Screening complex biological specimens such as exhaled air, tissue, blood and urine to identify biomarkers in different forms of cancer has become increasingly popular over the last decade, mainly due to new instruments and improved bioinformatics. However, despite some progress, the identification of biomarkers has shown to be a difficult task with few new biomarkers (excluding recent genetic markers) being considered for introduction to clinical analysis. This review describes recent advances in gas chromatographic methods for the identification of biomarkers in the detection, diagnosis and treatment of cancer. It presents a general overview of cancer metabolism, the current biomarkers used for cancer diagnosis and treatment, a background to metabolic changes in tumors, an overview of current GC methods, and collectively presents the scope and outlook of GC methods in oncology.

Urinary steroids have been studied during early and late pregnancy in domestic horses or sporadic samples at various stages of pregnancy in wild equidae. In our studies, urinary estrone sulfate (E1S) and pregnanediol glucuronide (PdG) were monitored throughout pregnancy in six pony mares by enzyme immunoassay (EIA). Both hormones were corrected by creatinine (Cr) index to compensate for the variation in specific gravity. The mean concentration for ElS, (μg/mg Cr), was .38 ± .03 at d 0, decreased to .17 ± .04 at d 1, and maintained at less than .5 μg/mg Cr until d 30. Although, there was an apparent increase to .80 ± .34 at d 34 (NS, P = .122), the first significant increase was .69 ± .15 at d 46 (P = .0275). Mean concentrations remained relatively stable at this approximate level until d 60. This level was followed by a sustained significant increase observed from d 60 onwards. Mean concentrations of El S increased to 1.11 ± .25, 2.01 ± .45, and 5.48 ± 1.47 at d 64, d 76, and d 86, respectively. Levels of EIS further increased reaching a peak of 143.3 ± 9.51 at d 142 (P = .0006), with maximum for individual mares ranging from d 114 to 170, and also ranging from 115.4 to 286.1 pg/mg. In all cases, maximum concentrations were followed by a gradual decline toward parturition with a more rapid decrease 1 to 3 days before parturition. The first significant decrease following the maximum concentration was 91.40 ± 13.11 (P = .0024) at d 184. Estrone sulfate was 12.1 ± 3.8 one day prepartum and decreased to .4 ± .1 and .1 ± .01 at d 1 and 4 postpartum, respectively. The mean concentrations of PdG (ng/mg Cr) increased from 147 ± 4.3 at d 0 to 50.87 ± .17 (NS, P> .05), 36.8 ± 8.1 (P = .016), and 27.6 ± 7.3 (P = .049) at d 6, 8 and 10, respectively. This increase was followed by a decline and generally the levels fluctuated ranging from 20 to 30 ng/mg Cr until d 80. At d 86, the PdG levels increased to 54.7 ± 11.7 (P = .033). This was followed by a further increase to 141.8 ± 21.4 (P = .0139, compared to d 93) at d 135, then continued to increase to 213.0 ± 25.2 at d 198, and remained at this approximate level until d 303. During the last month of gestation, the mean concentrations of PdG increased from 171.8 ± 9.8 at d 29 prepartum to reach a peak of 388.4 ± 108.6 at d 7 prepartum. Maximum concentrations were followed by a slight decrease to 354.5 ± 84.0 at d 1 prepartum and then decreased to 150.6 ± 23.4 and 39.6 ± 9.3 ng/mg Cr at d 1 and 4 postpartum. In comparing the two hormones, E1S remained baseline followed by a slight increase at d 35, whereas PdG was relatively stable until both hormones increased after d 70 of gestation. This might be related to secretion of both hormones by the fetus and their rapid metabolism by placenta. Estrone sulfate reached a peak at approximately d 142 followed by a decline toward parturition while PdG showed a rapid increase from d 70 to 150, followed by a slow sustained increase to d 300 then increased dramatically again before parturition, while El S continued to decline. The profile of these urinary hormones throughout pregnancy appeared to parallel previously published concentrations in blood. Since the patterns of urinary EIS and PdG are different, their sites and mechanism of metabolism are likely different. The results indicate that the presence of the feto-placental unit is important for the secretion of both estrogens and progestins throughout pregnancy and thus could be utilized as a reliable method for pregnancy determination after three months of pregnancy. / Graduation date: 1993

