Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3517

Search results for: body fluids and urine

3517 To Individualisation of Subject, Donar, by Determination of Serological Markers from Obtain Biological Fluid at Crime Scene

Authors: Arun Kumar, Ravindra Pal Verma, Harsh Sharma, Shani Kumar


For the present study samples was collected from 20 donors with unknown blood group and secretor status had been determined from saliva by using biological fluid. ABO typing on the concentrated samples was successfully performed after 1 month of storage. Urine stained clothing samples are often submitted to forensic science laboratories for ABH blood group antigen determination. The serogenetic markers of semen stains submitted can be used to determine the origin of any of these samples. ABH blood group substances have previously been identified from urine. ABH blood group substance is low in urine in comparison with other body fluids.

Keywords: ABH blood group, crime scene, serological markers, body fluids and urine

Procedia PDF Downloads 396
3516 Bisphenol-A Concentrations in Urine and Drinking Water Samples of Adults Living in Ankara

Authors: Hasan Atakan Sengul, Nergis Canturk, Bahar Erbas


Drinking water is indispensable for life. With increasing awareness of communities, the content of drinking water and tap water has been a matter of curiosity. The presence of Bisphenol-A is the top one when content curiosity is concerned. The most used chemical worldwide for production of polycarbonate plastics and epoxy resins is Bisphenol-A. People are exposed to Bisphenol-A chemical, which disrupts the endocrine system, almost every day. Each year it is manufactured an average of 5.4 billion kilograms of Bisphenol-A. Linear formula of Bisphenol-A is (CH₃)₂C(C₆H₄OH)₂, its molecular weight is 228.29 and CAS number is 80-05-7. Bisphenol-A is known to be used in the manufacturing of plastics, along with various chemicals. Bisphenol-A, an industrial chemical, is used in the raw materials of packaging mate-rials in the monomers of polycarbonate and epoxy resins. The pass through the nutrients of Bisphenol-A substance happens by packaging. This substance contaminates with nutrition and penetrates into body by consuming. International researches show that BPA is transported through body fluids, leading to hormonal disorders in animals. Experimental studies on animals report that BPA exposure also affects the gender of the newborn and its time to reach adolescence. The extent to what similar endocrine disrupting effects are on humans is a debate topic in many researches. In our country, detailed studies on BPA have not been done. However, it is observed that 'BPA-free' phrases are beginning to appear on plastic packaging such as baby products and water carboys. Accordingly, this situation increases the interest of the society about the subject; yet it causes information pollution. In our country, all national and international studies on exposure to BPA have been examined and Ankara province has been designated as testing region. To assess the effects of plastic use in daily habits of people and the plastic amounts removed out of the body, the results of the survey conducted with volunteers who live in Ankara has been analyzed with Sciex appliance by means of LC-MS/MS in the laboratory and the amount of exposure and BPA removal have been detected by comparing the results elicited before. The results have been compared with similar studies done in international arena and the relation between them has been exhibited. Consequently, there has been found no linear correlation between the amount of BPA in drinking water and the amount of BPA in urine. This has also revealed that environmental exposure and the habits of daily plastic use have also direct effects a human body. When the amount of BPA in drinking water is considered; minimum 0.028 µg/L, maximum 1.136 µg/L, mean 0.29194 µg/L and SD(standard deviation)= 0.199 have been detected. When the amount of BPA in urine is considered; minimum 0.028 µg/L, maximum 0.48 µg/L, mean 0.19181 µg/L and SD= 0.099 have been detected. In conclusion, there has been found no linear correlation between the amount of BPA in drinking water and the amount of BPA in urine (r= -0.151). The p value of the comparison between drinking water’s and urine’s BPA amounts is 0.004 which shows that there is a significant change and the amounts of BPA in urine is dependent on the amounts in drinking waters (p < 0.05). This has revealed that environmental exposure and daily plastic habits have also direct effects on the human body.

Keywords: analyze of bisphenol-A, BPA, BPA in drinking water, BPA in urine

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3515 Body Fluids Identification by Raman Spectroscopy and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Authors: Huixia Shi, Can Hu, Jun Zhu, Hongling Guo, Haiyan Li, Hongyan Du


The identification of human body fluids during forensic investigations is a critical step to determine key details, and present strong evidence to testify criminal in a case. With the popularity of DNA and improved detection technology, the potential question must be revolved that whether the suspect’s DNA derived from saliva or semen, menstrual or peripheral blood, how to identify the red substance or aged blood traces on the spot is blood; How to determine who contribute the right one in mixed stains. In recent years, molecular approaches have been developing increasingly on mRNA, miRNA, DNA methylation and microbial markers, but appear expensive, time-consuming, and destructive disadvantages. Physicochemical methods are utilized frequently such us scanning electron microscopy/energy spectroscopy and X-ray fluorescence and so on, but results only showing one or two characteristics of body fluid itself and that out of working in unknown or mixed body fluid stains. This paper focuses on using chemistry methods Raman spectroscopy and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to discriminate species of peripheral blood, menstrual blood, semen, saliva, vaginal secretions, urine or sweat. Firstly, non-destructive, confirmatory, convenient and fast Raman spectroscopy method combined with more accurate matrix-assisted laser desorption/ionization time-of-flight mass spectrometry method can totally distinguish one from other body fluids. Secondly, 11 spectral signatures and specific metabolic molecules have been obtained by analysis results after 70 samples detected. Thirdly, Raman results showed peripheral and menstrual blood, saliva and vaginal have highly similar spectroscopic features. Advanced statistical analysis of the multiple Raman spectra must be requested to classify one to another. On the other hand, it seems that the lactic acid can differentiate peripheral and menstrual blood detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, but that is not a specific metabolic molecule, more sensitivity ones will be analyzed in a forward study. These results demonstrate the great potential of the developed chemistry methods for forensic applications, although more work is needed for method validation.

