bluuebunny/crossref_metadata_2025 huggingface dataset:
Euthyroid Sick Syndrome in Pulmonary Tuberculosis before and after Treatment
Authors: Chow C C , Mak T W L , Chan C H S , Cockram C S
Published: February 2014

Alterations of circulating thyroid hormones are frequently present in chronic nonthyroidal illnesses and may predict prognosis. Pulmonary tuberculosis, a common treatable debilitating disease, may provide a useful model for detailed evaluation of changes of thyroid hormones in relation to subsequent recovery or mortality. Over a period of 12 months, we performed a prospective study of 40 consecutive Chinese patients aged over 50 years and admitted with newly diagnosed pulmonary tuberculosis. Blood samples were drawn for serial thyroid function tests [free thyroxine (T4), free triiodothyronine (T3) and thyroid-stimulating hormone] before treatment and at 1, 2 and 4 months afterwards. Mortality was determined up to 12 months of follow-up. The euthyroid sick syndrome occurred in 63% of patients at presentation. Twelve of 25 euthyroid sick patients died as compared to one of 15 patients with normal baseline thyroid function tests ( P < 0·02). Among euthyroid sick patients, those who died had significantly lower free T3 concentration at presentation than those who survived ( P < 0·05). An undetectable free T3 concentration at presentation was associated with a subsequent mortality of 75% (9 of 12). Of the survivors, all patients demonstrated a significant rise in serum free T4 concentrations following treatment, which was apparent by 1 month. These data suggest that an undetectable free T3 concentration at presentation reflects severity of illness and predicts a subsequent high mortality.
Proton Nuclear Magnetic Resonance Lipid Profiling of Intact Platelet Membranes
Authors: Yeboah F A , Adosraku R K , Nicolaou A , Gibbons W A
Published: February 2014

A combination of high-resolution one-dimensional and two-dimensional proton nuclear magnetic resonance spectroscopic methods and high pressure liquid chromatography have been used to analyse qualitatively and quantitatively the phospholipids, cholesterol and fatty acid composition of normal human blood platelets. The major phospholipids present were identified and quantified. Information on the nature and composition of the different classes of structural phospholipids and unsaturated fatty acids was obtained and was similar to that reported by others using different analytical procedures. This work showed that the proton nuclear magnetic resonance spectroscopy approach to lipid analysis could provide rapid, comprehensive and detailed information on the roles of lipids in pathological conditions in which platelets have been implicated.
Induction of M-CSF Receptor and its mRNA, and Activation of Tyrosine Kinase in Peripheral Monocytes by Oestradiol-17β and Progesterone
Authors: Fujimoto Jiro , Hori Masashi , Ichigo Satoshi , Itoh Toshiya , Saio Masanao , Takami Tsuyoshi , Tamaya Teruhiko
Published: February 2014

In monocytes at the secretory (oestrogen-progesterone dominant) phase of the menstrual cycle, expression of c-fms and macrophage colony-stimulating factor (M-CSF) receptor and activity of tyrosine kinase (TK) were increased by oestradiol with or without progesterone. In vivo, oestrogen may induce expression of c-fms and M-CSF receptor as well as the activation of TK in monocytes under the milieu of the secretory phase. Alternatively, cells of monocyte lineage during the secretory phase might, via various factors, obtain the potency to induce the expression and the function of M-CSF receptors, this potency being effected by oestrogen. Macrophages in peritoneal fluid in pelvic endometriosis (oestrogen predominant) might be activated during the secretory phase of the menstrual cycle, causing infertility.
Mass versus Activity: Validation of an Immunometric Assay for Bone Alkaline Phosphatase in Serum
Authors: Price C P , Mitchell C A , Moriarty J , Gray M , Noonan K
Published: February 2014

A detailed investigation of the performance of an immunometric assay for the quantitation of bone alkaline phosphatase (ALP) in serum showed that precision of this assay was similar to that of lectin precipitation and electrophoretic methods. Relationships for mass and activity measurements of the bone isoform using samples from children and patients with Paget's disease were similar for lectin and electrophoretic methods. Purified liver enzyme showed 100% cross reactivity in the immunometric assay. However, comparison of the slopes between bone ALP mass and total ALP activity using only samples with predominantly liver or bone isoforms showed that the cross reactivity of the liver isoform in serum was 18·3%. Experiments in which increasing amounts of a sample containing 90% of liver isoform adding to a serum sample from a patient with Paget's disease showed a cross reactivity of 16·5%. The reference range for bone ALP mass was 7–28 μg/L for men ( n = 77) and 5–20 μg/L for women ( n = 110) in the age group 20–50 years. In women over 50 years bone ALP was 28% higher. Increased bone ALP mass was also demonstrated in patients with Paget's disease ( n = 59), liver disease ( n = 95), chronic renal failure ( n = 41) and hyperthyroidism ( n = 17).
The Effects of Gestation Dating on the Calculation of Patient Specific Risks in Down's Syndrome Screening
Authors: Bishop Janine , Dunstan Frank D J , Nix Barry J , Reynolds Timothy M
Published: February 2014

