Detecting simulated patterns of lung cancer biomarkers by random network of single-walled carbon nanotubes coated with nonpolymeric organic materials

An array of chemiresistive random network of single-walled carbon nanotubes coated with nonpolymeric organic materials shows a high potential for diagnosis of lung cancer via breath samples. The sensors array shows excellent discrimination between the volatile organic compounds (VOCs) found in the breath of patients with lung cancer, relative to healthy controls, especially if the sensors array is preceded with either water extractor and/or preconcentrator of VOCs. The pattern compositions of the healthy and cancerous states were determined by gas-chromatography linked with mass-spectroscopy (GC-MS) analysis of real exhaled breath.     

Determination of phytic acid in urine by inductively coupled plasma mass spectrometry

An ICPMS method for the determination of phytic acid in human urine based on the total phosphorus measurement of purified extracts of phytic acid is described. Pretreatment of the sample is required to avoid interference in the ICPMS detection from other phosphorus compounds accompanying phytic acid in urine such as phosphate or pyrophosphate. This treatment consists of a simple filtration of the urine sample followed by complete separation of phytic acid from the mentioned phosphorus components using an anion-exchange solid-phase extraction. Separation/recovery conditions, optimized for standards of phytic acid prepared in water and artificial urine, were successfully applied to natural urine samples, resulting in adequate accuracy and precision. Linear range (0.02-0.6 mg of phytic acid L(-)(1)) and limit of detection (5 microg L(-)(1) phytic acid) are adequate for analysis of the usual amounts of phytic acid present in urine. Phosphate, pyrophosphate, and pH of urine samples at concentrations exceeding their normal physiological ranges do not affect the determination of phytic acid. Because of the simplicity, low sample requirement, and relatively high sample throughput (10 to 6 min per sample for runs between 50 and 100 samples, respectively), the present method presents the best alternative to current methods for phytic acid determination in urine. Results also show that the method is adequate for the differentiation of levels of phytic acid excretion from patients suffering from oxalocalcic urolithiasis and healthy controls, suggesting that low phytic acid concentrations in urine lead to elevated risk of oxalocalcic urolithiasis.     

Automated texture‐based characterization of fibrosis and carcinoma using low‐dose lung CT images

Lung cancer is the most common cause of cancer deaths worldwide and account for 1.38 million deaths per year. Patients with lung cancer are often misdiagnosed as pulmonary tuberculosis (TB) leading to delay in the correct diagnosis as well as exposure to inappropriate medication. The diagnosis of TB and lung cancer can be difficult as symptoms of both diseases are similar in computed tomography (CT) images. However, treating TB leads to inflammatory fibrosis in some of the patients. There comes the need of an efficient computer aided diagnosis (CAD) of the fibrosis and carcinoma diseases. To design a fully automated CAD for characterizing fibrous and carcinoma tissues without human intervention using lung CT images. The 18 subjects in this study include seven healthy, two fibrosis and eight carcinoma, and one necrosis cases. The dataset is built by CT cuts representing healthy is 113, fibrosis is 103, necrosis is 39, and carcinoma is 185 totalling 440 images. The gray‐level spatial dependence matrix and gray level run length matrix approach are used for extracting texture‐based features. These features are given to neural network classifier and statistical classifier. These classifier performances are evaluated using receiver‐operating characteristics (ROC). The proposed method characterizes these tissues without human intervention. Sensitivity, specificity, precision, and accuracy followed by ROC curves were obtained and also studied. Thus, the proposed automated image‐based classifier could act as a precursor to histopathological analysis, thereby creating a way to class specific treatment procedures.     

Comparative investigations of ultrafine crystals in urine of healthy human and lithogenic patients

The ultrafine crystals in urine of lithogenic patients and healthy human were investigated by transmission electron microscopy (TEM), selected-area electron diffraction (SAED), X-ray diffraction (XRD) and Fourier transform infrared (FT-IR) spectroscopy. Phosphates and calcium oxalate monohydrate (COM) were present in urine of both healthy human and lithogenic patients. In urine of healthy human, the ultrafine crystals were well dispersed and more rounded, their size was smaller. However, the ultrafine crystals in lithogenic urine were highly aggregated. These results were explained by the difference of the concentration and activity of urinary inhibitors. The inhibitors in healthy urine have strong ability to inhibit the growth and aggregation of urinary crystals, making the edges and tips of these crystals more rounded.     