Odor signals mediate a variety of behaviors in animals across a diversity of taxa. Despite dramatic morphological differences between animals from different taxa, several important features of olfactory system organization and processing are similar across animals. Because of this similarity, a number of different organisms including mammals, insects, and decapod crustaceans serve as valuable model systems for understanding general principles of olfactory processing. As in other organisms, including both vertebrates and insects, the chemosensory system of decapod crustaceans is organized into multiple anatomically distinct neuronal pathways. The two main pathways (the aesthetasc/ olfactory lobe pathway and non-aesthetasc/ lateral antennular neuropil pathway) originate in different populations of antennular sensilla and project to different neuropils in the brain. The functional significance of this parallel organization is not well understood in crustaceans or in many other species. Although in some insect species the functions of parallel pathways are clearly delineated by the types of odors processed by each, functional differences between parallel pathways in other organisms are much less distinct. A critical step towards understanding the functional significance of the multiple chemosensory pathways is to identify the specific behaviors that are driven by each pathway. Using spiny lobsters and crayfish as model organisms, the importance of each pathway was examined in three different behavioral contexts: (1) orientation to a distant food odor, (2) shelter selection in response to conspecific chemical signals, and (3) determination of conspecific social status. In each study, selective ablations of specific populations of antennular sensilla were performed, and the behavior of ablated animals was compared to that of intact controls. Results show that either the aesthetasc or non-aesthetasc pathway is capable of driving orientation to food odors, suggesting functional redundancy between the pathways in this behavior. In contrast social odors are processed preferentially by the aesthetasc pathway rather than the non-aesthetasc pathway, suggesting a unique role for the aesthetasc pathway in this context. As in other organisms possessing multiple chemosensory pathways, the dual antennular pathways in crustaceans display both unique and overlapping functions depending on the chemicals examined, and the behavioral context in which the signal is presented.

One of today’s most widely spread health problems is urinary incontinence, affecting 60-80% of the US population from age 15 and up. Treatment based on the possibility to implant a scaffold seeded with the patients’ own urine-derived stem cells, hUSC, to regenerate the damaged muscle tissue, would prove effective. A main challenge in regenerating new tissue from cell-seeded scaffolds is the limited cell survival due to insufficient oxygen diffusion to the center of the scaffold. Ways of enhancing cell survival, and thereby, proliferation and differentiation, is by hypoxic preconditioning of the cells or implantation in an oxygen-release material. Hypoxic preconditioning has shown to enhance proliferation as well as the expression of vascular endothelial growth factor, VEGF, in for example human bone marrow derived stem cells, hBMSC. VEGF is involved in the establishment of vasculature structures and an upregulation of its expression may therefore help promote quicker angeogenisis, increasing the oxygen supply and the cell survival. Oxygen-release materials have shown to enhance cell survival and growth both in vitro and in vivo. This study aims to investigate the effect of hypoxia on hUSC, during 9 days of hypoxic culturing (2.0% ± 0.1% O2) with and without oxygen-release material (PLGA 75:25 with 5 w% CPO) in vitro. hBMSC, and human smooth muscle cells, hSMC, have been used as control groups. Cell proliferation, morphology, differentiation, production of VEGF, and expression of hypoxia inducible factor HIF-1α have been studied. According to the results, combining hypoxic preconditioning of hUSC with implantation in oxygen-release material could be an effective way to regenerate muscular tissue. Hypoxic preconditioning enhanced cell proliferation, production of VEGF, and HIF-1α expression. The increase of VEGF and HIF-1α would promote vascularization when implanted. The oxygen-release material showed possible promotion of cell differentiation, which would augment the hUSCs’ myogenic differentiation, while supplying oxygen until the tissue’s vascular structure has been established.

