bluuebunny/crossref_metadata_2025 huggingface dataset:
Feedback of Laboratory Usage and Cost Data to Clinicians: Does it Alter Requesting Behaviour?
Authors: Gama R , Nightingale P G , Broughton P M G , Peters M , Bradby G V H , Berg J , Ratcliffe J G
Published: February 2014

In a 1 year prospective study we evaluated the effect of feedback of laboratory data on the requesting behaviour of physicians in general medicine. Data on within-hours and out-of-hours clinical chemistry laboratory usage and revenue expenditure for inpatients and outpatients, expressed in terms of clinical workload, were supplied monthly to a group of three consultant physicians in general medicine. With these data the physician could monitor his performance over a period of time and compare it with that of his peers. Two consultants in general medicine who received no information served as controls.

Over a period of 6 months, there was a 25%, 13% and 18% decrease in tests ( P < 0·01), requests ( P < 0·05) and revenue expenditure ( P < 0·01) per outpatient visit, respectively, in the intervention group of physicians following the introduction of feedback when compared to their baseline period and to the control group. The decrease ( P < 0·01) was in the commonly requested and ‘seemingly cheap’ tests. There was no significant change in laboratory use and expenditure on inpatients.

The feedback of laboratory data was acceptable to the physicians, raised their awareness of laboratory usage and costs and decreased laboratory workload and expenditure.
Different Effects on Oestrogen Binding Sites and Anti-Oestrogenic Action of Danazol and Progesterone
Authors: Tamaya Teruhiko , Wada Keisuke , Mori Hidehiro , Imai Atsushi
Published: February 2014

Rabbit uterus contains type I and II oestrogen binding sites in the cytosol, and nuclear fractions. Oestrogen-stimulated increase in uterine weight was inhibited by concurrent treatment with progesterone or danazol. Oestrogen-induced type I binding sites were decreased by progesterone and to a lesser extent by danazol, while oestrogen-induced type II sites were decreased almost equally in the two groups. Neither progesterone nor danazol alone caused any detectable changes in uterine weight. These findings may suggest that the anti-oestrogenic effect on uterine weight is more correlated with the change in type II binding sites of oestrogen than that in type I sites. Thus, danazol may exert anti-oestrogenic actions through a pathway independent from that of progesterone.
Nasal Secretions from Patients with Polyps and Healthy Individuals, Collected with a New Aspiration System: Evaluation of Total Protein and Immunoglobulin Concentrations
Authors: Biewenga J , Stoop A E , Baker H E , Swart S J , Nauta J J P , van Kamp G J , van der Baan S
Published: February 2014

This study was designed, first, to test a new system for aspiration of human nasal secretions and, secondly, to evaluate protein and immunoglobulin concentrations in these secretions at different levels of secretory activity. The direct aspiration system combines the advantages of minimal irritation of the nasal mucosa with the facility to determine concentrations per gram of secretion. The total protein and immunoglobulin concentrations were inversely related to the amount of secretion obtained. Variations in fluid secretion throughout the day may be responsible for this relationship. The inverse relationship was much more significant in patients with nasal polyps, in which much higher concentrations were found, than in healthy subjects. Ratios of immunoglobulin to total protein were independent of the amount of secretion obtained. Compared to the controls, the ratios of IgM and IgG to protein in the secretions of the patients were significantly increased. The secretory immunoglobulin A to total protein ratios were only slightly higher in the patients' secretions.
A Sandwich Enzyme-Linked Immunosorbent Assay for β2-Glycoprotein I
Authors: Sansom P A , Marlow C T , Lapsley M , Flynn F V
Published: February 2014

A solid-phase sandwich enzyme-linked immunosorbent assay for determining β2-glycoprotein I in urine has been developed. It has a working concentration range of 5–40 μg/L and a detection limit of approximately 1 · 4 μg/L. The within-plate coefficient of variation (CV) falls between 1 · 4% and 2 · 1%, and the between-batch CV ranges from 5 · 2 to 6 · 0%. Recovery of β2-glycoprotein I added to urine varies between 96 and 110%. The assay can also be used for determining β2-glycoprotein I in serum.
Are Patients with Low Serum Thyroid Stimulating Hormone and Normal Total Thyroxine Hyperthyroid? Usefulness of 99mTc Pertechnetate Uptake
Authors: Jackson R S , Jewkes R , Carter G D , Alaghband-Zadeh J
Published: February 2014

The records of 107 patients who had had thyroid 99mTc pertechnetate uptake measured, were reviewed. In patients with normal serum thyroxine (T4) and thyroid stimulating hormone (TSH), 35 of 36 with normal uptake and three with high uptake were clinically euthyroid. In patients with high serum T4 and low TSH, seven had normal and 29 had high uptake and all were clinically hyperthyroid. In patients with normal serum T4 and low TSH, 10 of 11 with high uptake were clinically hyperthyroid whereas only two of 18 with normal uptake were. The sensitivity, specificity and positive predictive value of pertechnetate uptake measurements were 83%, 94% and 91%, respectively. The measurement of pertechnetate uptake is a rapid investigation and may help in the interpretation of patients with undetectable serum TSH found in the presence of normal serum T4.
Enzyme Immunoassay of Serum Erythropoietin in Healthy Children: Reference Values
Authors: Pressac M , Morgant G , Farmer M A , Aymard P
Published: February 2014