On-line concentration and separation of proteins by capillary electrophoresis using polymer solutions

Proteins were separated in 0.6% poly(ethylene oxide) (PEO) solutions using a capillary filled with buffers prior to analysis and were detected by laser-induced native fluorescence using a pulsed Nd:YAG laser. PEO solutions entered the capillary by electroosmotic flow (EOF) during the separation. The composition and concentration of the buffer affected the adsorption of PEO molecules on the capillary surface and, consequently caused changes in the EOF. Short separation times (< 7 min) were achieved on a sample solution of five proteins in a 0.6% PEO solution containing 5 microg/mL ethidium bromide using a capillary pre-filled with 100 mM TRIS-borate (TB) buffers (pH 10,0). We also extended this method for on-line concentration and separation of proteins. Proteins dissolved in low-conductivity media stacked in both TB buffers and in PEO solutions. The peak height was proportional to the injection volume up to 2.1 microL using an 80-cm capillary filled with 400 mM TB buffers. Using large injection volumes (2.1 microL), we achieved a limit of detection (S/N = 3) of 31 pM for carbonic anhydrase, which was a 1696-fold sensitivity enhancement compared to a conventional injection method (1 kV for 10 s). In high-conductivity media (urine matrix), stacking occurred at the boundary between the sample zone and PEO solutions. A urine sample without any pretreatment was analyzed, and after stacking, several peaks were detected. Spiking the urine sample with human serum albumin (HSA) affected the fluorescent intensity of some analytes as a result of interaction with HSA.     

In situ surface sampling of biological objects and preconcentration of their volatiles for chromatographic analysis

This report describes a rolling stir bar sampling procedure for volatile organic compounds (VOCs) present on various biological surfaces. In combination with thermal desorption/gas chromatography/mass spectrometry, this analytical technique was initially tested for quantitative profiling of human skin VOCs. It is also applicable to additional hydrophobic surfaces such as agricultural products, plant materials, and bird feathers. Use of embedded internal standards provides highly reproducible and quantitative results for a wide variety of sampled trace components. The samples of collected human skin VOCs and standards were found stable under cool storage conditions for at least 14 days, making this approach suitable for field biological and agricultural studies. Additionally, this methodology appears to have potential for forensic and toxicological investigations, as suggested through the analyses of VOC profiles of the human thumb prints recovered from a nonbiological smooth surface.     

Objective measurement of isoniazid levels: practical approach for monitoring tuberculosis drug treatment adherence

Drug treatment adherence is one of the biggest issues in tuberculosis (TB) treatment. Including DOTS (Directly Observed Treatment, Short‐course) program, many programs for TB cases have yielded marginal success and the new drug resistance varieties of TB persist. During the 1960s, researchers have projected naphthoquinone–mercuric chloride (N‐M) test as the single most way to objectively track the presence of isoniazid (INH) in urine and hence treatment adherence. INH is metabolised by cytochrome P450s and usually cleared from body within 8 h. Hence, the typical INH levels in biological fluids are within 1–5 μg/ml. The N‐M test practically works at 10–50 μg/ml and the authors developed the sodium polyacrylate (SPA) as a solid surface to improve detection limit of INH by 6–10 fold. The modification involves absorbing the reagents of N‐M test on the SPA surface and lyophilised material is directly used for diagnosis. The dry beads can detect 1–2 μg/ml of INH in saliva. The test performed with spiked as well as clinical samples and the results have good correlation. This new test has a realistic chance of tracking the TB treatment adherence remotely and in that direction a system was projected for implementation.Inspec keywords: biochemistry, diseases, drugs, patient treatment, proteins, molecular biophysicsOther keywords: objective isoniazid level measurement, tuberculosis drug treatment adherence monitoring, DOTS, directly observed treatment short‐course program, naphthoquinone‐mercuric chloride test, urine, cytochrome P450s, biological fluids, sodium polyacrylate, lyophilisation, metabolism, saliva, medical diagnosis, mycobacterium TB treatment adherence     