From the analysis of the Dongsheng project which applied ecological sanitation in multi-storey buildings in China, we found that technical deficiencies, managerial problems and incorrect usage of the urine-diverting toilets resulted in the poor performance of the Dongsheng ecological sanitation system. Lack of standards or guidelines, and lack of policies or regulations are significant challenges in implementing the ecological sanitation system. Residents in the Dongsheng eco-town have positive attitudes toward reusing human faeces and urine in agriculture since China has a long tradition of this. However, the residents did not know very well about the value of urine, and the local farmers did not want urine. Women in China are more concerned with sanitation from a health aspect than men. Higher education level is connected to more positive attitudes towards human faeces and urine and their reuse. However, there is no link between income status and the attitudes of users. The supply chain for ecological sanitation systems is weak in urban China and needs to be improved. Apart from the demand side and supply side of ecological sanitation, enabling ecological sanitationto go to scale in an urban context depends on physical, environmental, technical and policy factors.

Ammonia is a nuisance gas in many swine barns. The overall objective of this research project was to model ammonia formation and transmission processes in a grower-finisher swine barn, by first modelling the ammonia production and emission from urine puddles on the floor surface and the ammonia emission from the slurry pit, and then incorporating these emission rates in a dynamic model that separates the room and slurry pit headspace as two separate, but linked, control volumes. A series of studies were conducted to gather more information about the processes affecting the ammonia emission rate from the floor surface and the slurry that were later included in the overall room model developed. The model was then used to investigate ammonia reducing techniques and technologies based on the understanding of ammonia production and transmission incorporated in the model. The first step in modelling the ammonia emission rate from the floor surface was to determine the frequency of urinations by grower-finisher pigs. Male and female pigs were observed three times during their finishing phase to determine their urination frequency over the course of a day. The average measured urination frequency was 0.62 ± 0.11 urinations pig-1 h-1. A sinusoidal dromedary model was developed to describe the daily variation in urination frequency for male and female pigs between 51 and 78 kg.<p>In order for the deposited urinations on the floor surface to emit ammonia, the urea in the urine must first be converted to ammonia and the urease enzyme catalyzes this reaction. Two methods, a fixed-time-point method using the indophenol assay for ammonium-nitrogen analysis and a continuous method using the coupled enzyme assay, were used to measure enzyme activity at the floor surface of a swine barn and were compared to reported urease activity levels in the literature. Using both methods, there appeared to be an ammonia-producing site on the floor surface or within the collected samples that made accurate measurements of urease activity impossible. A review of urease activity levels in the literature from dairy-cow houses suggest that urease activity will be lowest following floor-cleaning and increase quickly following fouling of the floor surface. Based on the literature review, a urease activity value of 5 g NH¬3 m-2 h-1 was suggested for use in ammonia emission modelling of fouled floor surfaces in swine barns until better measurements become available. <p>The ammonia emissions from 36 simulated urine puddles under a variety of temperature, air velocity and initial urea concentration conditions were measured in a bench-scale experimental set-up. The measurements were used to calibrate and validate a dynamic, mechanistic, urine puddle emission model that considered the processes of evaporation, urea conversion, change in liquid concentration and puddle pH in order to simulate the amount of ammonia emitted from a puddle. Based on the correlation coefficients (R) between measured and simulated values for water volume (R=0.99), total ammoniacal nitrogen concentration (R=0.90), and total emission (R=1.00), along with five other statistical tests for each simulated variable, the model was deemed accurate. The measurements and simulations in this experiment showed the impact of puddle pH, urease activity and changing environmental conditions on the average puddle emission rate. Puddle emission continued to occur as long as there was still water.<p> The impact of different slurry compositions on the ammonia emission rate from slurry pits was tested in another bench-scale experimental set-up with emission chambers. The emission chamber concentration data collected was used to calibrate and validate a developed slurry emission model. The collected slurry samples were concentrated mixtures of urine and feces from individually-housed animals fed different diets. An empirical equation was developed to express the amount of total ammoniacal nitrogen in the slurry that was in the form of ammonia (f) and thus volatile to the surroundings. Based on the empirical equation, the simulated value of f was between 0.03 and 0.08 and did not show the sensitivity to slurry pH that has been reported by other authors. The slurry emission model with the empirical equation for f was validated with ammonia emission measurements from eight different slurry samples and simulated hourly concentration measurements within 17% and five-day average concentration measurements within 3%. Further testing was recommended to ensure the model developed for concentrated manure in this study was applicable to the more dilute slurry found in swine barns. <p>Using the information gained in the previous experiments, a mechanistic model describing the dynamic ammonia concentration in the room and in the slurry channel headspace of grower-finisher swine barns, as well as the ammonia emitted to the surrounding environment was developed. Data was collected from two grower-finisher rooms to use as input data to the model and for calibration and validation purposes. The model calibration procedure determined that the amount of emissions originating from the slurry for the simulated room conditions was generally less than 5% of the total room emissions, the air exchange rate through the slatted floor was approximately 4% of the room ventilation rate, and that in the first two weeks of animal activity in a room the urease activity at the floor surface will increase. The model was validated using separate data from that used in the calibration process. The model simulated hourly room concentration levels within 2.2 ppm and 3-day average concentration levels within 1.6 ppm. The model simulations were more accurate for one room that was fed a typical grower-finisher diet compared to another room fed an experimental diet with lower protein content and sugar-beet pulp inclusion. <p>The dynamic model was tested for its sensitivity to various input factors in terms of the floor emission rate, slurry emission rate and total emission rate. An interesting aspect of the simulations was that increases in either floor or surface emission rate were compensated to a small extent by decreases in the other emission rate as a result of a reduced concentration gradient for mass transfer. The ammonia emission rate from the floor was most sensitive to changes in urease activity, fouled floor area and puddle area. The ammonia emission rate from slurry was most sensitive to changes in slurry pH. The impact of input variables on the total emission rate was dependant on the simulated proportion of the total ammonia emission coming from either the floor surface or slurry channel. Three ammonia reduction techniques were tested and evaluated on their impact to the total ammonia emission rate from a room compared to a given set of control conditions.<p>The work in this thesis highlighted the importance of ammonia emission from the floor surface. The proportion of ammonia originating from the slurry and from the floor surface respectively will vary on the specific conditions within the barn, and will impact the effect of any ammonia mitigation technique that is investigated or used.