A solid phase enzyme immunoassay for erythropoietin which required only a small specimen of blood was evaluated. It permitted measurement up to 150 IU/L. Recovery was between 86 · 2% and 110 · 0% and between-batch precision between 4 · 7% and 5 · 7%. We determined the levels of erythropoietin in infants aged 10 months (geometric mean 11 · 2 IU/L; 95% range 3 · 6–34 · 9) and in children aged 2 years (geometric mean 10 · 5 IU/L; 95% range 4 · 3–25 · 8). Both were significantly higher than the values found in children aged 4 years (geometric mean 7 · 2 IU/L; 95% range 2 · 4–21 · 8). There were no differences between the values in children at 4 years, those aged 4–15 years and adults.
Increased Urinary Transferrin Excretion in Exercising Normoalbuminuric Insulin-Dependent Diabetic Patients
Authors: O'Donnell M J , Martin P , Cavan D , Parkes A , Chapman J , Chapman C , Barnett A H
Published: February 2014

The median rate of urinary transferrin excretion is greatly increased by exercise in subjects with uncomplicated type 1 (insulin-dependent) diabetes. This increase is proportionally far greater than that seen in urinary albumin excretion rate after the same exercise. Non-diabetic control subjects showed no rise in urinary transferrin excretion rate following exercise. N-acetyl-β-D-glucosaminidase excretion rate was higher in diabetic than control groups but did not rise with exercise.

Our results suggest that patients with apparently uncomplicated diabetes have abnormal renal function. In this group an elevated urinary transferrin excretion rate appears to be a more sensitive indicator of altered renal function than is elevated albumin excretion rate. The mechanism underlying exercise-induced urinary loss of transferrin remains to be elucidated.
Detection of Human Pancreatic Pro-Phospholipase A2 Activation Using an Immunoassay for the Free Activation Peptide DSGISPR
Authors: Gudgeon A M , Patel G , Hermon-Taylor J , Hurley P , Bowyer R C , Jehanli A M T
Published: February 2014

There are several forms of the enzyme phosphoLipase A2 (PLA2) in human tissues. In the pancreas the enzyme is produced as a zymogen, pro-phospholipase A2 (pro-PLA2). The active form is generated upon proteolytic cleavage of the N-terminal prophospholipase A2 activation peptide (PLAP), with the sequence Asp-Ser-Gly-Ile-Ser-Pro-Arg (DSGISPR). Antisera specific for free PLAP were produced by immunization with the synthetic peptide, N-terminally conjugated to bovine thyroglobin. Affinity purified antibodies were used to develop a radioimmunoassay with a detection limit of 5 nmol/L. Competitive inhibition studies with amino-terminally truncated sequences showed that, at least, the C-terminal pentapeptide (GISPR) was required for significant inhibition. Anti-PLAP antibodies did not react with native human pancreatic homogenate (a source of pro-PLA2). A large immunoreactive signal was generated upon trypsinization, which coeluted with synthetic PLAP when cromatographed on Sephadex-G25. Likewise, Sephadex-G50 chromatograph fractions of the untrypsinized homogenate reacted with the antibodies only after trypsinization. The immunoreactive signal appeared at a molecular weight of 14 500 which corresponds to the reported molecular weight of pancreatic pro-PLA2. This demonstrates that the assay is specific for the free peptide and reports pro-PLA2 activation. PLAP assay may therefore contribute to the study of the role of the PLA2 activation event in disease states such as pancreatitis.
Properties of Serum Alcohol Dehydrogenase in Nigerians with Primary Hepatoma
Authors: Ogunleye I O , Olusi S O
Published: February 2014

The serum activity of alcohol dehydrogenase was determined in healthy controls and in patients with liver diseases. The mean activity in hepatoma (6·4 ± 1·0U/L) was significantly higher (P<0·05) than the mean values in liver cirrhosis (2·7 ± 0·5U/L); hepatitis (4·3 ± 1·OU/L), obstructive jaundice (2·9 ± 0·5U/L) and healthy controls (0·7 ± 0·1U/L).

Alcohol dehydrogenase purified by CM-cellulose chromatography from the sera of patients with hepatoma had a higher affinity for butanol long chain saturated and unsaturated alcohols than the purified enzyme from healthy controls. Similarly, hepatoma alcohol dehydrogenase oxidized ethanol very poorly ( KM = 154μM) when compared with that from healthy controls ( KM = 40·2μM). Hepatoma alcohol dehydrogenase was inhibited by pyrazole while those of other liver diseases and the healthy controls were not.

These properties of serum alcohol dehydrogenase may prove useful in the early diagnosis of hepatoma since biochemical changes occur before morphological changes in the development of cancer.
Early Diagnosis of Acute Myocardial Infarction by CK-MB Mass Measurements
Authors: Collinson P O , Rosalki S B , Kuwana T , Garratt H M , Ramhamadamy E M , Baird I M , Greenwood T W
Published: February 2014

We have studied the changes in creatine kinase (CK) and creatine kinase MB (CK-MB) activity and concentration for the diagnosis of acute myocardial infarction in 73 patients admitted to the coronary care unit with cardiac symptoms of 12 h duration or less. Serial blood samples were obtained for an 8 h period following admission and CK, CK-MB activity and concentration measured. We compared the performance of single values at optimized diagnostic cut-offs and incremental change (log slope) for all three measurements. CK slope combined with CK-MB concentration measurements allowed accurate diagnosis at 4 h from admision. CK-MB concentration determination 8 h from admission (12–16 h from the onset of chest pain) was the most efficient single measurement. Rapid diagnostic categorization and possible selection of patients for thrombolysis in patients with an uncertain admission diagnosis is possible by these techniques.
Hypophosphataemia in a Hospital Population and the Incidence of Concomitant Hypokalaemia
Authors: Crook M
Published: February 2014

