Salivary proteomics and biomarkers in neurology and psychiatry

Biomarkers are greatly needed in the fields of neurology and psychiatry, to provide objective and earlier diagnoses of CNS conditions. Proteomics and other omics MS-based technologies are tools currently being utilized in much recent CNS research. Saliva is an interesting alternative biomaterial for the proteomic study of CNS disorders, with several advantages. Collection is noninvasive and saliva has many proteins. It is easier to collect than blood and can be collected by professionals without formal medical training. For psychiatric and neurological patients, supplying a saliva sample is less anxiety-provoking than providing a blood sample, and is less embarrassing than producing a urine specimen. The use of saliva as a biomaterial has been researched for the diagnosis of and greater understanding of several CNS conditions, including neurodegenerative diseases, autism, and depression. Salivary biomarkers could be used to rule out nonpsychiatric conditions that are often mistaken for psychiatric/neurological conditions, such as fibromyalgia, and potentially to assess cognitive ability in individuals with compromised brain function. As MS and omics technology advances, the sensitivity and utility of assessing CNS conditions using distal human biomaterials such as saliva is becoming increasingly possible.     

Proteomics and genomics of urinary bladder cancer

Urinary bladder cancer is the fifth most common cancer in the United States. The National Cancer Institute estimates that the incidence rates will be 68 810 and the mortality rate will be 14 100 in the year 2008. Although the gold standards cytology and cystoscopy are specific for diagnosing bladder cancer, the former lacks the sensitivity to detect low‐grade tumors and the latter is very invasive and expensive. Therefore, scientists are interested in identifying reliable non‐invasive biomarkers that could be utilized in screening, leading to early detection and/or in predicting the progression of superficial tumors to invasive higher‐stage lesions with high specificity and sensitivity. Several biomarkers that indicate changes in the expression of proteins associated with increased risk have been identified. The purpose of this analysis is to provide an overview of the studies that have been conducted during the last decade that identify diagnostic and prognostic biomarkers via proteomic and genomic advancements.     

A unified sample preparation protocol for proteomic and genomic profiling of cervical swabs to identify biomarkers for cervical cancer screening

Cervical cancer screening is ideally suited for the development of biomarkers due to the ease of tissue acquisition and the well-established histological transitions. Furthermore, cell and biologic fluid obtained from cervix samples undergo specific molecular changes that can be profiled. However, the ideal manner and techniques for preparing cervical samples remains to be determined. To address this critical issue a patient screening protein and nucleic acid collection protocol was established. RNAlater was used to collect the samples followed by proteomic methods to identify proteins that were differentially expressed in normal cervical epithelial versus cervical cancer cells. Three hundred ninety spots were identified via 2-D DIGE that were expressed at either higher or lower levels (>three-fold) in cervical cancer samples. These proteomic results were compared to genes in a cDNA microarray analysis of microdissected neoplastic cervical specimens to identify overlapping patterns of expression. The most frequent pathways represented by the combined dataset were: cell cycle: G2/M DNA damage checkpoint regulation; aryl hydrocarbon receptor signaling; p53 signaling; cell cycle: G1/S checkpoint regulation; and the ER stress pathway. HNRPA2B1 was identified as a biomarker candidate with increased expression in cancer compared to normal cervix and validated by Western blot.     

Supercritical fluid extracts from brown coal lithotypes and their group components—molecular composition of non-polar compounds

The two Tertiary brown coals, i.e. xylitic brown coal (XBC) and humodetritous brown coal (HBC), from the ‘Turów’ and ‘Konin’ deposits in Poland and their group components were investigated on their behaviour to Soxhlet and supercritical fluid (SFE process) extractions. The raw coals were Soxhlet pre-extracted to remove unbound lipids (bitumens). Humic acids, lignin and cellulose were obtained from the pre-extracted XBC, while humic acids and residual coal were separated from the HBC. Raw coals and their group components were extracted under supercritical conditions with toluene (653 K, 8 MPa, 1 h). The yield of non-polar fractions from the Soxhlet and supercritical extracts were determined and analysed by gas chromatography-mass spectrometry (GC-MS).The diversity in biomarker compositions (n-alkanes, n-alkylbenzenes, iso-alkanes, fatty acids, diterpanes and pentacyclic triterpanes and their aromatised derivatives, methylphenanthrenes) of the supercritical fluid extracts (SFEs) were compared and contrasted with the Soxhlet extracts. The biomarker geochemical indices in Soxhlet extracts are typical of low maturity, while these released by SFE process have characteristic of a higher maturation level. The Soxhlet extracts of both coals show a dominance of 17α(H),21β(H)-homohopane with the precursor configuration of 22R. The pentacyclic triterpanes found in the SFEs are diverse structures, i.e. 17α(H),21β(H)-hopanes, moretanes and small amounts of benzohopanes, as well as geochemically unaltered homohopane with the biological configuration of 17β(H),21β(H)-(22R). Isopimarane is the characteristic of SFEs from humic acids of the coals. Partially aromatised pentacyclic triterpanes related to α- and β-amyrin are characteristic of free lipids and SFEs of humic acid fractions from both coals. The diversity in the phenanthro[4,5-bcd]furan, methylphenanthrenes and 2-methylanthracene compositions is dependent on chemical nature of the brown coal group component.     