Keywords: body fluids, identification, Raman spectroscopy, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

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3514 Two Years Retrospective Study of Body Fluid Cultures Obtained from Patients in the Intensive Care Unit of General Hospital of Ioannina

Authors: N. Varsamis, M. Gerasimou, P. Christodoulou, S. Mantzoukis, G. Kolliopoulou, N. Zotos


Purpose: Body fluids (pleural, peritoneal, synovial, pericardial, cerebrospinal) are an important element in the detection of microorganisms. For this reason, it is important to examine them in the Intensive Care Unit (ICU) patients. Material and Method: Body fluids are transported through sterile containers and enriched as soon as possible with Tryptic Soy Broth (TSB). After one day of incubation, the broth is poured into selective media: Blood, Mac Conkey No. 2, Chocolate, Mueller Hinton, Chapman and Saboureaud agar. The above selective media are incubated directly for 2 days. After this period, if any number of microbial colonies are detected, gram staining is performed. After that, the isolated organisms are identified by biochemical techniques in the automated Microscan system (Siemens) and followed by a sensitivity test on the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by Kirby Bauer-based plate test. Results: In 2017 the Laboratory of Microbiology received 60 samples of body fluids from the ICU. More specifically the Microbiology Department received 6 peritoneal fluid specimens, 18 pleural fluid specimens and 36 cerebrospinal fluid specimens. 36 positive cultures were tested. S. epidermidis was identified in 18 specimens, S. haemolyticus in 6, and E. faecium in 12. Conclusions: The results show low detection of microorganisms in body fluid cultures.

Keywords: body fluids, culture, intensive care unit, microorganisms

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3513 The Presence of Ochratoxin a in Breast-Milk, Urine and Serum of Lactating Women

Authors: Magdalena Twaruzek, Karolina Ropejko


Mycotoxins are secondary metabolites of molds. Ochratoxin A (OTA) is the most common in the Polish climate. It is produced by fungi of the genera Aspergillus and Penicillium. It is produced as a result of improper food storage. It is present in many products that are consumed both by humans and animals: cereals, wheat gluten, coffee, dried fruit, wine, grape juice, spices, beer, and products based on them. OTA is nephrotoxic, hepatotoxic, potentially carcinogenic, and teratogenic. OTA mainly enters an organism by oral intake. The aim of the study was to detect the presence of OTA in milk, urine, and serum of lactating women. A survey was also conducted regarding the daily diet of women. The research group consisted of 32 lactating women (11 were the donors from the Milk Bank in Toruń, the other 21 were recruited for this study). Results of the analysis showed the occurrence of OTA only in 3 milk samples (9.38%). The minimum level was 0.01 ng/ml, while the maximum 0.018 ng/ml and the mean 0.0013 ng/ml. Twenty-six urine samples (81.25%) were OTA positive, with minimum level 0.013 ng/ml, maximum level 0.117 ng/ml and mean 0.0192 ng/ml. Also, all 32 serum samples (100%) were contaminated by OTA, with a minimum level of 0.099 ng/ml, a maximum level of 2.38 ng/ml, and a mean of 0.4649 ng/ml. In the case of 3 women, OTA was present in all tested body fluids. Based on the results, the following conclusions can be drawn: the breast-milk of women in the study group is slightly contaminated with ochratoxin A. Ten samples of urine contained ochratoxin A above its average content in tested samples. Moreover, serum of 8 women contains ochratoxin A at a level above the average content of this mycotoxin in tested samples. The average ochratoxin A level in serum in the presented studies was 0.4649 ng/ml, which is much lower than the average serum ochratoxin A level established in several countries in the world, i.e., 0.7 ng/ml. Acknowledgment: This study was supported by the Polish Minister of Science and Higher Education under the program 'Regional Initiative of Excellence' in 2019 - 2022 (Grant No. 008/RID/2018/19).

Keywords: breast-milk, urine, serum, contamination, ochratoxin A

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3512 Methylation Profiling and Validation of Candidate Tissue-Specific Differentially Methylated Regions for Identification of Human Blood, Saliva, Semen and Vaginal Fluid and Its Application in Forensics

Authors: Meenu Joshi, Natalie Naidoo, Farzeen Kader


Identification of body fluids is an essential step in forensic investigation to aid in crime reconstruction. Tissue-specific differentially methylated regions (tDMRs) of the human genome can be targeted to be used as biomarkers to differentiate between body fluids. The present study was undertaken to establish the methylation status of potential tDMRs in blood, semen, saliva, and vaginal fluid by using methylation-specific PCR (MSP) and bisulfite sequencing (BS). The methylation statuses of 3 potential tDMRS in genes ZNF282, PTPRS, and HPCAL1 were analysed in 10 samples of each body fluid. With MSP analysis, the ZNF282, and PTPRS1 tDMR displayed semen-specific hypomethylation while HPCAL1 tDMR showed saliva-specific hypomethylation. With quantitative analysis by BS, the ZNF282 tDMR showed statistically significant difference in overall methylation between semen and all other body fluids as well as at individual CpG sites (p < 0.05). To evaluate the effect of environmental conditions on the stability of methylation profiles of the ZNF282 tDMR, five samples of each body fluid were subjected to five different forensic simulated conditions (dry at room temperature, wet in an exsiccator, outside on the ground, sprayed with alcohol, and sprayed with bleach) for 50 days. Vaginal fluid showed highest DNA recovery under all conditions while semen had least DNA quantity. Under outside on the ground condition, all body fluids except semen showed a decrease in methylation level; however, a significant decrease in methylation level was observed for saliva. A statistical significant difference was observed for saliva and semen (p < 0.05) for outside on the ground condition. No differences in methylation level were observed for the ZNF282 tDMR under all conditions for vaginal fluid samples. Thus, in the present study ZNF282 tDMR has been identified as a novel and stable semen-specific hypomethylation marker.