In Down's syndrome screening using biochemical markers, the marker concentrations are adjusted for the gestational age of the fetus, since they are known to change with gestational age. This adjustment is performed by referring to the population median of each marker for the appropriate gestational age group. The measurement of gestational age is subject to error, whatever method is used, and the population median used is actually the median of a mixture of distributions for different true gestational ages. We show how the proportions in this mixture can be estimated and how the true median corresponding to a given true gestational age can be estimated. For simplicity, we consider the case of using a single marker, namely maternal serum α-fetoprotein, and show that the usual estimation method has considerable bias. The effect of this mixture on the calculation of patient-specific risks is discussed and we show that detection rates can be improved by allowing for this error in the dating process. The overall detection rate is increased by about 1%. The increase in detection rate is age-dependent and for some maternal ages the increase is of the order of 5%. The comparative effects of different methods for dating are discussed.
Oestrogen Induces C-Ha-Ras Expression in the Fibroblasts Derived from Human Uterine Endometrium
Authors: Fujimoto Jiro , Ichigo Satoshi , Hori Masashi , Morishita Shigeo , Tamaya Teruhiko
Published: February 2014

In this preliminary study, fibroblasts derived from uterine endometrium as a substitute for normal endometrial stroma were used to analyse the stromal role in uterine endometrium, which depends on oestrogen for growth. C-Ha- ras expression and tyrosine kinase (TK) activity in the fibroblasts were increased by oestradiol, and the increase was diminished by progesterone. Epidermal growth factor (EGF), an activator of TK, also increased c-Ha- ras expression. The combination of both oestradiol and EGF at maximum effective concentration exerted no additive and synergistic effect on induction of c-Ha- ras expression. Pretreatment with tyrphostin, an inhibitor of TK, abolished the oestrogen-inducible expression of c-Ha- ras. Oestrogen might lead to rapid induction of c-Ha- ras expression in endometrial stroma, at least in part, via the activation of TK for the early stages of oestrogen dependent growth in endometrium.
Influence of Race, Age and Sex on the Lymphocyte Subsets in Peripheral Blood of Healthy Malaysian Adults
Authors: Choong M L , Ton S H , Cheong S K
Published: February 2014

The lymphocyte subsets in the peripheral blood of healthy Malaysian adults (212 subjects, age 18–71 years) were analysed using a flow cytometer FACScan in an effort to establish a reference range for the lymphocyte subsets. The lymphocyte subsets studied were T cells (CD3), B cells (CD19), natural killer (NK) cells (CD3−CD16+/CD56+), helper/inducer cells (CD4), cytotoxic/suppressor cells (CD8) and the helper/suppressor ratio (CD4/CD8). The distributions of T cells, CD4 cells and CD8 cells were symmetric about their means while B cells, NK cells and CD4/CD8 ratio followed a skewed distribution. Differences in race were observed for T cells, NK cells, CD4 cells and CD4/CD8 ratio where the Indians were significantly different from the Malays and the Chinese (higher T cells, CD4 cells and CD4/CD8 ratio and lower NK cells). The B cells were significantly lower in the Chinese than the Malays and the Indians. Age differences were seen only in the Chinese where increased CD4 cells and CD4/CD8 ratio, and decreased CD8 cells were observed. A sex difference was observed only in the Chinese where the CD4/CD8 ratio was significantly higher in females than males.
Relationship between a Mild α1 Proteinase Inhibitor Deficiency and Respiratory Symptoms in a Family
Authors: Huet-Duvillier Guillemette , Balduyck Malika , Watrigant Yves , Sesboue Richard , Thiebaut Carole , Lafitte Jean Jacques , Degand Pierre
Published: February 2014

A 34-year-old man with pulmonary emphysema was found to have a mild α1 proteinase inhibitor (α1 PI) deficiency. α1 PI status was investigated in this patient and in 35 members of his family. The α1 PI investigations included α1 PI concentration and phenotype and serum inhibitory capacity for trypsin and pancreatic elastase. Fifteen members of the family had α1 PI concentration and inhibitory capacities below the lower normal limit. Five of these members were characterized by the heterozygous MP phenotype and the 10 others by an apparently homozygous M phenotype, in which the M allele may be associated with another unidentified deficiency allele. Two members of the family had α1 PI concentration and elastase inhibitory capacity below the lower normal limits and trypsin inhibitory capacity within the normal range. They were both characterized by the MP phenotype. Six of these 17 members (three of PI type M and three of PI type MP) showed chronic pulmonary symptoms, whereas among the 19 α1 PI non deficient members, no member had a history of significant pulmonary symptoms.
Urinary Heparan Sulphate Proteoglycan Excretion is Abnormal in Insulin Dependent Diabetes
Authors: Shield J P H , Carradus M , Stone J E , Hunt L P , Baum J D , Pennock C A
Published: February 2014

Urinary excretion of heparan sulphate proteoglycan (HSPG), the main anionic component of the glomerular basement membrane (GBM), was estimated in 30 adolescents and young adults with insulin dependent diabetes (IDDM), 10 with microalbuminuria and 20 sex matched, diabetic controls of similar age without evidence of microalbuminuria. A further 10 non-diabetic control subjects were also examined. Both groups of patients with diabetes had significantly elevated excretion of HSPG when compared to normal individuals. There was no difference in HSPG excretion between diabetic subjects with and without microalbuminuria.
A New Enzymatic Method for the Determination of Inulin
Authors: Sugita O , Tomiyama Y , Matsuto T , Okada M , Gejyo F , Arakawa M , Takahashi S , Watazu Y , Kaneda N
Published: February 2014

A new enzymatic method for the determination of inulin in plasma and urine, using inulase (EC 3.2.1.7), fructokinase (EC 2.7.1.4), phosphoglucoisomerase (EC 5.3.1.9) and glucose-6-phosphate dehydrogenase (EC 1.1.1.49) is described. The assay is based on the hydrolysis of inulin or Inutest (Inutest® which is the injectable form of inulin), by inulase and the determination of fructose released.

