decreased by 20% while triglycerides remained unchanged. There was a significant reduction in serum calcium after withdrawal of the fibre supplement. There were no significant changes in serum iron, total iron binding capacity, fibrinogen, or in the haematological parameters (ESR, haemoglobin, leucocyte count) attributable to the fibre. The high phytate content of the fibre does not appear to have any clinically significant effect on mineral absorption. It is suggested that dietary fibre (at least psyllium-based fibre preparations) may have significant cholesterol lowering capacity due to the binding of bile-acids in the intestine.

Surgically made double dorsal and double ventral upper arms, comprising double extensor and double flexor muscles respectively, were amputated immediately following operation; both limb types regenerated. In terms of skeletal anatomy, a range of limb types was found. These included four-digit limbs of normal cartilage pattern, and hypomorphic limbs having from one to three digits. All of the double dorsal and the majority of the double ventral limbs were symmetrical in the dorsal-ventral axis. This was detected by analysing their muscle patterns at carpal and mid-forearm level, and muscle and epidermal characteristics in the metacarpal region are discussed in terms of the significance of healing time and stump pattern on the regenerative ability of surgically created limb regions.

Experiments have been performed to assess the potency of idazoxan (RX 781094) at alpha and beta-adrenoceptors and dopamine receptors and on catecholamine uptake processes in rat brain. The effects of idazoxan on the turnover rates of noradrenaline and dopamine have been determined. Radioligand binding studies with cerebral cortex membranes have demonstrated that idazoxan exhibits 46-fold selectivity for alpha 2-adrenoceptors labelled by (3H)-idazoxan (Mean Ki +/- S.E.M. = 3.1 +/- 0.4 nM) compared with alpha 1-adrenoceptors labelled by (3H)-prazosin (Mean Ki +/- S.E.M. = 142 +/- 27 nM). Under the same conditions, yohimbine showed 6-fold selectivity for alpha 2-adrenoceptors. Idazoxan had low affinity for beta-adrenoceptors labelled by (3H)-dihydroalprenolol (IC50 value greater than 10 microM), for dopamine receptors labelled by (3H)-domperidone (IC50 value greater than 20 microM), for the (3H)-noradrenaline uptake site in rat hypothalamus (IC50 = 31 microM) and for the (3H)-dopamine uptake site in rat striatum (IC50 value approximately 800 microM). In rats treated with alpha-methyl-p-tyrosine, idazoxan (10-80 mg/kg, po) produced a marked increase (63% at 10, 217% at 20 mg/kg, po) in the apparent rate of turnover of noradrenaline in rat cortex/striatum, without affecting the rate of turnover of dopamine. This was in contrast to yohimbine (5-20 mg/kg, po) which increased the turnover rates of both catecholamines. In the absence of alpha-methyl-p-tyrosine, idazoxan (5-40 mg/kg, po) produced a dose related increase in the MHPG concentration and a small (20-30%) reduction in the steady state concentration of NA; the duration of the reduction was dose-related. DA steady state concentrations were unaffected. Idazoxan is a new selective alpha 2-adrenoceptor antagonist which should prove a valuable investigative tool in neurochemical studies and which may be a useful clinical agent in the management of the affective disorders.