Keywords: body fluids, bisulphite sequencing, forensics, tDMRs, MSP

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3511 Growth of Struvite Crystals in Synthetic Urine Using Magnesium Nitrate

Authors: Reneiloe Seodigeng, John Kabuba, Hilary Rutto, Tumisang Seodigeng


Urine diversion toilets have become popular as a means of solving the challenges in sanitation. As a result, the source-separated urine must be adequately treated so that it can be disposed of safely and valuable struvite can be extracted for use as fertilizer. In this study, synthetic urine was prepared, and struvite crystallisation experiments carried out using magnesium nitrate. The effect of residence time on crystal growth was studied. At residence time of 10, 30 and 60 minutes, mean particle sizes were 17, 34 and 53 µm showing that with higher residence times, larger crystal sizes can be achieved. SEM analysis of the crystal showed that the resultant crystals had the typical morphology of struvite crystals.

Keywords: struvite, magnesium nitrate, crystallisation, urine treatment

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3510 Nafion Multiwalled Carbon Nano Tubes Composite Film Modified Glassy Carbon Sensor for the Voltammetric Estimation of Dianabol Steroid in Pharmaceuticals and Biological Fluids

Authors: Nouf M. Al-Ourfi, A. S. Bashammakh, M. S. El-Shahawi


The redox behavior of dianabol steroid (DS) on Nafion Multiwalled Carbon nano -tubes (MWCNT) composite film modified glassy carbon electrode (GCE) in various buffer solutions was studied using cyclic voltammetry (CV) and differential pulse- adsorptive cathodic stripping voltammetry (DP-CSV) and successfully compared with the results at non modified bare GCE. The Nafion-MWCNT composite film modified GCE exhibited the best electrochemical response among the two electrodes for the electro reduction of DS that was inferred from the EIS, CV and DP-CSV. The modified sensor showed a sensitive, stable and linear response in the concentration range of 5 – 100 nM with a detection limit of 0.08 nM. The selectivity of the proposed sensor was assessed in the presence of high concentration of major interfering species. The analytical application of the sensor for the quantification of DS in pharmaceutical formulations and biological fluids (urine) was determined and the results demonstrated acceptable recovery and RSD of 5%. Statistical treatment of the results of the proposed method revealed no significant differences in the accuracy and precision. The relative standard deviations for five measurements of 50 and 300 ng mL−1 of DS were 3.9 % and 1.0 %, respectively.

Keywords: dianabol steroid, determination, modified GCE, urine

Procedia PDF Downloads 155
3509 Diagnostic Evaluation of Urinary Angiogenin (ANG) and Clusterin (CLU) as Biomarker for Bladder Cancer

Authors: Marwa I. Shabayek, Ola A. Said, Hanan A. Attaia, Heba A. Awida


Bladder carcinoma is an important worldwide health problem. Both cystoscopy and urine cytology used in detecting bladder cancer suffer from drawbacks where cystoscopy is an invasive method and urine cytology shows low sensitivity in low grade tumors. This study validates easier and less time-consuming techniques to evaluate the value of combined use of angiogenin and clusterin in comparison and combination with voided urine cytology in the detection of bladder cancer patients. This study includes malignant (bladder cancer patients, n= 50), benign (n=20), and healthy (n=20) groups. The studied groups were subjected to cystoscopic examination, detection of bilharzial antibodies, urine cytology, and estimation of urinary angiogenin and clusterin by ELISA. The overall sensitivity and specifcity were 66% and 75% for angiogenin, 70% and 82.5% for clusterin and 46% and 80% for voided urine cytology. Combined sensitivity of angiogenin and clusterin with urine cytology increased from 82 to 88%.

Keywords: angiogenin, bladder cancer, clusterin, cytology

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3508 Studies on the Feasibility of Cow’s Urine as Non-Conventional Energy Sources

Authors: Raj Kumar Rajak, Bharat Mishra


Bio-batteries represent an entirely new long-term, reasonable, reachable, and eco-friendly approach to generation of sustainable energy. In the present experimental work, we have studied the effect of the generation of power by bio-battery using different electrode pairs. The tests show that it is possible to generate electricity using cow’s urine as an electrolyte. C-Mg electrode pair shows maximum Voltage and Short Circuit Current (SCC), while C-Zn electrode pair shows less Open Circuit Voltage (OCV) and SCC. By the studies of cow urine and different electrodes, it is found that C-Zn electrode battery is more economical. The cow urine battery with C-Zn electrode provides maximum power (707.4 mW) and durability (up to 145 h). This result shows that the bio-batteries have the potency to full fill the need of electricity demand for lower energy equipment.