Speciation and identification of organoselenium metabolites in human urine using inductively coupled plasma mass spectrometry and tandem mass spectrometry

A method for speciation and identification of organoselenium metabolites found in human urine samples using high performance liquid chromatography/inductively coupled plasma mass spectrometry (HPLC/ICP-MS) and tandem mass spectrometry (MS/MS) is described. Reversed-phase chromatographic separation was used for sample fractionation with the ICP-MS functioning as an element-selective detector, and six distinct selenium-containing species were detected in a human urine sample. Fractions were then collected and analyzed using a triple quadrupole mass spectrometer with electrospray ionization and collision-induced dissociation to obtain structural information. The first two fractions were identified specifically as selenomethionine and selenocystamine, estimated to be present at approximately 11 and 40 ppb, respectively. To the best of our knowledge, this is the first time these two metabolites have been positively identified in human urine.     

Determination of 16 phthalate metabolites in urine using automated sample preparation and on-line preconcentration/high-performance liquid chromatography/tandem mass spectrometry

We developed an on-line solid-phase extraction (SPE) method, coupled with isotope dilution high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) and with automated sample preparation, to simultaneously quantify 16 phthalate metabolites in human urine. The method requires a silica-based monolithic column for the initial preconcentration of the phthalate metabolites from the urine and a silica-based conventional analytical column for the chromatographic separation of the analytes of interest. It uses small amounts of urine (100 microL), is sensitive (limits of detection range from 0.11 to 0.90 ng/mL), accurate (spiked recoveries are approximately 100%), and precise (the inter- and intraday coefficients of variation are <10%). The method is not labor intensive, and, because pretreatment of the urine samples was performed automatically using an HPLC autosampler, involves minimal sample handling, thus minimizing exposure to hazardous chemicals. The method was validated on spiked, pooled urine samples and on urine samples from 43 adults with no known exposure to phthalates. The high sensitivity and high throughput (HPLC run time, including the preconcentration step, is 27 min) of this analytical method combined with the ease of use and effective automated sample preparation procedure make it suitable for large epidemiological studies to evaluate the prevalence of human exposure to phthalates.     

Altered levels of acute phase proteins in the plasma of patients with schizophrenia

Schizophrenia is a relatively common psychiatric syndrome that affects virtually all brain functions. We investigated the plasma proteome of 22 schizophrenia male patients and 20 healthy male controls using two-dimensional gel electrophoresis and mass spectrometry. In total, we have identified 66 protein spots in human plasma and found that seven of them showed altered changes in schizophrenia patients, as compared to healthy controls, which mainly were acute phase proteins (APPs). Among these APPs, haptoglobin alpha2 chain (p < 0.001), haptoglobin beta chain (p < 0.001), alpha1-antitrypsin (p = 0.001), and complement factor B precursor (p = 0.022) showed overexpression in schizophrenia patients, whereas apolipoprotein A-I (p = 0.034) and transthyretin (p = 0.035) were found to be significantly decreased in patients. In addition, the expression of apolipoprotein A-IV (p = 0.018) was significantly up-regulated in schizophrenia patients, as compared to controls. We also found these APP genes, which were differentially expressed in this study, overlap in the schizophrenia susceptibility loci. Our findings further support the hypothesis that the inflammatory response system is linked to the pathophysiology of schizophrenia.     