The assay was linear up to 2 g/L of Inutest. The within-batch and between batch coefficients of variation were 2·3% and 2·2%, respectively. Recovery of added Inutest from plasma and urine was 98–102%. There was no interference from glucose (27·7 mmol/L), fructose (1·7 mmol/L) or mannose (1·7 mmol/L).

When inulin clearance (using this method) and thiosulphate clearance were compared in 37 patients the inulin clearance was 9·3 mL/min (12%) lower than the thiosulphate clearance. We conclude that this enzymatic method is a simple and specific method.
Serum Binding of Steroid Tracers and its Possible Effects on Direct Steroid Immunoassay
Authors: Micallef Jacob V , Hayes Margaret M , Latif Abdul , Ahsan Rukhsana , Sufi Saulat B
Published: February 2014

We studied the serum protein binding of 3H-labelled progesterone, oestradiol and testosterone, and five 125I-labelled analogues of these steroids. All tracers investigated appeared to be bound by proteins in every serum sample tested. The addition of blocking agents caused a substantial reduction in serum protein binding of 3H-labelled steroids, but had relatively little effect on the binding of analogue steroid tracers. Use of analogue steroid tracers in conventional direct immunoassays for oestradiol and progesterone produced anomalous results for some patient samples when compared to extraction radioimmunoassays, but assays where tracer binding to serum constituents was prevented by adoption of two-step procedures appeared to avoid anomalous results. The results suggest that serum protein binding of steroid analogue tracers may be a source of interference in some direct steroid immunoassays.
Evaluation of an Enzyme-Immunometric Assay for Serum α-Glutathione S-Transferase
Authors: Rees Gareth W , Trull Andrew K , Doyle Sean
Published: February 2014

A commercially available enzyme-immunometric assay for serum α-glutathione S-transferase (GST) was evaluated. Endogenous serum α-GST diluted linearly within the calibration range. However, we recommend that the sample and second antibody reagent are always added sequentially in the assay to avoid hook effect. Between-assay variability was below 7% across the calibration range and the upper limit of the reference range in adults ( n = 219) was 11 · 4 μg/L. Within-individual variability in serum α-GST concentrations measured over a 4–6 week period in 4 healthy adults was small. Serum α-GST concentrations did not change significantly 6 h after a therapeutic dose of paracetamol.

Studies in two patients after liver transplantation showed that serum α-GST is a better discriminant of acute changes in liver function than conventional tests. Serum α-GST concentrations were unaffected by gross muscle damage, extra-hepatic inflammation, or haemolysis and thus appear to be more liver specific than transaminase activities. The effect of renal impairment on serum α-GST concentrations requires further investigation.
Biological Variation in Urinary Excretion of Pyridinium Crosslinks: Recommendations for the Optimum Specimen
Authors: Panteghini M , Pagani F
Published: February 2014

We assessed the analytical and biological variation of pyridinium crosslinks in early morning, 2 h fasting, and 24 h urine specimens from 14 healthy adults over a 1 month period. The results were expressed both in terms of pyridinoline concentration and pyridinoline/creatinine ratio. The data obtained were used to select the optimum specimen for clinical purposes. We found that: ( a) early morning specimens are preferred; ( b) results should be expressed as pyridinoline/creatinine ratio; ( c) reference intervals should be stratified according to gender; ( d) the necessary analytical imprecision (CV≤ 9%), derived from biological variation, is not easily achieved by current methods; ( e) the difference between serial results from an individual must be > 50% to be statistically significant; and ( f) assessment of risk for osteoporotic fracture by means of the pyridinium crosslink assay would, in a significant number of patients, require analysis of multiple urine specimens.
Acidic Catecholamine Metabolites and 5-Hydroxyindoleacetic Acid in Urine: The Influence of Diet
Authors: Mashige Fumiko , Matsushima Yoshikazu , Kanazawa Hideko , Sakuma Ichiro , Takai Nobuharù , Bessho Fumio , Ohkubo Akiyuki
Published: February 2014

Concentrations of vanillylmandelic acid (VMA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), vanillic acid (VA) and 5-hydroxyindoleacetic acid (5-HIAA) in urine from healthy subjects were determined by a high-performance liquid chromatography system with a mixed-mode (C18/ anion-exchange) column and an 8-channel electrochemical detector, in order to study the influence of diet, diurnal variation and age. The urinary excretion of 5-HIAA increased significantly after eating banana, pineapple, tomato, kiwi fruit and walnut. An increase in the urinary excretion of DOPAC and HVA after eating banana and that of VA after taking vanilla was also noted. The urinary excretion of VMA was not significantly influenced by any of the foods examined. The urinary excretion of 5-HIAA in the first-morning urine increased beyond the upper limit of the reference value when banana was taken at 2000 h the previous day. The excretion of all metabolites in the second-morning urine in the fasting state was within respective reference ranges. Diurnal variation of the excretion of VMA, DOPAC, HVA and 5-HIAA in urine was relatively small, but that of VA was large. The concentrations (mmol/mol creatinine) of VMA, DOPAC, HVA, 5-HIAA and VA in the first-morning urine from healthy subjects increased from 7 days after birth to 1 year and then decreased to adult levels at 13 years of age.
Adjustment of Serum Total Calcium for Albumin Concentration: Values Change with Age in Women but Not in Men
Authors: Payne R B , Barth J H
Published: February 2014