Multivariate statistical analysis of diamondoid and biomarker data from Brazilian basin oil samples

Diamondoids are compounds commonly found in oils. Highly stable, they are more resistant to thermal and biological destruction than other hydrocarbons. Diamondoid-derived parameters have been used to assess the degree of thermal evolution and the extent of secondary cracking of light oils and condensates and in the identification of mixtures of oils from distinct migration pulses. Twenty-one samples of oils from five sedimentary basins of Brazilian continental margin were selected and studied on diamondoids and biomarkers. The oil samples were analyzed by gas chromatography coupled to mass spectrometry and principal components analysis (PCA) aiming to correlate biomarker and diamondoid parameters. Diamondoid and biomarker ratios analyzed by PCA led to a more accurate assessment of the maturity of the oils. The combination of diamondoid data with biomarker parameters also allowed to rank the oils from one of the five basins (JX basin) as the most mature oils, while oils from another basin (JE basin) were classified as mixtures formed from different pulses of migration.     

Tobacco Smoke Hemoglobin Adducts in Maternal and Fetal Blood

The maternal-fetal exchange of the potent tobacco related human carcinogen, 4-aminobiphenyl, was studied in women smokers during pregnancy. Maternal and fetal blood samples were classified as coming from nonsmokers (n=74), individuals smoking less than 1 pack of cigarettes per day (n=16), individuals smoking 1 pack of cigarettes per day (n=19), individuals smoking 1–2 packs of cigarettes per day (n=19), and individuals smoking greater than 2 packs of cigarettes per day (n=20). 4-Aminobiphenyl was extracted from both maternal and fetal blood samples using organic extractions and the released amine was qualitatively and quantitatively characterized by analysis of the samples by gas chromatographic and mass spectrometric analysis. Increasing levels of 4-aminobiphenyl - hemoglobin adducts were found as the smoking status of the women increased ranging from 144 ± 22.2 (<1 pack per day) to 633 ± 87.9 (>2 packs per day). A corresponding increase in the presence of fetal 4-aminobiphenyl hemoglobin adducts was also detected (74.3 ± 17.8; <1 pack/day to 319 ± 50.5; >2 packs/day).     

Urinary BTEX, MTBE and naphthalene as biomarkers to gain environmental exposure profiles of the general population

The aim of this work was to evaluate urinary benzene, toluene, ethylbenzene, m+p-xylene, o-xylene (BTEX), methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE), and naphthalene (NAP) as biomarkers of exposure to environmental pollutants. Personal air and urine samples from 108 subjects belonging to the Italian general population were compared. Urinary profiles were obtained by headspace gas chromatography-mass spectrometry. BTEX, MTBE, ETBE and NAP median airborne exposures during a 5-h sampling were 4.0, 25.3, 3.8, 9.3, 3.4, 3.4, < 0.8, and 3.4 µg/m³, respectively. Meanwhile, median urinary levels, as geometric means of three determinations were: 122, 397, 74, 127, 43, 49, < 15, and 46 ng/L, respectively. Urinary benzene and toluene concentrations were 4.6- and 1.2-fold higher in smokers than in non-smokers. For most chemicals, significant positive correlations between airborne exposure (log-transformed) and the corresponding biological marker (log-transformed) were found, with Pearson's r values for correlation, ranging from 0.228 to 0.396. Multiple linear regression analysis showed that the urinary level of these chemicals was influenced by personal airborne exposure, urinary creatinine, and urinary cotinine, with R² 0.733 for benzene. Urinary chemicals are useful biomarkers of environmental exposure. Given the ease of rapidly obtaining urine samples, they represent a non-invasive alternative to blood chemical analysis. The possibility of obtaining urinary exposure profiles makes this method an appealing tool for environmental epidemiology.     

Review of existing experimental methods for assessing the outcome of plant food supplementation on immune function

Plant food supplements (PFS) have become increasingly popular with respect to their consumption for improving human immune function. Despite this popularity, critical review is lacking regarding the analytical methods used to assess PFS outcome. The suitability of such methods for clinical-based studies remains particularly unclear. We undertook a literature-based review of the methods used to assess PFS outcome in immune function, to identify and assess the relevance of different technologies. Most methods described in this review adequately measured the functions of innate and adaptive immunity, were applicable to both healthy and diseased subjects, and were appropriate for assessing the benefit claims of PFS on immune function. However, the design and reporting quality of studies varied widely across trials, in some cases potentially impacting negatively on the outcomes and interpretations. Several strategies to enhance study robustness and quality were outlined, to improve the validity of the data generated in the field.     

NMR代谢组学技术在环境污染评价中的应用

随着环境污染日益严重,对环境污染评价的研究亦愈发深入。基于核磁共振(Nuclear Magnetic Resonance,NMR)代谢组学是近年发展起来的一种新型环境污染评价方法,它是以核磁共振为技术手段,从生物角度研究生物体暴露在环境污染物下,生物体内源性代谢产物谱的变化,以期预警环境污染的程度。本文将综述基于核磁共振的代谢组学在环境污染评价上的原理、研究思路、以及国内外研究新进展。