Keywords: bio-batteries, cow's urine, electrodes, non-conventional

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3507 Electrochemical Behavior and Cathodic Stripping Voltammetric Determination of Dianabol Steroid in Urine at Bare Glassy Carbon Paste Electrode

Authors: N. Al-Orfi, M. S. El-Shahawi, A. S. Bashammakh


The electrochemical response of glassy carbon electrode (GCE) for the sensitive and selective determination of dianabol steroid (DS) in phosphate, Britton-Robinson (B-R) and HEPES buffers of pH 2.0 - 11, 2.0 - 11 and 6.2 - 8.0, respectively using cyclic voltammetry (CV) and differential pulse- adsorptive cathodic stripping voltammetry (DP-CSV) at bare GCE was studied. The dependence of the CV response of the developed cathodic peak potential (Ep, c), peak current (ip, c) and the current function (ip, c / υ1/2) on the scan rate (υ) at the bare GCE revealed the occurrence of electrode coupled chemical reaction of EC type mechanism. The selectivity of the proposed method was assessed in the presence of high concentrations of major interfering species e.g. uric acid, ascorbic acid, citric acid, glucose, fructose, sucrose, starch and ions Na+, K+, PO4-3, NO3- and SO42-. The recovery of the method was not significant where t(critical)=2.20 > texp=1.81-1.93 at 95% confidence. The analytical application of the sensor for the quantification of DS in biological fluids as urine was investigated. The results were demonstrated as recovery percentages in the range 95±2.5-97±4.7% with relative standard deviation (RSD) of 0.5-1.5%.

Keywords: dianabol, determination, modified electrode, urine

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3506 Perceptions of Farmers against Liquid Fertilizer Benefits of Beef Cattle Urine

Authors: Sitti Nurani Sirajuddin, Ikrar Moh. Saleh, Kasmiyati Kasim


The aim of this study was to know the perception of livestock farmers on the use of liquid organic fertilizer from urine of cattle at Sinjai Regency, South Sulawesi Province. The choice of location for a farmer group manufactures and markets liquid organic fertilizer from cattle urine. This research was conducted in May to July 2013.The population were all livestock farmers who use organic liquid fertilizer from cattle urine samples while livestock farmers who are directly involved in the manufacture of liquid organic fertilizer totaled 42 people. Data were collected through observation and interview. Data were analyzed descriptively. The results showed that the perception of livestock farmers of using liquid organic fertilizer from cattle urine provide additional revenue benefits, cost minimization farming, reducing environmental pollution which not contrary to the customs.

Keywords: liquid organic fertilizer, perceptions, farmers, beef cattle

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3505 Protective Effect of Cow Urine against Chlorpyrifos Induced-Genotoxicity and Neurotoxicity in Albino Rats

Authors: Shelly Sharma, Pooja Chadha


Humans are exposed to pesticides and insecticides either directly or indirectly. Exposure to these pesticides may lead to acute toxicity to mammals and non-target organisms. Chlorpyrifos (CPF) is a broad spectrum organophosphate pesticide widely used in various countries of the world. The aim of the present study was to assess the toxicity associated with chlorpyrifos exposure and possible mitigating effect of cow urine against genotoxic and toxic effects in rat brain induced by chlorpyrifos. For this purpose LD50 was determined and rats were orally administered with 1/8th of LD50 (19mg/kg b.wt). Brain samples were taken after 24hrs, 48hrs and 72hrs of treatment. A significant increase in the % tail DNA was observed along with the increase in MDA levels of brain tissues in chlorpyrifos treated groups as compared to control. Cow urine treated groups show decrease in DNA damage and MDA levels as compared to CPF treated group. The study indicates that cow urine has ameliorative potential against neurotoxicity and genotoxicity induced by CPF. Cow urine is considered rich in vitamin A, E and volatile fatty acids which provide antioxidant potential to it. Thus, it can be used as a genoprotective agent.

Keywords: comet assay, brain, cow urine, genotoxicity, toxicity

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3504 Simultaneous Determination of p-Phenylenediamine, N-Acetyl-p-phenylenediamine and N,N-Diacetyl-p-phenylenediamine in Human Urine by LC-MS/MS

Authors: Khaled M. Mohamed


Background: P-Phenylenediamine (PPD) is used in the manufacture of hair dyes and skin decoration. In some developing countries, suicidal, homicidal and accidental cases by PPD were recorded. In this work, a sensitive LC-MS/MS method for determination of PPD and its metabolites N-acetyl-p-phenylenediamine (MAPPD) and N,N-diacetyl-p-phenylenediamine (DAPPD) in human urine has been developed and validated. Methods: PPD, MAPPD and DAPPD were extracted from urine by methylene chloride at alkaline pH. Acetanilide was used as internal standard (IS). The analytes and IS were separated on an Eclipse XDB- C18 column (150 X 4.6 mm, 5 µm) using a mobile phase of acetonitrile-1% formic acid in gradient elution. Detection was performed by LC-MS/MS using electrospray positive ionization under multiple reaction-monitoring mode. The transition ions m/z 109 → 92, m/z 151 → 92, m/z 193 → 92, and m/z 136 → 77 were selected for the quantification of PPD, MAPPD, DAPPD, and IS, respectively. Results: Calibration curves were linear in the range 10–2000 ng/mL for all analytes. The mean recoveries for PPD, MAPPD and DAPPD were 57.62, 74.19 and 50.99%, respectively. Intra-assay and inter-assay imprecisions were within 1.58–9.52% and 5.43–9.45% respectively for PPD, MAPPD and DAPPD. Inter-assay accuracies were within -7.43 and 7.36 for all compounds. PPD, MAPPD and DAPPD were stable in urine at –20 degrees for 24 hours. Conclusions: The method was successfully applied to the analysis of PPD, MAPPD and DAPPD in urine samples collected from suicidal cases.