A total of 669 women and 609 men were selected from a laboratory computer data base in such a way that they would be expected to have a low prevalence of disturbances of calcium homeostasis but a wide range of serum albumin concentrations. The least squares regression coefficients of total calcium on albumin did not differ between men and women, nor did they differ at different ages. Mean serum albumin-adjusted calcium concentrations did not change with age in men from 1 to 90 years, and values were similar in women aged 1–20 years. However, adjusted calcium concentrations were significantly lower in women aged 21–50 and higher in women aged 61–90 than in younger women and all men. The differences were small and are unlikely to affect clinical interpretation.
The Effects of Cigarette Smoking and Alcohol Consumption on Blood Lipids: A Dose-Related Study on Men
Authors: Whitehead T P , Robinson D , Allaway S L
Published: February 2014

The separate and joint effects of cigarette smoking and alcohol consumption on serum concentrations of total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and triglycerides were investigated in 46 750 men attending the BUPA Health Screening Centre in London during the period 1983–1987, after allowing for differences in age, body mass index and exercise level. Drinking alcohol was found to raise both total cholesterol and HDL-C concentrations, in such a way that HDL-C as a percentage of total cholesterol increased with increasing alcohol consumption. LDL-C concentrations increased with increasing alcohol consumption in non-smokers, but decreased in those smoking over 10 cigarettes per day. Drinking had no significant effect on triglycerides except at high levels of consumption. Smoking raised total cholesterol, LDL-C and triglycerides, but lowered HDL-C concentrations. In particular, smoking even small amounts could negate any protective benefit in HDL-C concentrations gained from moderate consumption of alcohol. Assuming a desirable lipid profile to consist of low total cholesterol, LDL-C and triglycerides and a high HDL-C value, this is best achieved for men by being a non-smoking moderate drinker.
Differences in Serum Luteinizing Hormone Measurements by Immunoradiometric Assay Induced by Kinetic Manipulation of Assay Conditions are Dependent on the Endocrine Milieu of Serum
Authors: Olukoga A O , Mitchell R , Walton L , Robertson W R , Laing I
Published: February 2014

Divergent estimates for luteinizing hormone (LH) in individual serum samples may be given by different immunoassays. In order to investigate this phenomenom further, we have studied the effect of differences in assay kinetics within the same immunoradiometric assay (IRMA) configuration on LH measurement in sera from different endocrine states.

Three pairs of monoclonal/polyclonal two-site IRMA systems for LH were developed from three LH monoclonal antibodies and a common polyclonal anti-human chorionic gonadotrophin. For IRMA systems a short and long assay, which were different only with respect to the incubation time (1/2 h and overnight respectively), of the labelled monoclonal first antibody were performed. The IRMAs were all standardized against the LH international reference preparation 68/40. LH concentrations were measured by all the IRMAs in sera obtained from normal men ( n = 11) and from women with polycystic ovarian syndrome (PCO; n = 13). In normal men, there were no differences in LH estimates between the short and the long assays of the three IRMA systems, and the ratios of long to short assays were similar for all the systems. However, in PCO there were significant differences between short and long assays and the ratios of long to short assays were different for the IRMA systems.

These results indicate that kinetic differences between IRMAs of the same antibody configuration can be associated with differences in measured LH concentrations, depending on the endocrine status of the sera studied. As LH glycoform patterns are known to differ between normal men and PCO, the observed changes in LH estimates may be due to the different glycoform composition.
The Entero-Insular Axis in Polycystic Ovarian Syndrome
Authors: Garaa R , Norris F , Wright J , Morgan L , Hampton S , Watkins K , Marks V
Published: February 2014

We investigated the contributions made by the entero-insular axis, proinsulin and the fractional hepatic extraction of insulin to the hyperinsulinaemia characteristic of polycystic ovarian syndrome (PCOS). We measured plasma glucose, gastric inhibitory polypeptide (GIP), glucagon-like peptide-1 (7–36 amide) (GLP-17–36 amide), immunoreactive insulin (IRI), intact proinsulin (IPI), and C-peptide concentrations during a 75 g oral glucose tolerance test in seven normal weight women with PCOS and eight healthy women.

Women with PCOS had higher fasting ( P = 0·05) and integrated ( P < 0·01) IRI concentrations than controls. Fasting C-peptide levels were similar in both groups but integrated C-peptide ( P < 0·05) concentrations were greater in PCOS subjects than controls. Fasting and integrated concentrations of glucose, GIP and GLP-17–36 amide were similar in subjects with PCOS and controls. Although fasting IPI concentrations were similar in both groups, integrated IPI concentrations were higher ( P = 0·05) in patients with PCOS. Women with PCOS had similar fasting but higher ( P <0·05) integrated IRI: C-peptide molar ratios than controls. Fasting and integrated IPI: IRI molar ratios were similar in both groups.

These results confirm that lean women with PCOS have peripheral hyperinsulinaemia. The mild fasting hyperinsulinaemia is due to increased pancreatic secretion, whereas the stimulated hyperinsulinaemia is due to both pancreatic hypersecretion and reduced fractional hepatic extraction of insulin. Hyperproinsulinaemia is modest and appropriate in PCOS. GIP and GLP-17–36 amide do not contribute to the stimulated hyperinsulinaemia in PCOS.
Serum Osteocalcin, Total and Bone-Specific Alkaline Phosphatase following Isolated Tibial Shaft Fracture
Authors: Bowles Shirley A , Kurdy Nazeer , Davis Alison M , France Michael W , Marsh David R
Published: February 2014

We report the changes in osteocalcin concentrations and in total and bone specific alkaline phosphatase activity occurring in the twenty week period following tibial shaft fracture in 20 subjects.

