Monitoring ovarian function in marmosets and tamarins by the measurement of urinary estrogen metabolites

Practical aspects of urinary estrogen analysis were considered with regard to establishing simple and reliable methods for monitoring ovarian function in marmosets and tamarins. Changes in the hormone:creatinine ratio in small volumes of urine from the common marmoset were significantly correlated with changes in 24-h excretion. Comparison of the metabolism and excretion of estrogens during the ovarian cycle in the common marmoset and cottontop tamarin revealed interesting species differences. High concentrations of conjugated estrone were measured in marmoset plasma, but estradiol 17β was the predominant estrogen in urine. In contrast, estrone was the most abundant estrogen measured in tamarin urine. Both species excreted very little estriol. Sulfates and glucuronides were present in urine in similar proportions before ovulation in the marmoset, although after ovulation sulfates were the more abundant. Conversely, most of the estrogens in tamarin urine appeared to be conjugated as glucuronides. Direct assay for estrone sulfate was applied to the measurement of urinary estrogen excretion during the ovarian cycle in a marmoset. The results compared well with those for total estradiol 17β after hydrolysis and ether extraction. The use of direct assays for conjugated estrogens in small volumes of urine is suggested as a practical method for monitoring ovarian function in marmosets and tamarins.     

Determination of urinary arsenic, mercury, and selenium in steel production workers

The fumes and dust of trace elements and their compounds are very toxic and have been related to an increase in the incidence of diseases. Occupational exposure to toxic metals and metalloids can be determined by means of workplace air measurements and biological monitoring. The aim of our investigation was to determine the concentrations of As, Hg, and Se in urine samples under routine clinical laboratory conditions. To assess the reliability of these methods, critical factors such as detection limit(s), calibration range(s), cost, accuracy, and precision were studied. The method was employed for the quantitative determination of arsenic, mercury, and selenium in urine samples from steel production and quality control workers and healthy unexposed controls. After pretreatment with acids, the samples were digested by means of a microwave oven. Arsenic was determined by hydride atomic absorption spectrometry and mercury was determined by cold vapor atomic absorption spectrometry, whereas selenium was determined by a graphite furnace atomic absorption spectrometry. The results indicate those urinary arsenic, mercury, and selenium levels of the exposed workers are significantly higher than those of the controls. The possibility that these elements are involved in the etiology of diseases is discussed and recommendations are made to improve workplace ventilation and industrial hygiene practices.     

2-Acetamidoglucal,a new metabolite isolated from the urine of a patient with sialuria

A new metabolite, namely 2-acetamidoglucal, has been found in the urine of a patient with sialuria in addition to the metabolites N-acetylneuraminic acid, N-acetylmannosamine, N-acetylglucosamine and N-deoxy-2,3-dehydro-Nacetylneuraminic acid reported earlier. The structure has been identified by mass spectrometry and 360 MHz proton nuclear magnetic resonance spectroscopy and verified by synthesis. All accumulated compounds fit into the metabolic pathway for the biosynthesis of CMP-N-acetylneuraminic acid. Sialuria is discussed in terms of a failure of regulation of UDP-N-acetyl-glucosamine 2-epimerase.     

Determination of iodide amounts in urine and water by isotope dilution analysis

Urinary iodide and iodine in drinking water were determined in 318 healthy children aged 0 to 18 yr living in Izmir and environmental rural and urban areas in the western part of Turkey. The method is based on substochiometric isotope dilution analysis. Iodide was precipitated by substoichiometric amounts of AgNO₃. Iodide-131 was used as a tracer. Electrophoresis was performed to separate Ag¹³¹I from excess¹³¹I-. The Ag¹³¹I zone was cut off the electrophoresis paper and counted with a Nal(Tl) scintillation counter. Count rates were plotted versus added KI concentrations. The unknown iodide amount was found by using these linear plots. Iodide concentration ranges were within 1.8 –100.45 Μg/L in the analyzed drinking water samples. The mean value was 44.14 ±17.33 Μg/L and the median was 58.08 Μg/L. Urinary iodide concentration ranges were 0.22 –142.22 Μg/L. The median of the distribution was 37.71 Μg/L and the mean was 40.30 ±24.05 Μg/L. The results show that the examined area suffers moderate iodine deficiency.     