Keywords: p-Phenylenediamine, metabolites, urine, LC-MS/MS, validation

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3503 Applying Different Working Fluids in a Combined Power and Ejector Refrigeration Cycle with Low Temperature Heat Sources

Authors: Samad Jafarmadar, Amin Habibzadeh


A power and cooling cycle, which combines the organic Rankine cycle and the ejector refrigeration cycle supplied by waste heat energy sources, is discussed in this paper. 13 working fluids including wet, dry, and isentropic fluids are studied in order to find their performances on the combined cycle. Various operating conditions’ effects on the proposed cycle are examined by fixing power/refrigeration ratio. According to the results, dry and isentropic fluids have better performance compared with wet fluids.

Keywords: combined power and refrigeration cycle, low temperature heat sources, organic rankine cycle, working fluids

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3502 Customized Cow’s Urine Battery Using MnO2 Depolarizer

Authors: Raj Kumar Rajak, Bharat Mishra


Bio-battery represents an entirely new long term, reasonable, reachable and ecofriendly approach to production of sustainable energy. Types of batteries have been developed using MnO2 in various ways. MnO2 is suitable with physical, chemical, electrochemical, and catalytic properties, serving as an effective cathodic depolarizer and may be considered as being the life blood of the battery systems. In the present experimental work, we have studied the effect of generation of power by bio-battery using different concentrations of MnO2. The tests show that it is possible to generate electricity using cow’s urine as an electrolyte. After ascertaining the optimum concentration of MnO2, various battery parameters and performance indicates that cow urine solely produces power of 695 mW, while a combination with MnO2 (40%) enhances power of bio-battery, i.e. 1377 mW. On adding more and more MnO2 to the electrolyte, the power suppressed because inflation of internal resistance. The analysis of the data produced from experiment shows that MnO2 is quite suitable to energize the bio-battery.

Keywords: bio-batteries, cow’s urine, manganese dioxide, non-conventional

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3501 The Role of Chemokine Family, CXCL-10 Urine as a Marker Diagnosis of Active Lung Tuberculosis in HIV/AIDS Patients

Authors: Dwitya Elvira, Raveinal Masri, Rohayat Bilmahdi


Human Immunodeficiency Virus (HIV) pandemic increased significantly worldwide. The rise in cases of HIV/AIDS was also followed by an increase in the incidence of opportunistic infection, with tuberculosis being the most opportunistic infection found in HIV/AIDS and the main cause of mortality in HIV/AIDS patients. Diagnosis of tuberculosis in HIV/AIDS patients is often difficult because of the uncommon symptom in HIV/AIDS patients compared to those without the disease. Thus, diagnostic tools are required that are more effective and efficient to diagnose tuberculosis in HIV/AIDS. CXCL-10/IP-10 is a chemokine that binds to the CXCR3 receptor found in HIV/AIDS patients with a weakened immune system. Tuberculosis infection in HIV/AIDS activates chemokine IP-10 in urine, which is used as a marker for diagnosis of infection. The aim of this study was to prove whether IP-10 urine can be a biomarker diagnosis of active lung tuberculosis in HIV-AIDS patients. Design of this study is a cross sectional study involving HIV/AIDS patients with lung tuberculosis as the subject of this study. Forty-seven HIV/AIDS patients with tuberculosis based on clinical and biochemical laboratory were asked to collect urine samples and IP-10/CXCL-10 urine being measured using ELISA method with 18 healthy human urine samples as control. Forty-seven patients diagnosed as HIV/AIDS were included as a subject of this study. HIV/AIDS were more common in male than in women with the percentage in male 85.1% vs. 14.5% of women. In this study, most diagnosed patients were aged 31-40 years old, followed by those 21-30 years, and > 40 years old, with one case diagnosed at age less than 20 years of age. From the result of the urine IP-10 using ELISA method, there was significant increase of the mean value of IP-10 urine in patients with TB-HIV/AIDS co-infection compared to the healthy control with mean 61.05 pg/mL ± 78.01 pg/mL vs. mean 17.2 pg/mL. Based on this research, there was significant increase of urine IP-10/CXCL-10 in active lung tuberculosis with HIV/AIDS compared to the healthy control. From this finding, it is necessary to conduct further research into whether urine IP-10/CXCL-10 plays a significant role in TB-HIV/AIDS co-infection, which can also be used as a biomarker in the early diagnosis of TB-HIV.

Keywords: chemokine, HIV/AIDS, IP-10 urine, tuberculosis

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3500 Identification of Body Fluid at the Crime Scene by DNA Methylation Markers for Use in Forensic Science

Authors: Shirin jalili, Hadi Shirzad, Mahasti Modarresi, Samaneh Nabavi, Somayeh Khanjani


Identifying the source tissue of biological material found at crime scenes can be very informative in a number of cases. Despite their usefulness, current visual, catalytic, enzymatic, and immunologic tests for presumptive and confirmatory tissue identification are applicable only to a subset of samples, might suffer limitations such as low specificity, lack of sensitivity, and are substantially impacted by environmental insults. In addition their results are operator-dependent. Recently the possibility of discriminating body fluids using mRNA expression differences in tissues has been described but lack of long term stability of that Molecule and the need to normalize samples for each individual are limiting factors. The use of DNA should solve these issues because of its long term stability and specificity to each body fluid. Cells in the human body have a unique epigenome, which includes differences in DNA methylation in the promoter of genes. DNA methylation, which occurs at the 5′-position of the cytosine in CpG dinucleotides, has great potential for forensic identification of body fluids, because tissue-specific patterns of DNA methylation have been demonstrated, and DNA is less prone to degradation than proteins or RNA. Previous studies have reported several body fluid-specific DNA methylation markers.The presence or absence of a methyl group on the 5’ carbon of the cytosine pyridine ring in CpG dinucleotide regions called ‘CpG islands’ dictates whether the gene is expressed or silenced in the particular body fluid. Were described methylation patterns at tissue specific differentially methylated regions (tDMRs) to be stable and specific, making them excellent markers for tissue identification. The results demonstrate that methylation-based tissue identification is more than a proof-of-concept. The methodology holds promise as another viable forensic DNA analysis tool for characterization of biological materials.