Cystatin C, a 13 kDa low molecular weight protein is an inhibitor of cysteine proteases. Due to its low molecular weight and positive charge at physiological pH, it is freely filtered by the glomerulus and catabolized after reabsorption by proximal tubular cells with a low concentration (0.03-0.3 mg/L) in urine amongst healthy subjects. Urinary cystatin C is a potential biomarker detection of acute kidney injury (AKI) in the acute phase when patients are submitted to the intensive care unit. The aim in this report was to perform a full method validation of urinary analysis of cystatin C on a high throughput chemical analyzer by particle-enhanced turbidimetric immunoassay (PETIA) at the University Hospital in Uppsala, Sweden. The antigen excess, linearity, lower limit of quantification (LoQ), recovery, assay precision, stability and interference caused by haemoglobin was evaluated. No hook effect was observed, the assay was linear over the studied interval &lt;0.001-0.950 mg/L with a regression of R2=0.9994. The LoQ was calculated to 0.020 mg/L with a coefficient of variation (CV) ≤10% which was considered acceptable. The assay had a recovery between 93-100% and the assay precision had a total CV &lt;3.5%. Cystatin C is stable for 3 days in room temperature and 14 days in +4C. The assay did not show any major interference with haemoglobin. The urinary cystatin C showed good precision and performance characteristics by measurements using PETIA all of which is a necessary qualification for a biomarker at a 24-h running routine laboratory.

Thesis (Ph. D.)--University of Missouri--Columbia, 2001. / "May 2001." Typescript. Vita. Includes bibliographical references (leaves 145-152). Also available on the Internet.