N‐acetylcysteine may improve residual renal function in hemodialysis patients: A pilot study

Clinical outcomes in chronic dialysis patients are highly dependent on preservation of residual renal function (RRF). N‐acetylcysteine (NAC) may have a positive effect on renal function in the setting of nephrotoxic contrast media administration. In our recent study, we showed that NAC may improve RRF in peritoneal dialysis patients. The aim of the present study was to investigate the effect of NAC on RRF in patients treated with chronic hemodialysis. Prevalent chronic hemodialysis patients with a residual urine output of at least 100 mL/24 hours were included. The patients were administered oral NAC 1200 mg twice daily for 2 weeks. Residual renal function was assessed at baseline and at the end of treatment using a midweek interdialytic urine collection for measurement of urine output and calculation of residual renal Kt/V and glomerular filtration rate (GFR). Residual GFR was measured as the mean of urea and creatinine residual renal clearance. Each patient served as his own control. Twenty patients were prospectively enrolled in the study. Administration of NAC 1200 mg twice daily for 2 weeks resulted in significant improvement in RRF: urine volume increased from 320 ± 199 to 430 ± 232 mL/24 hours (P < 0.01), residual renal Kt/V increased from 0.19 ± 0.12 to 0.29 ± 0.14 (P < 0.01), and residual GFR increased from 1.6 ± 1.6 to 2.4 ± 2.3 mL/minute/1.73 m² (P < 0.01). N‐acetylcysteine may improve RRF in patients treated with chronic hemodialysis.     

FHDT: Fuzzy and Hyco-entropy-based Decision Tree Classifier for Tuberculosis Diagnosis from Sputum Images

Tuberculosis (TB) is one of the infectious diseases spread by the infectious agent Mycobacterium tuberculosis. Sputum smear microscopy is the primary tool used for the diagnosis of pulmonary TB, but has its limitations such as low sensitivity and large observation time. Hence, an automated technique is preferred for the diagnosis of TB. This paper develops a technique for TB diagnosis based on the bacilli count by proposing Fuzzy and Hyco-entropy-based Decision Tree (FHDT) classifier using sputum smear microscopic images. The proposed technique involves three steps: segmentation, feature extraction and classification. Initially, the input sputum smear microscopic image is subjected to a colour space transformation, for which a thresholding is applied to obtain the segmented result. Important features such as length, density, area and few histogram features are extracted for FHDT-based classification that classifies the segments into few-bacilli, non-bacilli and overlapping bacilli. An entropy function, called hyco-entropy, is designed for the optimal selection of feature. For further analysis of classification, that is, to count the number in the overlapping bacilli, the fuzzy classifier is adopted. FHDT classifier is evaluated in terms of Segmentation Accuracy (SA), Mean Squared Error (MSE) and Missing Count (MC) using microscopic images taken from ZNSM-iDB, where it can attain maximum mean SA of 0.954 and mean MC of 2.4.     

Glycoblotting-assisted O-glycomics: ammonium carbamate allows for highly efficient o-glycan release from glycoproteins

Glycoblotting, high throughput method for N-glycan enrichment analysis based on the specific chemical ligation between aminooxy/hydrazide-polymers/solids and reducing N-glycans released from whole serum and cellular glycoproteins, was proved to be feasible for selective enrichment analysis of O-glycans of common (mucin) glycoproteins. We established a standard protocol of glycoblotting-based O-glycomics in combination with nonenzymatic chemical treatment to release reducing O-glycans predominantly from various glycoprotein samples. It was demonstrated that the nonreductive condition employing a simple ammonium salt, ammonium carbamate, made glycoblotting-based enrichment analysis of O-glycans possible without significant loss or unfavorable side reactions. A general workflow of glycoblotting using a hydrazide bead (BlotGlyco H), on-bead chemical manipulations, and subsequent mass spectrometry allowed for rapid O-glycomics of human milk osteopontin (OPN) and urinary MUC1 glycoproteins purified from healthy donors in a quantitative manner. It was revealed that structures of O-glycans in human milk OPN were varied with habitual fucosylation and N-acetyllactosamine units. It was also suggested that purified human urinary MUC1 was modified preferentially by sialylated O-glycans (94% of total) with 7:3 ratio of core 1 to core 2 type O-glycans. Versatility of the present strategy is evident because this method was proved to be suited for the enrichment analysis of general biological and clinical samples such as human serum and urine, cultured human cancer cells, and formalin-fixed paraffin-embedded tissue sections. It is our belief that the present protocols would greatly accelerate discovery of disease-relevant O-glycans as potential biomarkers.     