Keywords: DNA methylation, forensic science, epigenome, tDMRs

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3499 Detection of Leptospira interrogans in Kidney and Urine of water Buffalo and its Relationship with Histopathological and Serological Findings

Authors: M. R. Haji Hajikolaei, A. A. Nikvand, A. R. Ghadrdan, M. Ghorbanpoor, B. Mohammadian


This study was carried out on water buffalo for detection of Leptospira interrogans in kidney and urine and its relationship with serological findings. Blood, urine and kidney samples were taken immediately after slaughter from 353 water buffalos at Ahvaz abattoir in Khouzestan province, Iran. Sera were initially screened at serum dilution of 1:100 against seven live antigens of Leptospira interrogans: pomona, hardjo, ballum, icterohemorrhagiae, tarasovi, australis and grippotyphosa using the microscopic agglutination test (MAT) and sera with positive results were titrated against reacting antigens in serial twofold dilution from 1:100 to 1:800. The samples of kidney were embedded in paraffin wax and 5µm thick sections were stained routinely with Haematoxylin and Eosin (H&E). Polymerase chain reaction (PCR) examination was done on urine and kidney by using LipL32 gene primers. Antibodies against one or more serovars at dilution >:100 were detected in sera. The most frequent reactor was hardjo (56.2%), followed by pomona (52.3%), australis (9.8%), tarassovi (5.9%), grippotyphosa (4.5%) and icterohaemorrhagiae (3.9%). The L. interrogans were detected in 43 (12.2%) of examined buffaloes, so that 26 (8.2%) of kidney tissues, 14 (4.8%) of urine samples separately and 3 (0.84%) of both kidney and urine samples were positive in PCR. From 153 (43.3%) buffaloes with positive MAT, 24 cases were positive by PCR of kidney and/or urine samples, synchronously. Renal lesions such as interstitial nephritis, acute tubular necrosis (ATN), pyelonephritis, glomerolonephritis, renal fibrosis and hydronephrosis were found in 128 (36.3%) cases. Statistical analysis indicated that there was no significant association between results of MAT, PCR and interstitial nephritis.

Keywords: leptospiral infection, PCR, MAT, histopathology, river buffalo

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3498 Heroin and Opiates Metabolites Tracing by Gas-Chromatography Isotope Ratio Mass Spectrometry

Authors: Yao-Te Yen, Chao-Hsin Cheng, Meng-Shun Huang, Shan-Zong Cyue


'Poppy-seed defense' has been a serious problem all over the world, that is because the opiates metabolites in urine are difficult to distinguish where they come from precisely. In this research, a powerful analytic method has been developed to trace the opiates metabolites in urine by Gas-Chromatography Isotope Ratio Mass Spectrometry (GC-IRMS). In order to eliminate the interference of synthesis to heroin or metabolism through human body, opiates metabolites in urine and sized heroin were hydrolyzed to morphine. Morphine is the key compound for tracing between opiates metabolites and seized heroin in this research. By matching δ13C and δ15N values through morphine, it is successful to distinguish the opiates metabolites coming from heroin or medicine. We tested seven heroin abuser’s metabolites and seized heroin in crime sites, the result showed that opiates metabolites coming from seized heroin, the variation of δ13C and δ15N for morphine are within 0.2 and 2.5‰, respectively. The variation of δ13C and δ15N for morphine are reasonable with the result of matrix match experiments. Above all, the uncertainty of 'Poppy-seed defense' can be solved easily by this analytic method, it provides the direct evidence for judge to make accurate conviction without hesitation.

Keywords: poppy-seed defense, heroin, opiates metabolites, isotope ratio mass spectrometry

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3497 Study on the Relative Factors of Introducing Table Vinegar in Reducing Urinary Tract Infection in Patients with Long-Term Indwelling Catheter

Authors: Yu-Ju Hsieh, Lin-Hung Lin, Wen-Hui Chang


This study was designed as an interventional research and intended to validate whether the introduction of drinking vinegar every day can reduce and even prevent urinary tract infection in Taiwan home stayed disabilities who using indwelling catheter. The data was collected from the subjects who have received home care case at northern Taiwan, according to the questionnaire and a medical records retroactive methodology, the subjects were informed and consent to drink 15ml of table vinegar in a daily diet, and through routine urine testing and culture study. Home care nurses would assist collecting urine at the point of before and after a meal from total 35 studied subjects per month, and total collected 4 times for testing. The results showed that when the average age of study subjects was 65.46 years and catheter indwelling time was 15 years, drinking table vinegar could inhibit the activity of E. coli O157: H7 and reduce its breeding. Before drinking table vinegar daily, the subjects’ urine pH value was 7.0-8.0, and the average was 7.5, and the urine PH value dropped to 6.5 after drinking table vinegar for a month. There were two purple urine cases whose urine were changed from purple to normal color after two weeks of drinking, and the protein and bacteria values of urine gradually improved. Urine smell unpleasant before attending to this study, and the symptom improved significantly only after 1 week, and the urine smell returned to normal ammonia and became clean after 1 month later. None of these subjects received treatment in a hospital due to urinary tract infection, and there were no signs of bleeding in all cases during this study. The subjects of this study are chronic patients with a long-term bedridden catheterization; drinking cranberry juice is an economic burden for them, and also highly prohibited for diabetes patients. By adapting to use cheaper table vinegar to acidified urine and improve its smell and ease Purple Urine Syndrome, to furthermore, proven urinary tract infection, it can also to reduce the financial burden on families, the cost of social resources and the rate of re-admission.