Asthma, current detection methods and metabolites proposed as asthma markers are described. The limitation of the disease diagnosis is outlined and metabolomics is introduced as the approach carried out within this research with the potential to measure the group metabolites that characterise the metabolic responses of a biological system to a specific disease. Chemistry underlying breathing, current breath collection and analytical techniques are described as well as detection and data processing technology associated within our research. A work-flow for the collection, analysis and processing of exhaled breath samples in respiratory diseases is described. The non-invasive sampling method allows collection of exhaled breath samples on children and adults without experiencing any discomfort. The analysis of exhaled breath samples using thermal desorption gas chromatography mass spectrometry outlines the use of retention index for the alignment of VOCs retention time shifting over time. This methodology enables the creation of a breath matrix for multivariate analysis data processing where each VOC is defined by retention index and most intense fragments of the mass spectrum. This methodology is tested in two cohorts of participants: paediatric asthma and severe asthmatic participants whose breath profiles are compared against healthy controls and within the two asthmatic phenotypes to prospect the markers that differentiate between the different groups. Eight candidate markers are identified to discriminate between asthmatic children and healthy children and seven markers between asthmatics undergoing therapy and healthy controls. The database from severe and paediatric asthma is compared, establishing seven non-age related markers between the two groups. A new interface is developed for the faster analysis of exhaled breath samples using thermal desorption ion mobility mass spectrometry. The interface front end has been modified and optimised to achieve the best sensitivity and resolution of VOCs in exhaled breath. A preliminary study carried out in a small cohort of volunteers shows the feasibility of the technique for the differentiation of asthmatic and healthy adults.

A study was conducted in 2004 and 2005 to determine the effect of liquid hog manure fertilization on greenhouse gas emissions from the surface of a grassland pasture in south-eastern Manitoba. The objectives of this research were to determine the effects of manure application, itstiming and soil moisture on greenhouse gas emissions from pasture soil, cattle dung and urine patches. Nitrous oxide (N2O), methane (CH4) and carbon dioxide (CO2) emissions were determined from grassland soil surface, and from cattle dung and artificial urine patches. Liquid hog manure treatments were no manure (Control); 153 kg ha-1 of available-nitrogen (N) (two year average) in spring (Spring); and 149 kg ha-1 as half-rate applications in fall and spring (Split). Four field experiments were conducted on grassland plots. The static-vented chamber technique was used to estimate gas emission rates. Two of the experiments focused on the effects of manure application timing and soil moisture on greenhouse gas emissions from the grassland soil surface. The other two experiments focused on the effects of manure application and soil moisture on greenhouse gas emissions from cattle dung and artificial urine patches. Fresh cattle dung was collected from steers grazing adjacent pastures receiving the same three manure treatments. Artificial cattle urine treatments were generated by converting blood urea concentrations of the steers into urine-N concentrations. Manure application increased (P≤0.01) cumulative N2O emissions from the grassland soil surface with Control, Split and Spring treatments averaging 7, 43 and 120 mg N2O-N m-2, respectively. Of the two manure treatments, the Spring treatment emitted higher (P≤0.10) N2O emissions than the Split treatment. Soil moisture was a major factor influencing the quantity and type of greenhouse gas emissions, with saturated areas emitting CH4 during warm periods, whereas drier areas emitted N2O. Nitrous oxide emissions from these dry areas were higher in manure-treated plots. Spring application increased root density by 45% in the top 5 cm of soil compared to the Control. An increase in soil organic carbon with root density may offset any increase in greenhouse gas emissions caused by manure treatment. Cattle dung from Split and Spring treatments had higher cumulative N2O emissions (30 and 82 mg N2O-N m-2, respectively) compared to dung from Control pastures (6 mg N2O-N m-2) over two study years. Dung from the Spring treatment emitted more N2O (P≤0.01) than the other two treatments. All cattle dung patches emitted CH4 after deposition though unaffected by manure treatment. Artificial urine having highest N concentration had greater (P≤0.05) cumulative N2O emissions (690 mg N2O-N m-2) than urine with the lowest N concentration (170 mg N2O-N m-2). Drier soil locations emitted more N2O from cattle dung and artificial urine patches than wetter areas. This study demonstrated that Split application of liquid hog manure to grassland emitted less N2O than a complete application in spring. Moisture greatly affected the location of N2O and CH4 emissions. Drier areas emitted more N2O than wetter ones. Particularly, the findings indicate a need to assess grassland on periodically saturated soils as sources rather than sinks for CH4. Application of manure increased greenhouse gas emissions from cattle dung and urine patches with urine potentially having the greatest impact because of their higher emissions of N2O. An increase in root growth seems to offset greenhouse gas emissions from manure application.