Social media for health promotion: A visual analysis of “TB proof” South Africa's Facebook page

Health promotion is an educational tool that can be used to educate and create awareness of health issues through various media forms. The purpose of this study was to explore the use of social media (TB Proof South Africa's Facebook page) in creating tuberculosis (TB) awareness. A qualitative case study approach was used to collect data from TB Proof South Africa's Facebook page. An in-depth visual analysis of TB Proof South Africa's Facebook page was carried out over a five-month period (from 1 February to June 30, 2017). The analysis of TB Proof South Africa's Facebook page was conducted in order to determine the use of social media for health promotion. Such a comprehensive analysis was aimed at determining if the visuals on this page create awareness on TB as an illness. Common themes were identified including, TB medication, TB patients and healthcare workers raising awareness on TB. The findings have potential implications for health promotion efforts using social media.     

Investigation of the potential of capillary electrophoresis with off-line matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for clinical analysis: examination of a glycoprotein factor associated with cancer cachexia

The potential of capillary electrophoresis (CE) with offline matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry has been demonstrated for the examination of a glycoprotein factor associated with cancer cachexia. A comparison of CE profiles of a healthy volunteer and a cancer patient shows the presence of additional peaks in the electropherogram of the cancer patient that could be associated with cachexia. Micropreparative CE was performed with 180-micron fused silica capillary columns with tapered ends to collect CE fractions for further identification by MALDI-TOF-MS. The analysis of crude urine samples of cancer patients exhibiting cachexia, as well as CE fractions, with MALDI-TOF-MS using ferulic acid as the matrix shows a number of characteristic ions at m/z values of approximately 24 and approximately 67 kDa. The 24-kDa peak may be identified as the cachectic factor, a glycoprotein, whereas the peak at 67 kDa is identified as albumin, which is present in urine of most patients, and to which the cachectic factor is noncovalently bound. The combined use of CE and MALDI-TOF-MS was successful in detecting cachexia in all of the patients in this study, including one patient that was in an early phase of the disease.     

Analysis of metabolomic data using support vector machines

Metabolomics is an emerging field providing insight into physiological processes. It is an effective tool to investigate disease diagnosis or conduct toxicological studies by observing changes in metabolite concentrations in various biofluids. Multivariate statistical analysis is generally employed with nuclear magnetic resonance (NMR) or mass spectrometry (MS) data to determine differences between groups (for instance diseased vs healthy). Characteristic predictive models may be built based on a set of training data, and these models are subsequently used to predict whether new test data falls under a specific class. In this study, metabolomic data is obtained by doing a (1)H NMR spectroscopy on urine samples obtained from healthy subjects (male and female) and patients suffering from Streptococcus pneumoniae. We compare the performance of traditional PLS-DA multivariate analysis to support vector machines (SVMs), a technique widely used in genome studies on two case studies: (1) a case where nearly complete distinction may be seen (healthy versus pneumonia) and (2) a case where distinction is more ambiguous (male versus female). We show that SVMs are superior to PLS-DA in both cases in terms of predictive accuracy with the least number of features. With fewer number of features, SVMs are able to give better predictive model when compared to that of PLS-DA.     