There have been few recent studies describing the incidence and causes of hypophosphataemia in a hospital population and the frequency of concomitant hypokalaemia. In this study a majority (53%) of patients with hypophosphataemia had been on a glucose infusion at the time of their low plasma phosphate. The largest patient group found to show hypophosphataemia (< 0·8 mmol/L) were in the Intensive Therapy Unit (ITU), and this was also the largest patient group manifesting severe hypophosphataemia (< 0·3 mmol/L). Approximately, 11% of patients with hypophosphataemia (< 0·8 mmol/L) had a plasma potassium equal to or less than 3·5 mmol/L, compared with 5·7% of hospital patients without hypophosphataemia. Furthermore, 20% of patients with severe hypophosphataemia (< 0·3 mmol/L) had hypokalaemia as similarly defined.
Incomplete Testicular Feminization Syndrome: Studies of 17β-Oestradiol-Binding Activity and Aromatase Activity in Cultured Genital Fibroblasts Showing Impaired Dihydrotestosterone-Binding
Authors: Imai Atsushi , Ohno Tsukasa , Nakagawa Miki , Sawairi Miho , Tamaya Teruhiko
Published: February 2014

Dihydrotestosterone (DHT) and 17β-oestradiol binding, and aromatase activity were measured in cultured genital skin fibroblasts from two siblings with the incomplete testicular feminization syndrome. Whole-cell and nuclear DHT binding in the cells from both patients were reduced to 80% and 60%, respectively of those in a normal male subject. The nuclear oestradiol binding was not impaired in the fibroblasts from the patients whose plasma oestradiol was normal or a little elevated. Although gonadectomy led to a decrease in plasma testosterone concentration, the concentration of testosterone-binding globulin showed no change suggesting no apparent effect on testosterone-binding globulin synthesis by testicular steroids. There was no significant difference in aromatase activity of the fibroblasts between the patients and the normal male subjects. The relatively increased oestrogen concentration in the syndrome might result from oestrogen production in testes, but is unlikely to be from peripheral tissues such as fibroblasts. This is the first report to examine oestradiol binding and aromatase activity in the cells from the incomplete testicular feminization associated with impaired DHT binding. These findings may give new insight into the pathogenesis of abnormal male sexual differentiation in the patients with testicular feminization syndrome.
Circulating Levels of Tartrate-Resistant Acid Phosphatase in Macrophage-Activated Lung Disease
Authors: Kehely Anne , Moss Donald W
Published: February 2014

Tartrate-resistant acid phosphatase (TrACP) is abundant in alveolar macrophages, suggesting that these cells might contribute to the activity of this isoenzyme in sera of patients with conditions characterized by activation of alveolar macrophages. TrACP was therefore measured in patients with pulmonary sarcoidosis and cryptogenic fibrosing alveolitis and compared with values in controls. Since osteoclasts are known to be the main source of TrACP in serum several indices of bone-turnover were also measured: Serum bone-specific alkaline phosphatase and urine hydroxyproline: Creatinine ratios. Patients with Paget's disease of bone constituted a reference group presenting increased bone turnover.

TrACP was not significantly higher in the lung-disease groups than in controls, although there was a strong positive correlation with angiotensin-converting enzyme in pulmonary sarcoidosis. As expected, TrACP activity was elevated together with the other indices of bone turnover in Paget's disease. It is unlikely that TrACP from alveolar macrophages contributes significantly to serum acid phosphatase activity in lung disease.
Role of Reactive Oxygen Species in the Generation of Fluorescence by Glycation
Authors: Le Guen C A , Jones A F , Barnett A H , Lunec J
Published: February 2014

Fluorescence (excitation 360 nm, emission 454 nm) generation in glycated albumin was investigated. Antioxidants and the metal chelator desferrioxamine (DFX) were used to study the mechanism of fluorescence generation. Delipidation studies, reverse phase chromatography and scanning fluorimetry were performed to examine the nature of this fluorescence. The mechanism of action of aminoguanidine, a compound which has been shown to inhibit the formation of visible fluorescence in proteins in vitro and in vivo was investigated in relation to glycation and by comparison with compounds with structural similarities.

We conclude that hydrogen peroxide, metal ions and hydroxyl radicals are involved in fluorescence generation in glycated albumin, which is largely lipid in nature, and arises through glycation, amino acid oxidation and changes in bound lipid. Our results suggest that the action of aminoguanidine is not specifically related to blocking of ketoamine groups on glycated proteins as previously suggested.
A Method for Quantitative Measurement of Mitochondrial Creatine Kinase in Human Skeletal Muscle
Authors: Smeitink J , Wevers R , Hulshof J , Ruitenbeek W , Lith T v , Sengers R , Trijbels F , Korenke C , Wallimann T
Published: February 2014

Defects in the mitochondrial energy generating system in patients with a mitochondrial myopathy are known to be localized in various enzyme complexes involved in energy production. Such a defect may exist at the level of mitochondrial creatine kinase (Mi-CK). On that account we have developed a method for measurement of the enzyme activity in human skeletal muscle biopsy material (> 10 mg). Interfering creatine kinase isoenzymes are removed by anion exchange and affinity chromatography. The activity of Mi-CK in reference skeletal muscle homogenates amounts to 240±88 mU/mg protein (30±8·0 mU/mg wet weight).
Immunoblotting of Transferrin in the Identification of Cerebrospinal Fluid Otorrhoea and Rhinorrhoea
Authors: Keir G , Zeman A , Brookes G , Porter M , Thompson E J
Published: February 2014