Keywords: table vinegar, urinary tract infection, disability patients, long-term indwelling catheter

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3496 AGEs-Aggravating Renal Lesions in C57BL/6J Mice, STZ-Induced Diabetes Nephropathy Model

Authors: Xing Lv, Hui-Qin Xu


The present study aimed to reveal the mechanism in aggravating STZ induced diabetic nephropathy (DN) by AGEs (advanced glycation end products). At the eighth day, 20 diabetic mice were randomly divided into STZ group and combination (combine AGEs with STZ) group. Simultaneously, AGEs group and normal group were set. Only mice in AGEs group, combination group were fed with high-AGEs diets. Mice diabetic conventional indicators, biochemical analysis were measured. Among the indictors, food consumptions, water intake, urine output, blood glucose, urine protein, urine creatinine, serum urea nitrogen were increased significantly in STZ, combination groups. The AGEs levels in combination group increased significantly when compared with STZ group. Weights and insulin levels in the STZ, combination groups were decreased significantly when compared with normal group, and the difference was significantly between AGEs group and STZ group. As a conclusion, AGEs play an important role in the DN development, inducing kidney damages.

Keywords: AGEs, diabetic nephropathy, serum urea nitrogen, urine protein

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3495 An Automated Magnetic Dispersive Solid-Phase Extraction Method for Detection of Cocaine in Human Urine

Authors: Feiyu Yang, Chunfang Ni, Rong Wang, Yun Zou, Wenbin Liu, Chenggong Zhang, Fenjin Sun, Chun Wang


Cocaine is the most frequently used illegal drug globally, with the global annual prevalence of cocaine used ranging from 0.3% to 0.4 % of the adult population aged 15–64 years. Growing consumption trend of abused cocaine and drug crimes are a great concern, therefore urine sample testing has become an important noninvasive sampling whereas cocaine and its metabolites (COCs) are usually present in high concentrations and relatively long detection windows. However, direct analysis of urine samples is not feasible because urine complex medium often causes low sensitivity and selectivity of the determination. On the other hand, presence of low doses of analytes in urine makes an extraction and pretreatment step important before determination. Especially, in gathered taking drug cases, the pretreatment step becomes more tedious and time-consuming. So developing a sensitive, rapid and high-throughput method for detection of COCs in human body is indispensable for law enforcement officers, treatment specialists and health officials. In this work, a new automated magnetic dispersive solid-phase extraction (MDSPE) sampling method followed by high performance liquid chromatography-mass spectrometry (HPLC-MS) was developed for quantitative enrichment of COCs from human urine, using prepared magnetic nanoparticles as absorbants. The nanoparticles were prepared by silanizing magnetic Fe3O4 nanoparticles and modifying them with divinyl benzene and vinyl pyrrolidone, which possesses the ability for specific adsorption of COCs. And this kind of magnetic particle facilitated the pretreatment steps by electromagnetically controlled extraction to achieve full automation. The proposed device significantly improved the sampling preparation efficiency with 32 samples in one batch within 40mins. Optimization of the preparation procedure for the magnetic nanoparticles was explored and the performances of magnetic nanoparticles were characterized by scanning electron microscopy, vibrating sample magnetometer and infrared spectra measurements. Several analytical experimental parameters were studied, including amount of particles, adsorption time, elution solvent, extraction and desorption kinetics, and the verification of the proposed method was accomplished. The limits of detection for the cocaine and cocaine metabolites were 0.09-1.1 ng·mL-1 with recoveries ranging from 75.1 to 105.7%. Compared to traditional sampling method, this method is time-saving and environmentally friendly. It was confirmed that the proposed automated method was a kind of highly effective way for the trace cocaine and cocaine metabolites analyses in human urine.

Keywords: automatic magnetic dispersive solid-phase extraction, cocaine detection, magnetic nanoparticles, urine sample testing

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3494 Use of a Chagas Urine Nanoparticle Test (Chunap) to Correlate with Parasitemia Levels in T. cruzi/HIV Co-Infected Patients

Authors: Yagahira E. Castro-Sesquen, Robert H. Gilman, Carolina Mejia, Daniel E. Clark, Jeong Choi, Melissa J. Reimer-Mcatee, Rocio Castro, Jorge Flores, Edward Valencia-Ayala, Faustino Torrico, Ricardo Castillo-Neyra, Lance Liotta, Caryn Bern, Alessandra Luchini


Early diagnosis of reactivation of Chagas disease in HIV patients could be lifesaving; however, in Latin American the diagnosis is performed by detection of parasitemia by microscopy which lacks sensitivity. To evaluate if levels of T. cruzi antigens in urine determined by Chunap (Chagas urine nanoparticle test) are correlated with parasitemia levels in T. cruzi/HIV co-infected patients. T. cruzi antigens in urine of HIV patients (N=55: 31 T. cruzi infected and 24 T. cruzi serology negative) were concentrated using hydrogel particles and quantified by Western Blot and a calibration curve. The percentage of Chagas positive patients determined by Chunap compared to blood microscopy, qPCR, and ELISA was 100% (6/6), 95% (18/19) and 74% (23/31), respectively. Chunap specificity was 91.7%. Linear regression analysis demonstrated a direct relationship between parasitemia levels (determined by qPCR) and urine T. cruzi antigen concentrations (p<0.001). A cut-off of > 105 pg was chosen to determine patients with reactivation of Chagas disease (6/6). Urine antigen concentration was significantly higher among patients with CD4+ lymphocyte counts below 200/mL (p=0.045). Chunap shows potential for early detection of reactivation and with appropriate adaptation can be used for monitoring Chagas disease status in T. cruzi/HIV co-infected patients.