Concentration and preservation of very low abundance biomarkers in urine, such as human growth hormone (hGH), by Cibacron Blue F3G-A loaded hydrogel particles

Urine is a potential source of diagnostic biomarkers for detection of diseases, and is a very attractive means of non-invasive biospecimen collection. Nonetheless, proteomic measurement in urine is very challenging because diagnostic biomarkers exist in very low concentration (usually below the sensitivity of common immunoassays) and may be subject to rapid degradation. Hydrogel nanoparticles functionalized with Cibacron Blue F3G-A (CB) have been applied to address these challenges for urine biomarker measurement. We chose one of the most difficult low abundance, but medically relevant, hormones in the urine: human growth hormone (hGH). The normal range of hGH in serum is 1 to 10 ng/mL but the urine concentration is suspected to be a thousand times less, well below the detection limit (50 pg/mL) of sensitive clinical hGH immunoassays. We demonstrate that CB particles can capture, preserve and concentrate hGH in urine at physiological salt and urea concentrations, so that hGH can be measured in the linear range of a clinical immunometric assay. Recombinant and cadaveric hGH were captured from synthetic and human urine, concentrated and measured with an Immulite chemiluminescent immunoassay. Values of hGH less than 0.05 ng/mL (the Immulite detection limit) were concentrated to 2 ng/mL, with a urine volume of 1 mL. Dose response studies using 10 mL of urine demonstrated that the concentration of hGH in the particle eluate was linearly dependent on the concentration of hGH in the starting solution, and that all hGH was removed from solution. Thus if the starting urine volume is 100 mL, the detection limit will be 0.1 pg/mL. Urine from a healthy donor whose serum hGH concentration was 1.34 ng/mL was studied in order detect endogenous hGH. Starting from a volume of 33 mL, the particle eluate had an hGH concentration of 58 pg/mL, giving an estimated initial concentration of hGH in urine of 0.175 pg/mL. The nanotechnology described here appears to have the desired precision, accuracy and sensitivity to support large scale clinical studies of urine hGH levels.      

Crystallization rules of calcium oxalate crystals in lithogenic urine and in healthy urine in vitro

Crystallization of calcium oxalate (CaOxa) was comparatively studied in vitro in diluted lithogenic urine and in diluted healthy urine by means of scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FT-IR) spectroscopy. In healthy urine, the crystallization was a growth-controlled process in the early stage of crystallization and a nucleation-controlled process in the middle and late stage. However, the crystallization of CaOxa was always a growth-controlled process in the lithogenic urine. That is, the size of CaOxa particles grow gradually and at last large size of CaOxa stones formed. Comparing the CaOxa crystals grown in lithogenic urine and in healthy urine, three differentiations were observed. First, the average particle size of CaOxa crystals precipitated in lithogenic urine is larger than that in healthy urine. Second, the morphology of calcium oxalate monohydrate (COM) crystals changes from sharp hexagonal in lithogenic urine to round and blunt in healthy urine. Third, calcium oxalate dihydrate (COD) crystals were induced in healthy urine. The results in this study may provide important clues to cure urinary stones.     

Erythrocyte folate status and serum iron levels in patients undergoing hemodialysis

The present study was aimed to evaluate erythrocyte folate and the iron levels in diabetes and hypertension patients treated with/without hemodialysis. The effects of erythropoietin and iron treatment as well as vitamin supplementation on measured parameters were considered. The 67 controls consisted of healthy subjects (n = 22), hypertensive subjects (n = 22), and diabetic subjects (n = 23) without any renal disorder. According to primary renal disorders, the patients undergoing hemodialysis (n = 68) were classified into four groups as diabetic nephropathy, hypertensive nephropathy, reflux nephropathy or interstitial nephritis, and renal insufficiency depending on other causative factors. The mean value of erythrocyte folate levels of all patients undergoing hemodialysis was higher than the healthy control group (P < 0.05). Erythrocyte folate levels in hypertensive and diabetic nephropathy patients were higher than their own hypertensive or diabetic controls and also healthy controls (both, P < 0.05). Serum iron levels of all subgroups in hemodialysis patients were found to be similar with healthy controls (all, P > 0.05). The only significance observed within the subgroups was between diabetic controls and diabetic nephropathy patients (P < 0.05). None of the treatment or supplementation of erythropoietin, iron and vitamin affected erythrocyte folate levels (all, P > 0.05). The increase in erythrocyte folate status of patients with end stage renal diseases might be the result of sum or individual effects of causative factors such as renal pathology, compensation mechanism against renal anemia, or routine folate supplementation.