Cerebrospinal rhinorrhoea is potentially serious due to the risk from infection. In patients presenting with a nasal discharge of clear fluid it is important to identify the nature of the fluid. Cerebrospinal fluid is readily identified by the presence of asialo-transferrin (tau protein). A method is presented for the identification of tau protein based upon agarose electrophoresis, followed by transfer onto cellulose nitrate membrane and immunochemical detection of transferrin. The method is reliable, sensitive and simple, and requires only basic electrophoresis apparatus.
Glomerular and Tubular Proteinuria in Type 1 (Insulin-Dependent) Diabetic Patients with and without Retinopathy
Authors: Martin P , Tindall H , Harvey J N , Handley T M , Chapman C , Davies J A
Published: February 2014

We compared the urinary excretion of albumin, transferrin, N-acetyl-β-D-glucosaminidase and α-1-microglobulin in 78 Type 1 (insulin-dependent) diabetic patients: 39 with retinopathy and 39 without. The two groups were matched for age, sex and duration of diabetes. The patients with retinopathy had increased excretion (median and range) of albumin [1·7 (0·3–399·1) versus 1·0 (0·3–116·6) mg/mmol creatinine, P < 0·05], transferrin [114·2 (4·1–37126·2) versus 33·4 (1·0–4176·7) μg/mmol creatinine, P < 0·01] and N-acetyl-β-D-glucosaminidase [23·8 (1·1–119·1) versus 15·0 (0·1–65·1) μmol/h/mmol creatinine, P < 0·05] but not α-1-microglobulin. Transferrin excretion correlated with albumin excretion. The prevalence of increased transferrin excretion (transferrinuria) was greater than that of microalbuminuria in patients both with and without retinopathy ( P < 0·01 in both cases). Urinary transferrin seems likely to be predominantly of glomerular origin and merits prospective longitudinal evaluation as a potential index of the microangiopathic process.
Evaluation of Capillaries for Ionized Calcium Measurements
Authors: Sachs C , Rabouine P , Kindermans C , Dautzenberg M D , Dechaux M
Published: February 2014

Commercially available capillaries containing calcium-titrated heparinate as an anticoagulant designed specifically for ionized calcium measurements in blood were tested with four serum pools with ionized calcium concentrations adjusted to 0·75, 1·25, 1·50 and 2·50 mmol/L. Although this is the best available anticoagulant for this purpose, the use of these capillaries induced a ±1% alteration of the original concentration around the titration level and changes of −2 to + 10% at pathological values. The amount of heparinate released exceeded the recommended limit of 50 IU/mL of specimen with some variability (6% to 20%). Increasing the amount of anticoagulant with the objective of avoiding magnetic mixing did not seem to be a valid approach. Finally, Radiometer, IL and AVL capillaries gave the best available and acceptable results in terms of alterations in ionized calcium.
Reference Values for Urinary HMMA, HVA, Noradrenaline, Adrenaline, and Dopamine Excretion in Children Using Random Urine Samples and HPLC with Electrochemical Detection
Authors: Fitzgibbon M , FitzGerald R J , Tormey W P , O'Meara A , Kenny D
Published: February 2014

Random urine samples were collected from 305 children aged from birth to 14 years and the values of hydroxymethoxymandelic acid, homovanillic acid, noradrenaline, adrenaline, and dopamine were measured by high-performance liquid chromatography with electrochemical detection. The results were reported relative to the urinary creatinine concentration and the values declined progressively with increasing age for each analyte with the exception of adrenaline. The results for each age group were not normally distributed and all values except outliers were retained in determining the upper reference limits.
Comparison of Seven Formulae and Isoelectrofocusing for Determination of Intrathecally Produced IgG in Neurological Diseases
Authors: Öhman Sten , Ernerudh Jan , Forsberg Pia , Henriksson Annemarie , von Schenck Henning , Vrethem Magnus
Published: February 2014

Seven different formulae and agarose isoelectrofocusing (AIF) using immunolabelling for IgG were compared for their ability to discriminate between intrathecally produced IgG and transudated IgG in cerebrospinal fluid. All reference limits were set to a specificity of 97·5% (reference group, n = 211). The probability of a positive test ( p+) was evaluated for 112 patients with multiple sclerosis (MS), 42 with meningitis, 114 with noninflammatory diseases affecting the central nervous system (CNS), 23 with Guillain-Barré syndrome, and 56 with various diseases not affecting the CNS. Agarose isoelectrofocusing had the best diagnostic sensitivity (93%) for MS, combined with a low p+ (0–19%) for other diseases. Among the formulae, the IgG extended index and Reiber's hyperbolic formula were equivalent, giving high (75–79%) diagnostic sensitivity for MS combined with low p+ (4–22%) for other diseases. All other formulae, although sensitive for MS, had a higher rate of false positive results.
Prevalence of Haemoglobin Variants in a Diabetic Population and Their Effect on Glycated Haemoglobin Measurement
Authors: Allen K R , Hamilton A D , Bodansky H J , Poon P
Published: February 2014

The measurement of glycated haemoglobin (glycated Hb) by automated high pressure liquid chromatography enabled the prevalence of haemoglobin variants (Hb variant) to be determined in a large diabetic population within Leeds, UK. There were 16 patients showing a Hb variant amongst a total of 5300 diabetics, the majority of the variants being HbA/S traits. There were also 13 patients showing a slightly raised HbF in the range of 2% to 5%. A multi-centre study was carried out by distributing blood specimens from some of the patients with Hb traits. Significant differences in the percentage of glycated Hb were observed relating to the methodology used. However, glycated Hb is used to monitor glycaemic control and laboratories must be aware of Hb variant interferences on their methods. Also, screening for Hb variants should be considered for the at risk groups of diabetic patients.
Defective Collagen Metabolism in Saudi Patients with Hernia
Authors: Ajabnoor M A , Mokhtar A M , Rafee A A , Taha A M
Published: February 2014