Bone formation during the healing process is reflected by progressive increases in the concentration of osteocalcin and bone specific alkaline phosphatase after week 5 and the latter correlated with the height and weight of the subject. In the early post injury period, total alkaline phosphatase activity increased whereas that of the bone isoenzyme initially fell, starting to rise again during the second week. After an immediate post injury rise, osteocalcin concentration also decreased, reaching a nadir by week 5.

As only three of our subjects demonstrated delayed union, we have not been able to demonstrate that biochemical monitoring of the healing process can provide an indication of prognosis in tibial shaft fracture.
Measurement of Human Igf-II Using Sephacryl Extraction: A Rapid and Reliable Assay Method
Authors: Mason Barbara H , Tatnell Michele A , Holdaway Ian M
Published: February 2014

Measurement of insulin-like growth factor II (IGF-II) in human serum is complicated by the presence of IGF binding proteins and usually involves cumbersome extraction procedures followed by radioimmunoassay. We have utilized an extraction process developed for measuring insulin-like growth factor II in ovine serum using Sephacryl HR100, and have applied this to the extraction of human samples followed by radioimmunoassay for human IGF-II. The assay yielded 98% recovery of unlabelled IGF-II, parallelism between dilutions of eluate and the standard curve, complete removal of binding proteins and near-complete removal of IGF-I, and intra- and interassay coefficients of variation of 5% and 9%, respectively. The normal range for serum IGF-II in women was 490–1056 μg/L, and IGF-II levels were positively correlated with serum concentrations of insulin-like growth factor binding protein-3 (IGFBP-3) but not with IGF-I levels. Mean serum concentrations of IGF-II were reduced below normal in a number of hypopituitary patients and children with short stature and IGF-II concentrations in these subjects correlated positively with IGF-I and IGFBP-3. In acromegalic patients IGF-II levels were usually normal and were negatively correlated with IGF-I concentrations. From our experience with the above results the present assay appears particularly suitable for clinical measurements and research projects where high sample throughput is required.
Inhibition by Sodium Thiophosphate and L-P-Bromotetramisole of Neutrophil Alkaline Phosphatase in Normal and Trisomy 21 Pregnancies
Authors: Denier Colette J , Vergnes Henri A , Brisson-Lougarre Andrée , Grozdea Jean G , Fournie Alain E , Klaebe Alain G , Perie Jacques J
Published: February 2014

Alkaline phosphatase concentrations are known to increase in blood neutrophils of normal pregnant women. The main kinetic parameters of this enzyme were analysed and compared in a group of 30 women with normal pregnancies and a group of 11 women whose fetuses had trisomy 21 (Down's syndrome = DS). The subjects were studied at an identical stage of gestation. Significant changes occurred in thermal stability and urea resistance in cases of DS pregnancies. We also investigated the inactivation constants for two chemicals: L-p-bromotetramisole, an uncompetitive inhibitor, and sodium thiophosphate, a competitive inhibitor. Ki measured for the two inhibitors were found to be significantly lower in cases of pathological pregnancies. The patterns observed in inhibition constants extend the biochemical characteristics of the atypical isoenzyme expressed in neutrophils of women with DS pregnancies.
Steroid Sulphatase Deficiency: Identification of Heterozygotes Using Hydrolysis of Dehydroepiandrosterone Sulphate by Peripheral Leucocytes
Authors: Lowis E I , Oakey R E
Published: February 2014

Diagnosis of X-linked recessive ichthyosis, which is expressed only in males, can readily be made by measurement of leucocyte steroid sulphatase activity. However, because the gene for steroid sulphatase activity partly escapes from the process of X-chromosome inactivation associated with gene dosage compensation, identification of heterozygotes (females) is more difficult.

We have measured the steroid sulphatase (by hydrolysis of dehydroepiandrosterone sulphate) and β-glucuronidase (by hydrolysis of methylumbelliferyl glucuronide) activities in leucocytes from 18 heterozygotes, 100 normal females, 100 normal males and 11 affected subjects.

When the ratio of the activities of steroid sulphatase and β-glucuronidase in mixed leucocytes was plotted as a function of the steroid sulphatase activity, 85% heterozygotes were distinguished from normal females. Measurement of steroid sulphatase activity alone with these cells enabled identification of 78% heterozygotes. Measurements on mononuclear leucocytes were much less effective.

Thrombocytes showed 1% of the steroid sulphatase activity of leucocytes. In females, leucocyte steroid sulphatase activity was independent of the stage of the ovarian cycle at which the cells were collected.
Decision Support Techniques for the Interpretation of Quantitative Amino Acid Data
Authors: Briddon A
Published: February 2014

Quantitative amino acid profiles are traditionally reported as a list of individual concentrations and reference intervals. By the use of selected examples it is shown that alternative methods of presenting and assessing these results can enhance both the quantity and quality of information. A candidate decision support tool is offered as an aid to the interpretation of amino acid profiles.
Vitamin E and Exercise in the Northern Ireland Population
Authors: Duly E B , Trinick T R , Kennedy D G , Macauley D , McCrum E , Stott G , Evans A E , Sweeney K , Boreham C A G , Sharpe P
Published: February 2014