Highly sensitive spectrofluorimetric method for rapid determination of orciprenaline in biological fluids and pharmaceuticals

Orciprenaline sulphate (ORP) is a direct‐acting sympathomimetic with mainly beta‐adrenoceptor stimulant activity. It is used as a bronchodilator in the management of reversible airway obstruction. For the first time, a rapid highly sensitive spectrofluorimetric method is described that is relied on measuring the fluorescence spectra of ORP at acidic pH and without addition of any chemical reagents. The relative fluorescence intensity was measured at 310 nm and after excitation at 224 nm. ORP native fluorescence was calibrated in both water and acetonitrile as diluting solvents. The method was designed to estimate the drug in miscellaneous matrices with high accuracy and precision. Linear ranges of calibration curves were 30.0–400.0 ng/ml and 10.0–240.0 ng/ml in water and acetonitrile, respectively. The detection limits were calculated and reached as low as 3.3 and 3.1 ng/ml, respectively, representing the ultra‐sensitivity of the proposed method. This result permitted application of this method for spiked human plasma and urine and was used as a preliminary investigation with good percentage recovery (89.4–106.8%). The application was further extended to analyse ORP in its pharmaceutical formulations. The method was validated in compliance with International Council of Harmonization (ICH) Guidelines.     

Evaluation of Urinary Protein Precipitation Protocols for the Multidisciplinary Approach to the Study of Chronic Pelvic Pain Research Network

Standardization of sample collection, shipping, and storage has been a major focus of biorepositories servicing large, multi-institute studies. The standardization of total protein concentration measurements may also provide an important metric for characterizing biospecimens. The measurement of total protein concentration in urine is challenging because of widely variable sample dilutions obtained in the clinic and the lack of a reference matrix for use with a standard curve and blank subtraction. Urinary proteins are therefore typically precipitated and reconstituted in a reference solution before quantitation. We have tested three different methods for protein precipitation and evaluated them using variability in total protein concentration measurement as a metric. The methods were tested on four urine samples ranging from very concentrated to very dilute. A method using a commercially available kit provided the most reproducible results, with average coefficients of variation <10%. Addition of a freeze/thaw did not lead to significant protein loss or additional variability. Samples were titrated and the measurements obtained appeared to be linearly correlated with sample starting volume. This method was applied to analysis of 77 urine biorepository samples and provided reproducible results when the same sample was assayed on different microwell plates.     

Including oxidisation of ammonia in the eutrophication impact category

Oxygen depletion of lake and seawater is a serious condition with large implications for biodiversity. Therefore, in LCA, the potential oxygen demand of water emissions is estimated under the label eutrophication impact category. This impact category should contain the impact of water emissions on the total oxygen consumption in the receiving water. This means that it should include both primary and secondary oxygen consumption. In spite of this, the oxygen needed to oxidise ammonia has normally not been taken into account when quantifying the eutrophication impact category. In this paper, weighting factors for ammonium/ammonia are suggested for the eutrophication impact category. It is shown that, for treated wastewater, the amount of oxygen needed for nitrification of ammonia is important when compared to the potential eutrophication calculated using the current recommended weighting factors. These weighting factors take into account oxygen needed to oxidise the organic matter in the wastewater emission and that needed to degrade the algae potentially grown due to the emission of nutrients.     

Diet treatment of branched chain ketoaciduria studied by proton magnetic resonance spectroscopy

Summary A novel nuclear magnetic resonance method is proposed for the diagnosis and follow-up of patients affected by branched chain ketoaciduria. The method allows quantitation of the branched chain amino acids (BCAA's) such as leucine, isoleucine and valine and of related keto- and hydroxy acids by means of a single spectrum. The method implies short time of analysis, as opposed to the very long time required by the techniques currently in use (amino acid analyzer combined with gaschromatography/mass spectrometry of keto- and hydroxyacids), it is easy and suitable for adjustements of the dietary treatment even on a daily basis. The case of a 15 days old newborn child, presenting muscular hypertonicity was unambiguously diagnosed in few minutes by means of one single NMR spectrum of urine. More interestingly, NMR spectra of serum in the following days were suitable for quantitating amino-, and keto acids as well as other metabolites of relevance in the follow up of the dietary treatment of the disease. After a diet lacking of BCAA's, to eliminate keto acids, a low BCAA diet was introduced, that succeeded in keeping the serum levels of the three amino acids within the normal range, while dropping the related keto acids.