Fibroblastic cell cultures were derived from the hernial sac and some of the surrounding muscles (rectus sheath, internal oblique, and/or cremasteric muscle) of 130 Saudi patients with different types of herniation, and from 21 control subjects. The rates of cell proliferation were studied for 39 days. Results suggested decreased rates of proliferation of cells derived from patients compared to controls. In vitro studies of the rates of incorporation of 14C proline into the muscle biopsies revealed decreased rates of label incorporation in the samples derived from patients compared to controls. However, no differences were detected between rates of collagenase activities of the biopsies obtained from patients compared to those of controls. These findings suggest that collagen synthesis is probably defective in the studied group of Saudi patients with hernia.
Comparison of Post-Mortem Urinary and Vitreous Humour Organic Acids
Authors: Bennett Michael J , Ragni Marie C , Hood Ian , Hale Daniel E
Published: February 2014

We have analysed organic acid profiles in 74 samples of post-mortem vitreous humour from the suddent infant death syndrome and compared the profiles to those obtained from the corresponding urine or bladder wall swab. There was a high degree of correlation indicating that vitreous humour analysis in high-risk infants is an appropriate analytical strategy when urine is not available. In our patient sample two infants had evidence of abnormal methylmalonic acid metabolism, one had glyceric aciduria (and elevated levels of vitreous humour glyceric acid), one had evidence of pre-existing liver damage as judged by the presence of 4-hydroxyphenyllactic acid, one had a non-ketotic dicarboxylic aciduria indicating inhibited fatty acid oxidation and two patients had significant long-chain 3-hydroxydicarboxylic acids and evidence of paracetamol ingestion.
New Automated Nonisotopic Immunoassays for Free Thyroxin: Effect of Albumin and Thyroxin-Binding Globulin Concentrations
Authors: Zweig Mark H , Csako Gyorgy
Published: February 2014

Recently, nonisotopic (often automated) immunoassays for measuring serum free thyroxin (FT4) have become available. Though more costly than radioimmunoassays, they are considerably more convenient. We studied the influence of endogenous albumin and thyroxin-binding globulin concentration on five automated, nonisotopic methods of measuring FT4 [Enzymun on ES300 (one-step), Stratus I and II (essentially two-step), Delfia (two-step), and IMx (two-step)] in a mixed patient population. We observed that they (a) are influenced very little by endogenous serum binding proteins and (b) seem to have sufficient within-run precision to justify performing single measurements on patients' specimens.
Urinary Dopamine Excretion in Chronic Renal Disease
Authors: McClelland P , Yaqoob M , Bone J M , Thompson P , Roberts N B
Published: February 2014

We have measured unconjugated urinary dopamine and investigated its relationship to glomerular filtration rate in two groups of 22 patients with chronic renal failure matched for age and sex, one group with primary glomerular disease, the other with tubulo-interstitial disease. Urine dopamine excretion was similar in both glomerular and tubulo-interstitial disease groups, and correlated significantly with creatinine clearance. Although urinary protein excretion was significantly higher in glomerular disease, urinary sodium excretion, fractional sodium excretion, urine flow rate and free water clearance were similar in both groups and did not correlate with dopamine excretion. These results suggest that in patients with chronic renal disease, urinary dopamine excretion is mainly under the influence of the glomerular filtration rate, irrespective of the underlying pathology, and therefore interpretation of urinary dopamine excretion requires a knowledge of the patients' renal function.
Rapid Enzyme Diagnosis of Patients with Acute Chest Pain Reduces Patient Stay in the Coronary Care Unit
Authors: Collinson P O , Ramhamadamy E M , Stubbs P J , Rosalki S B , Garrat H M , Mosely D , Evans D H , Fink R S , Baird I M , Greenwood T W
Published: February 2014

We have studied the effect of early exclusion of myocardial infarction using rapid biochemical diagnosis on the management of patients admitted to the coronary care unit of a district general hospital. Diagnosis was based on the rate of creatine kinase increase in serial samples obtained over the 8 h following admission. For an initial 3-month familiarization period serial creatinine kinase results were made available at the end of working day to supplement clinical management, supported by our traditional protocol of admission and daily enzyme determinations. Subsequently, for a 4-month period, the admission to 8 h serial values were provided by 1100 h each day and usually within 24 h of admission. There was a net reduction in length of stay on the coronary care unit to a median 2 days ( n = 66) compared with 3 days ( n = 41) for patients without further cardiac symptoms or electrocardiographic changes suggestive of ischaemia or infarction. This change was significant, P = 0·007, Mann-Whitney U test. Reversion to the original protocol of daily enzyme estimations resulted in an increase in the length of stay on the coronary care unit back to a median of 3 days for this patient group. Rapid diagnostic protocols, applied within routine clinical practice, have the potential for real reduction in coronary care unit stay.
Detection of Colorectal Cancer by Faecal α1-Antitrypsin
Authors: Moran A , Radley S , Neoptolemos J , Jones A F , Asquith P
Published: February 2014