Serum vitamin E, vitamin E/cholesterol and physical activity and fitness were examined in a representative cross section ( n = 1600) of the Northern Ireland population as part of the Northern Ireland health and activity survey. Serum vitamin E levels were measured by high-performance liquid chromatography, cholesterol by an enzymatic method, physical activity profile was recorded by computer assisted interview and physical fitness was determined by estimation of VO2 max. The levels of serum vitamin E and vitamin E/cholesterol ratio in the Northern Irish population were similar or higher than in other populations with lower incidences of coronary heart disease. The assessment of activity showed that 75% of the population fell below recommended activity levels likely to confer a cardioprotective effect. A significant relationship ( P = 0·01) was found in males between serum vitamin E levels and lifetime participation in physical activity. Otherwise no relationship was found between serum vitamin E or vitamin E/cholesterol ratio and physical activity or fitness in the population.
Comparison of the Effect of Different Inhibitors on the Non-Enzymatic Glycation of Rat Tail Tendons and Bovine Serum Albumin
Authors: Menzel Ernst Johannes , Reihsner Roland
Published: February 2014

The biomechanical and biochemical properties of collagen are changed by non-enzymatic glycation culminating in increased cross-linking. We have previously shown that dibasic amino acids such as L-arginine inhibit in vitro the non-enzymatic glycation of soluble proteins and insoluble connective tissue macromolecules. In the present in vitro study we obtained evidence that the nucleophilic hydrazine derivative aminoguanidine and the non-steroidal antirheumatic drug ibuprofen inhibit the formation of fluorescent advanced glycation end products (AGEs) to a comparable extent, while arginine is ineffective as a consequence of its tendency to form AGEs itself. Periodic replacement of glycated arginine in the rat tail tendon system, however, engendered an inhibition of fluorescence similar to that obtained by the other inhibitors. Long-term glycation of rat tail tendons caused a significant increase in Young's modulus, which could also be inhibited by periodically renewed arginine. In contrast to ibuprofen, aminoguanidine and arginine-lysine inhibited the marked increase in maximum contraction force of long-term glycated rat tail tendons. As opposed to other inhibitors, aminoguanidine also reduced the thermal contraction force of native tendons, shifted the maximum contraction temperature to markedly lower values and solubilized a significant part of the rat tail tendon collagen. These findings indicate that the in vitro alterations of rat tail tendon collagen induced by non-enzymatic glycation can be prevented by arginine, arginine-lysine and aminoguanidine. However, collagen structure is seriously affected by aminoguanidine.
Plasma Histamine in Patients with Tropical Disorders and Acute Asthma Who Exhibit Eosinophilia
Authors: Bruce Christine , Taylor WH
Published: February 2014

The mean venous plasma histamine of 14 patients with eosinophilia was 18·7 nmol/L (range 0–86·4) and significantly higher than that of 4·0 nmol/L for 29 normal subjects [range 0–10·8 ( P < 0·01)]. The eosinophilia of schistosomiasis (two patients), strongyloidiasis (one patient), and asthma (four patients) was accompanied by raised plasma histamine concentrations. Normal levels occurred in onchocerciasis (four patients) and in two of three patients with non-specific eosinophilia. Plasma histamine concentrations in the tropical disorders are not thought to have been reported previously. The initial arterial and venous plasma histamine concentrations of 14 acute asthmatics, were significantly positively correlated with the respective arterial ( P < 0.01) and venous ( P < 0.05) eosinophil counts. The results in acute asthma support the hypothesis that increased plasma histamine may have a role in promoting eosinophil release from the bone-marrow. Eosinophilia is not dependent upon a raised plasma histamine in some disorders, but may be in others, such as schistosomiasis.
The Effect of Eccentric Exercise on Serum Creatine Kinase Activity in Different Ethnic Groups
Authors: Sherwood R A , Lambert A , Newham D J , Wassif W S , Peters T J
Published: February 2014

Eccentric exercise causes release of muscle creatine kinase (CK) 3–4 days after exercise. Racial variation in basal serum CK has been reported but the reasons for this are unknown. We studied 30 subjects of different ethnic origin (Caucasian, Afro-Caribbean, Asian) before and after eccentric exercise. Basal serum CK was significantly higher in the Afro-Caribbean group (201 ± 134IU/L, median ± SD) compared to Caucasians (81 ± 57 IU/L). The Asian group had intermediate CK values (144 ± 93 IU/L).

The intra-individual range of peak post-exercise CK values obtained was very wide (95–30 200 IU/L) with little difference in median CK between the Afro-Caribbean (8450 ± 9020 IU/L) and Caucasian groups (7600 ± 8800 IU/L). The median value for the Asian group was lower (594 ± 5410 IU/L). A sub-group of 15 individuals undertook a second bout of exercise 2 weeks later and all subjects demonstrated a training effect resulting in a marked attenuation of enzyme efflux. The variation in CK between the ethnic groups was not related to measurements of muscle strength or body mass, although the torque: body mass ratio followed the same order as the basal CK, i.e. the Afro-Caribbeans had the highest values. These results highlight the importance of considering ethnic origin and previous exercise history when interpreting serum CK assay results.
Four Radioisotopic Immunoassays of Free Thyroxine Compared
Authors: Van Blerk M , Smitz J , Rozenski E , Mees M , Roelandt P , Laermans L , Callewaert M , Van Steirteghem A C
Published: February 2014