Alpha1-antitrypsin was measured in 5-day faecal collections from patients with colorectal cancer or adenomatous polyps and a symptomatic control group. Collections were homogenized and lyophilized prior to measurement of α1-antitrypsin by radial immuno-diffusion. Colorectal cancer dry weight concentrations were significantly higher than the symptomatic control results, with 57% having results greater than 95% normal values of this control group. There was no significant difference between adenomatous polyp dry weight concentrations and symptomatic control results. Wet weight concentrations were calculated from wet/dry ratios. Colorectal cancer wet weight concentrations were significantly higher than symptomatic control results with 48% having results greater than 95% normal values of this control group. Alpha1-antitrypsin was also measured in non-lyophilized samples from an asymptomatic control group ( n = 39). Colorectal cancer wet weight concentrations were significantly higher than asymptomatic control results, with 62% having results greater than 95% normal values of this control group. Faecal α1-antitrypsin measurement deserves further assessment for use in detecting colorectal cancer.
Urinary Retinol Binding Protein: Stability and Pre-Analytical Handling of Specimens for its Measurement
Authors: Guy J M , McMurray J R
Published: February 2014

The stability of retinol binding protein (RBP) in urine has been reassessed and was found to be pH-dependent, diminishing as urinary pH decreased. When urines were incubated at 4°C for 1 week, statistically significant losses of RBP were observed at pH values less than seven, varying in degree in different urines. At 37 °C some urine specimens showed minor losses when incubated for periods up to 4 h. This was more marked at pH 5 than pH 7 with maximum losses of 26% and 12%, respectively. When adjusted to pH 7, urines showed no significant loss of RBP when stored at 4°C for up to 2 weeks; at −20°C no significant change in RBP concentration was found for at least 14 weeks. To minimize losses of RBP it is recommended that urine pH is adjusted to above seven immediately after voiding.
An Assessment of the Effect of Haematocrit on the Hemocue Blood Glucose Analyser
Authors: Wiener K
Published: February 2014

The HemoCue blood glucose analyser was investigated with particular respect to its susceptibility to influence by the haematocrit of the blood specimen. The red cells and plasma of fresh blood samples were separated and then recombined in proportions to give a range of haematocrit values and the resulting specimens assayed for glucose. The HemoCue exhibited no appreciable influence of haematocrit. The instrument was easy to use, stable and had good precision (CV 1·8%). However, results tended to be a little higher than those produced by a conventional laboratory analyser on whole blood and it is understood that the factory calibration of newer HemoCue instruments has been modified to correct for this.
Laboratory Diagnosis of Phaeochromocytoma: Which Analytes Should We Measure?
Authors: Graham P E , Smythe G A , Edwards G A , Lazarus L
Published: February 2014

There is a diversity of advice in the literature as to which biochemical assays are best suited to the investigation of patients with a suspected phaeochromocytoma. The challenge for the clinical laboratory is to select those assays which detect all phaeochromocytomas, whilst having the lowest incidence of false positive diagnoses. We compared the sensitivity and specificity of a wide range of assays currently used for the biochemical diagnosis of phaeochromocytoma using either specific gas chromatographic-mass spectrometric (GC/MS) or high performance liquid chromatography-electrochemical detection (HPLC/ED) techniques.

Noradrenaline (NA), adrenaline (ADR), dopamine (DA), 3,4-dihydroxyphenylglycol (DHPG), hydroxymethoxymandelic acid (HMMA), normetanephrine (NMET) and metanephrine (MET) were measured in 24 h urine specimens from 20 patients with histologically proven phaeochromocytoma and a large group of patients referred for investigation but subsequently found not to have a phaeochromocytoma. Because phaeochromocytomas are a heterogeneous group of hormone secreting tumours, no single analyte could achieve 100% sensitivity; 100% sensitivity was achieved only when the combination of both NA and ADR or NMET and MET was used. DA, DHPG and HMMA all had poor sensitivities. HMMA had a sensitivity of 70% when using the upper 95% confidence level (48μmol/24 h) of the non-tumour patients as the cut-off. By lowering the cut-off to 35μmol/24 h the sensitivity could be increased to 100% but at the expense of the specificity which was decreased from 98 to 92%. On the basis of this study we recommend the specific measurement of either NA and ADR or NMET and MET as the most suitable analytes for the detection of phaeochromocytoma, and further that, due to its poor specificity, HMMA be abandoned as a suitable analyte.
Urinary Growth Hormone Excretion as Measured by a Sensitive Immunochemiluminometric Assay
Authors: Turner G , Brown R C , Weeks I , Butler G E , Creagh F N , Woodhead J S
Published: February 2014

A sensitive immunochemiluminometric assay with a detection limit of 1 · 1μU/L was developed for the measurement of urinary growth hormone (UGH). The assay was shown to be specific and precise. There was a good correlation between serum growth hormone (GH) and UGH concentrations in 20 patients with acromegaly and six volunteers following an intravenous injection of recombinant GH. We concluded therefore that UGH measurements appear to provide a satisfactory index of GH secretion. The use of the assay in the investigation of growth disorders was assessed. We studied 11 pre-pubertal children, six of normal stature, and five of short stature, over a 6-month period. Sequential fortnightly measurements of UGH were carried out and height velocity was determined. The children of short stature grew at a slower rate and excreted less GH than the children of normal stature. However, we observed considerable within-individual variability in GH excretion in both groups (CV 22–98%). We therefore recommend that sequential UGH analyses should be carried out and the results interpreted in conjunction with growth measurements. However, further investigations into the renal handling of GH are needed to establish optimum sampling regimes.
Comparison of a Commercial Enzymic Kit for Urinary Oxalate Analysis with a High Performance Liquid Chromatographic Method
Authors: Starkey Bryan J , Fry Ian D R
Published: February 2014