The analytical and clinical performance of three radioisotopic methods for the measurement of free thyroxine—GammaCoat™ (125I) Free T4, RIA-gnost® FT4 and Amerlex-MAB* FT4—were assessed in comparison with the Sclavo two-step chromatographic method. The analytical evaluation indicated excellent performance of Amerlex-MAB FT4. For the other methods, precision and sensitivity were comparable and acceptable for routine use. Only GammaCoat FT4 showed a significant positive intra-assay drift. None of the methods showed a significant correlation with thyroxine-binding globulin. Amerlex-MAB FT4 results were weakly positively correlated with albumin. Sclavo, RIA-gnost and particularly GammaCoat FT4 results were affected by the presence of increased concentrations of free fatty acids. All methods classified hyperthyroid patients correctly. Slight overlaps existed between the hypothyroid and euthyroid populations. Significant decreases in free thyroxine (FT4) during the third trimester of pregnancy were detected by all assays. In patients with chronic renal failure, serum FT4 was within reference limits more often when measured by Sclavo or Amerlex-MAB methods than by RIA-gnost or GammaCoat techniques. In conclusion, all assays performed well from both technical and diagnostic points of view.
A Simple HPLC Method for the Separation of Amphetamine Isomers in Urine and its Application in Differentiating between ‘Street’ Amphetamine and Prescribed D-Amphetamine
Authors: Palfrey S , Labib M
Published: February 2014

D-amphetamine has been increasingly prescribed to treat amphetamine abusers. Prescribing D-amphetamine requires laboratory evidence or confirmation of current use of ‘street’ amphetamine, using a method which should be capable of differentiating between ‘street’ amphetamine and prescribed D-amphetamine. We have developed a simple high-performance liquid chromagraphy (HPLC) method for the separation of the two isomers of amphetamine in urine and have assessed its use in differentiating between ‘street’ amphetamine and prescribed D-amphetamine. The method is reproducible, free from interference and has a detection limit of 0·1 μg/mL for each isomer. Urine from patients prescribed D-amphetamine contained only a trace amount of L-amphetamine (less than 4%) whereas urine from those taking ‘street’ amphetamine contained more than 50% L-amphetamine. The method is applicable to confirmation of ‘street’ amphetamine misuse and for monitoring patient compliance with treatment. The presence of 4% or less L-amphetamine in urine would suggest that the patient is only taking prescribed D-amphetamine whereas the presence of L-amphetamine in higher concentrations suggests that the patient is taking ‘street’ amphetamine, with or without prescribed D-amphetamine.
Familial Defective Apolipoprotein B-100: A Study of Patients from Lipid Clinics in Scotland and Wales
Authors: Wenham Philip R , Bloomfield Peter , Blundell Gillian , Penney Michael D , Rae Peter W H , Walker Simon W
Published: February 2014

Familial defective apolipoprotein (apo) B-100 (FDB) is an autosomal codominant disorder, which may be associated with hypercholesterolaemia. The defect is caused by the substitution of glutamine for arginine at amino acid residue 3500 of apo B-100. A total of 357 hypercholesterolaemic patients, 48 with a clinical diagnosis of familial hypercholesterolaemia attending lipid clinics in Scotland and Wales, were screened for the presence of FDB. Seven unrelated individuals, five of whom had a family history of coronary heart disease, and a further 11 first-degree relatives, were shown to be heterozygous for the mutation. Pedigree analysis demonstrated the mutation to be present on a single haplotype, suggesting that in Britain it is inherited from a common ancestor. Treatment of 11 heterozygous individuals with lipid-lowering medication showed falls in total and low density lipoprotein cholesterol ranging from 11·6 to 38·8% and 5·3 to 49·5%, respectively. In view of the condition's association with coronary heart disease and hypercholesterolaemia, it may be worthwhile identifying carriers attending lipid clinics, so that affected siblings can be offered cholesterol-lowering treatment where necessary.
The Effects of Cigarette Smoking and Alcohol Consumption on Serum Liver Enzyme Activities: A Dose-Related Study in Men
Authors: Whitehead T P , Robinson D , Allaway S L
Published: February 2014

The separate and joint effects of cigarette smoking and alcohol consumption on serum activities of the enzymes aspartate aminotransferase (AST), alanine aminotransferase (ALT) and γ-glutamyl transferase (GGT) were investigated in 46775 men attending the BUPA Health Screening Centre in London during the period 1983–1987, after allowing for differences in age, body mass index and exercise level. As expected, all showed a significant positive correlation with alcohol consumption. Cigarette smoking produced a significant increase in GGT activity in all drinking categories bar teetotallers, particularly for those smoking more than 20 cigarettes per day. There were no changes of clinical significance in AST or ALT activities with smoking. We postulate that the combined effects of alcohol and smoking on GGT activity are a result of induction of the enzyme by both alcohol and nicotine. Smoking as well as drinking habits should be taken into account when assessing the significance of an individual's enzyme activities.
Patients with Recurrent Renal Stones Have a Physico-Chemically Altered Urinary Tamm-Horsfall Glycoprotein Profile
Authors: Jefferson A , Reynolds T M , Elves A , Wierzbicki A S
Published: February 2014

We describe an immunoblotting method for examining the electrophoretic properties of urinary Tamm-Horsfall protein. Using this method we tested frozen urine samples from a group of 37 thoroughly investigated recurrent idiopathic renal calcium stone formers and compared this with fresh urine from 19 non-stone forming laboratory staff. We found that there was a statistically significant different pattern of Tamm-Horsfall protein bands in the two sets of urines, with stone formers tending to have two bands and non-stone formers tending to have three bands. This could have been due to storage artefact and therefore a further group of 13 fresh urines from unselected renal stone formers was tested. A smaller proportion of these cases showed the two-band pattern, possibly because not all of this group were idiopathic calcium stone formers. This suggests that but does not prove that there is no significant storage artefact and that there may be an in vivo effect causing stone formation.
Variability of the Methionine Loading Test: No Effect of a Low Protein Diet
Authors: den Heijer Martin , Bos Gerard M J , Brouwer Ingeborg A , Gerrits Wim B J , Blom Henk J
Published: February 2014

The methionine loading test is used for the diagnosis of impaired methionine/homocysteine metabolism, in particular the transsulfuration pathway. Usually this test is performed on a low protein diet to control the intake of methionine. However, this is inconvenient and relatively expensive. In this study we compared the effects of a low protein diet and a standard diet on methionine loading test in 28 subjects (crossover design). The mean difference in homocysteine concentration after methionine loading between the two diets was 1.3 [confidence interval (CI) 95%-1.0–3.6]μmol/L which demonstrates that a special low protein diet is not essential in the performance of the methionine loading test. We also observed that 3 weeks after the first methionine loading test, fasting serum concentration of folate was higher and vitamin B12 concentration was lower.
The Effects of Gestation Dating on the Calculation of Patient-Specific Risks in Down's Syndrome Screening: Multivariate Case
Authors: Bishop J C , Dunstan F D J , Nix A B J
Published: February 2014

When screening for Down's syndrome using biochemical markers, the measurements are adjusted for the gestational age of the fetus because the concentrations of the markers are known to change with gestational age. This adjustment is performed by referring each marker measurement to the population median for that marker for the appropriate estimated gestational age group. The measurement of gestational age is subject to error, whichever method is used, and so the population median used is usually the median of a mixture of distributions for different true gestational ages. Most screening programmes aim for a specific number of weeks and this produces a concentrated distribution of true gestational ages. This fact, combined with dating errors, leads to an asymmetric mixture for each gestational age group and hence to bias in the estimates of the medians. In a previous communication we have shown how the proportions in this mixture distribution can be estimated and how the true medians corresponding to a true gestational age can be estimated. The calculations presented were performed using a single marker, and the details of our method were restricted to this situation. This paper extends the method to the multimarker situation and, as expected, leads to a gain in the detection rate for a specified false positive rate. The true patient-specific risk estimates are again markedly different from the quoted nominal value obtained by ignoring the dating errors. The data set on which the method is illustrated uses two markers, although the technique generalises in an obvious way to more than two.
Benzodiazepine Misuse by Drug Addicts
Authors: Garretty Deborah J , Wolff Kim , Hay Alastair W M , Raistrick Duncan
Published: February 2014

Using a high-performance liquid chromatography method, we measured seven commonly prescribed benzodiazepines (chlordiazepoxide, nitrazepam, nordiazepam, oxazepam, lorazepam, temazepam and diazepam) in 100 urine samples obtained from patients attending the Leeds Addiction Unit. All of the urines selected for investigation were positive for benzodiazepines using an EMIT (Enzyme Immunoassay) screen. Forty-four of the urines contained a range of benzodiazepines, none of which had been prescribed.

Nitrazepam was detected most frequently (61 urine samples), but had not been prescribed to any of the patients in this study. Chlordiazepoxide was detected in 49 urine samples, although it had been prescribed to only five patients. Temazepam was detected in 28 urine samples. Fourteen patients providing 21 urine samples had been prescribed temazepam for treatment. However, temazepam was detected in only 14 of these samples. Multiple benzodiazepine abuse was evident from the high rate of detection of unrelated benzodiazepines.
Serum Total Antioxidant Activity after Myocardial Infarction
Authors: Miller Nicholas J , Johnston John D , Collis Clifford S , Rice-Evans Catherine
Published: February 2014

Serum total antioxidant activity (TAA), albumin and uric acid were measured on admission, and for the next 2 days in 56 patients suffering myocardial infarction, 20 of whom received streptokinase. The ‘antioxidant gap’, the difference between the serum TAA and the sum of the serum albumin and uric acid activity, was calculated. No significant changes in serum total antioxidant activity were observed in either group of patients between admission, day 1 and day 2. However, a decline in the ‘antioxidant gap’ after myocardial infarction was associated with a significantly higher mortality.
Ovarian Steroids Regulate the Expression of Basic Fibroblast Growth Factor and its mRNA in Fibroblasts Derived from Uterine Endometrium
Authors: Fujimoto Jiro , Hori Masashi , Ichigo Satoshi , Tamaya Teruhiko
Published: February 2014

The role of stromal cells in basic fibroblast growth factor (FGF) supply for endometrial neovascularization during the menstrual cycle was investigated. The concentrations of intracellular and secreted FGF, and FGF mRNA expression were determined in fibroblasts derived from uterine endometrium as a substitute for stromal cells. The influence of sex steroids on protein and mRNA expression was investigated.

The concentration of FGF and its mRNA expression in the fibroblasts was significantly increased by oestradiol, and these increased concentrations were diminished by progesterone. It is suggested that oestrogen stimulates FGF secretion from the stromal cells, an effect which is inhibited by progesterone. Therefore, endometrial neovascularization might be partially regulated by stromal-derived FGF under the influence of sex steroids, through a paracrine cell-to-cell interaction.