A new commercial enzymic kit for urinary oxalate determination has been adapted for use on a centrifugal analyser. It has been evaluated and compared with an established high performance liquid chromatographic (HPLC) procedure developed in our laboratory. Mean recovery of oxalate from urine samples augmented with oxalic acid exceeded 97% by both methods. The precision of the HPLC method was superior to that of the enzymic kit but both methods gave between batch precision values better than CV 12% at low (less than 100μmol/L) oxalate concentrations and better than CV 7% at higher concentrations (greater than 270μmol/L)

Urinary oxalate values obtained with the new enzymic procedure correlated more closely with HPLC values ( r = 0·84) than did values previously obtained using the forerunner of the kit ( r = 0·62) which was known to be susceptible to ascorbate interference. No significant interference from ascorbic acid or from high urinary calcium concentrations could be demonstrated using either the improved kit or the HPLC procedure. Its easy adaptation to automated analysers available in most laboratories, coupled to its acceptable analytical performance render the enzymic kit a reasonable alternative to HPLC or other more complex procedures for urinary oxalate analysis.
High-Sensitivity Electrophoretic Method for the Detection of Bence Jones Protein and for the Study of Proteinuria in Unconcentrated Urines
Authors: Aguzzi F , Gasparro C , Bergami M R , Merlini M
Published: February 2014

The study of proteinuria, and especially the search for Bence Jones proteins (BJP), has almost always required urine concentration. We evaluated a high-sensitivity electrophoretic method using unconcentrated urine based on colloidal gold staining. The sensitivity for the detection of BJP is further enhanced by immunofixation. Sensitivity to BJP is better than 1 mg/L and, apart from the α-1 microglobulin, all the proteins relevant to the classification of proteinuria can be visualized with a sensitivity of approximatively 3 mg/L.
Biological Variation of Acute Phase Proteins
Authors: Clark Geraldine H , Fraser Callum G
Published: February 2014

The analytical, within-subject and between-subject components of variation were estimated for serum albumin, transthyretin, α1-acid glycoprotein, α1-antichymotrypsin, haptoglobin, β2-microglobulin and C-reactive protein in a cohort of 19 apparently healthy subjects over 20 weeks. Desirable analytical goals based on biological variation should be able to be met except for serum albumin and β2-microglobulin for which methodological improvement is warranted. All proteins showed marked individuality which casts doubt on the utility of conventional population-based reference values as interpretative criteria. The critical differences required for significance of changes in serial results differ markedly from protein to protein and the data presented allow generation of objective criteria for monitoring individuals.
Improved Formulae for the Judgement of Intrathecal Produced IgA and IgM in the Presence of Blood CSF Barrier Damage
Authors: Öhman Sten , Ernerudh Jan , Forsberg Pia , von Schenck Henning , Vrethem Magnus
Published: February 2014

In order to discriminate between transsudated and intrathecally produced IgA and IgM in patients with blood cerebrospinal fluid (CSF) barrier (BCB) damage, we developed extended indices (IgA-EI and IgM-EI) for these immunoglobulins according to the general formula for extended index of a component X in CSF:

where a is a parameter specific for X. For IgA parameter a was found to be 1·15 and for IgM 1·9.

A preliminary evaluation of IgA-EI and IgM-EI indicated lower false positive rates as compared to CSF IgA and IgM concentrations as well as ‘conventional’ IgA and IgM indices in cases with BCB damage, and essentially the same rates as for the hyperbolic formulae of Reiber and Felgenhauer.

The importance of reliable sampling and analytical technique for IgM in CSF is discussed.
Proteolytic Activity and Free Amino Acid Concentrations in Polymorphonuclear Leucocytes
Authors: Carrea A , Canepa A , Perfumo F , Ancarani P , Verrina E , Gusmano R
Published: February 2014

Polymorphonuclear granulocytes (PMN) are valuable tools for evaluating amino acid (AA) metabolism in nucleated cells, although variations of free amino acid concentrations due to the methods used for the separation of the cells and the procedures used for lysis have been reported. Furthermore, analytical variations in PMN AA concentration may be induced by protease activation during preparation, so that free AA detected in cells could originate from proteolysis other than from the physiological metabolic pathways and transport systems. To study this possibility we measured granulocyte protease activity and AA concentrations in cell suspensions processed with and without the addition of antiproteolytic agents. Granulocyte AA concentrations and protease activity in samples treated with antiproteolytics were 8–15 times lower than in samples processed without antiproteolytics. The use of protease inhibitors throughout the sample preparation is necessary for reliable estimation of free AA in granulocytes.
Assessment of Atypicality: An Adjunct to Prenatal Screening for Down's Syndrome That Facilitates Detection of other Abnormalities
Authors: Wright D E , Reynolds T M , Donovan C M
Published: February 2014

Current software used for assessment of the risk of Down's syndrome may give misleading risk estimates if applied to other abnormalities. Often the abnormality is reflected in maternal serum α-fetoprotein and human chorionic gonadotrophin levels and is then translated into a low risk for Down's syndrome that may not be recognized as significantly atypical of normality. We regard this as a serious deficiency in the current Down's syndrome risk reporting algorithm, and suggest a modification that allows the problem to be overcome.
Differences between the Sugar Moieties of Liver- and Bone-Type Alkaline Phosphatases: A Re-Evaluation
Authors: Miura Masakazu , Sakagishi Yoshikatsu , Hata Keishi , Komoda Tsugikazu
Published: February 2014

We re-evaluated the differences between the sugar moieties of liver and bone alkaline phosphatases (ALPs). Sialic acid was added to ALP sugar moieties by α2,3- or 2,6-sialyltransferase treatment of the asialo-form ALP (neuraminidase-treated ALP). Asialo-bone ALP was converted to a liver-like ALP by the 2,6-sialyltransferase treatment. The resulting liver-like ALP was less susceptible to neuraminidase than non-treated bone ALP, but was still labile to heat exposure at 56°C like non-treated bone ALP. However, after the O-linked sugar moiety had been released by additional treatment with O-glycanase the liver-like ALP became more heat stable at 56°C, like non-treated liver ALP.

Non-treated liver ALP reacted specifically with anti-liver ALP monoclonal antibody, and non-treated bone ALP reacted with both anti-liver and anti-bone ALP antibodies. The asialo-bone ALP still reacted with anti-bone ALP antibody, whereas the asialo-form liver ALP showed little, if any, reaction with anti-liver and anti-bone ALP antibodies. Neuraminidase and O-glycanase-treated bone ALP reacted less with anti-bone ALP antibody. After O-glycanase treatment, bone ALP molecules deprived of an O-linked sugar moiety had a molecular size and heat stability similar to liver ALP. The difference between liver and bone ALP molecules may be due not only to their manner of sialic acid linkage but also to the attachment of the O-linked sugar moiety.
Neonatal Screening for Phenylketonuria: Evaluation of an Automated Enzymatic Method
Authors: Keffler S , Denmeade R , Green A
Published: February 2014

An enzymatic assay for phenylalanine using phenylalanine dehydrogenase is available in kit form (Quantase™) for use with dried blood spot specimens and microtitre plates. The method has been automated by use of a sample processor and its performance and suitability for neonatal screening for phenylketonuria has been evaluated by comparison with the Guthrie bacteriological inhibition assay. The enzymatic method performed well with regard to precision and accuracy. It was able to differentiate between normal and raised phenylalanine concentrations for the purpose of screening, thus greatly reducing the repeat rate for equivocal results. There were, however, some concerns about its robustness for screening and its detection limit. The Quantase assay has the potential to be used as a large-scale routine neonatal screening method, if its use can be shown to be cost-effective.
Development of an Enzyme-Mediated Assay for Phenylalanine in Blood Spots
Authors: Campbell R S , Hollifield R D , Varsani H , Milligan T P , Brearley G , Price C P
Published: February 2014

We describe the optimization of a rapid procedure for the elution of phenylalanine from blood spots and the estimation of the amino acid eluted using an enzyme-mediated assay linked to a colorimetric detection system. The method is rapid, fully quantitative, interference-free, accurate and precise unlike many of the methods currently employed for phenylalanine determination. The performance of the proposed method may necessitate the review of reference ranges and cut-off assignment strategies.
The Use of Cytokine and C-Reactive Protein Measurements in Cerebrospinal Fluid during Acute Infective Meningitis
Authors: Stearman M , Southgate H J
Published: February 2014

Cerebrospinal fluid (CSF) concentrations of C-reactive protein (CRP), tumour necrosis factor α (TNF), interleukin-6 (IL-6), total protein (TP) and white cell count with differential (WCC) have been measured in 24 patients presenting with acute bacterial or viral meningitis and also in a non-infected, non-inflammed control group ( n = 24). In acute viral meningitis, CRP levels were not raised when compared to controls and there was a discordance between high levels of the primary inflammatory mediators (IL-6 and TNF) and the low measured CRP levels. CRP levels were raised in cases of bacterial meningitis. A concentration of 100 ng/mL CRP had a sensitivity of 87% for bacterial meningitis. TNF concentrations in the CSF were significantly raised in cases of acute bacterial meningitis ( P < 0·001). Smaller but variable elevations were seen in the patients with acute viral meningitis. One patient, who succumbed to bacterial infection, showed low CSF levels of CRP, TNF and WCC but an elevated IL-6 concentration. Another, presenting with low CSF WCC, had raised concentrations of CRP, TNF and IL-6 which pointed to the correct diagnosis of acute bacterial meningitis. The development of methods yielding rapid analysis for these cytokines together with a sensitive assay for CRP in CSF would be a useful adjunct to conventional investigation.
Mucopolysaccharidoses Screening: Dimethylmethylene Blue versus Alcian Blue
Authors: de Jong J G N , Heijs W M J , Wevers R A
Published: February 2014

The dimethylmethylene blue (DMB)-based screening procedure for mucopolysaccharidosis1,2 and the alcian blue3 (AB)-based procedure both measure glycosaminoglycan (GAG) content directly in urine. We compared the two procedures. Absorbance per microgram of GAG for DMB was 25 times that obtained with AB, resulting in a requirement for 10 times smaller sample volumes for the former. Recoveries of added heparan sulphate for the AB assay in the absence and presence of added protein (2·5 g/L) were comparable (78–95% and 75–111%), as was the case with the DMB assay. Here, recoveries were generally better (94–103% and 96–100%). The coefficient of correlation (CC) between the two methods measured on 86 reference urine samples was 0·861. For mucopolysaccharidosis urines the CC was higher (0·928). Age-dependent reference values were comparable. Performance of the assays was compared by measurement of GAG content in 24 mucopolysaccharidosis urines. Standard deviation scores (measured value minus mean of reference values divided by standard deviation in reference values) and probabilities for classification in the patient group were calculated and compared. Especially for urines from patients with less distinctly increased GAG content (Sanfilippo, Scheie), scores and probabilities were higher with the DMB assay.