Keywords: antigenuria, Chagas disease, Chunap, nanoparticles, parasitemia, poly N-isopropylacrylamide (NIPAm)/trypan blue particles (polyNIPAm/TB), reactivation of Chagas disease.

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3493 A Sensitive Uric Acid Electrochemical Sensing in Biofluids Based on Ni/Zn Hydroxide Nanocatalyst

Authors: Nathalia Florencia Barros Azeredo, Josué Martins Gonçalves, Pamela De Oliveira Rossini, Koiti Araki, Lucio Angnes


This work demonstrates the electroanalysis of uric acid (UA) at very low working potential (0 V vs Ag/AgCl) directly in body fluids such as saliva and sweat using electrodes modified with mixed -Ni0.75Zn0.25(OH)2 nanoparticles exhibiting stable electrocatalytic responses from alkaline down to weakly acidic media (pH 14 to 3 range). These materials were prepared for the first time and fully characterized by TEM, XRD, and spectroscopic techniques. The electrochemical properties of the modified electrodes were evaluated in a fast and simple procedure for uric acid analyses based on cyclic voltammetry and chronoamperometry, pushing down the detection and quantification limits (respectively of 2.3*10-8 and 7.6*10-8 mol L-1) with good repeatability (RSD = 3.2% for 30 successive analyses pH 14). Finally, the possibility of real application was demonstrated upon realization of unexpectedly robust and sensitive modified FTO (fluorine doped tin oxide) glass and screen-printed sensors for measurement of uric acid directly in real saliva and sweat samples, with no significant interference of usual concentrations of ascorbic acid, acetaminophen, lactate and glucose present in those body fluids (Fig. 1).

Keywords: nickel hydroxide, mixed catalyst, uric acid sensors, biofluids

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3492 The Effects of Seasonal Variation on the Microbial-N Flow to the Small Intestine and Prediction of Feed Intake in Grazing Karayaka Sheep

Authors: Mustafa Salman, Nurcan Cetinkaya, Zehra Selcuk, Bugra Genc


The objectives of the present study were to estimate the microbial-N flow to the small intestine and to predict the digestible organic matter intake (DOMI) in grazing Karayaka sheep based on urinary excretion of purine derivatives (xanthine, hypoxanthine, uric acid, and allantoin) by the use of spot urine sampling under field conditions. In the trial, 10 Karayaka sheep from 2 to 3 years of age were used. The animals were grazed in a pasture for ten months and fed with concentrate and vetch plus oat hay for the other two months (January and February) indoors. Highly significant linear and cubic relationships (P<0.001) were found among months for purine derivatives index, purine derivatives excretion, purine derivatives absorption, microbial-N and DOMI. Through urine sampling and the determination of levels of excreted urinary PD and Purine Derivatives / Creatinine ratio (PDC index), microbial-N values were estimated and they indicated that the protein nutrition of the sheep was insufficient. In conclusion, the prediction of protein nutrition of sheep under the field conditions may be possible with the use of spot urine sampling, urinary excreted PD and PDC index. The mean purine derivative levels in spot urine samples from sheep were highest in June, July and October. Protein nutrition of pastured sheep may be affected by weather changes, including rainfall. Spot urine sampling may useful in modeling the feed consumption of pasturing sheep. However, further studies are required under different field conditions with different breeds of sheep to develop spot urine sampling as a model.

Keywords: Karayaka sheep, spot sampling, urinary purine derivatives, PDC index, microbial-N, feed intake

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3491 Body Composition Analysis of Wild Labeo Bata in Relation to Body Size and Condition Factor from Chenab, Multan, Pakistan

Authors: Muhammad Naeem, Amina Zubari, Abdus Salam, Syed Ali Ayub Bukhari, Naveed Ahmad Khan


Seventy three wild Labeo bata of different body sizes, ranging from 8.20-16.00 cm total length and 7.4-86.19 g body weight, were studied for the analysis of body composition parameters (Water content, ash content, fat content, protein content) in relation to body size and condition factor. Mean percentage is found as for water 77.71 %, ash 3.42 %, fat 2.20 % and protein content 16.65 % in whole wet body weight. Highly significant positive correlations were observed between condition factor and body weight (r = 0.243). Protein contents, organic content and ash (% wet body weight) increase with increasing percent water contents for Labeo bata while these constituents (% dry body weight) and fat contents (% wet and dry body weight) have no influence on percent water. It was observed that variations in the body constituents have no association to body weight or length.

Keywords: Labeo bata, body size, body composition, condition factor

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3490 Detection of Heroin and Its Metabolites in Urine Samples: A Chemiluminescence Approach

Authors: Sonu Gandhi, Neena Capalash, Prince Sharma, C. Raman Suri


A sensitive chemiluminescence immunoassay (CIA) for heroin and its major metabolites is reported. The method is based on the competitive reaction of horseradish peroxidase (HRP)-labeled anti-MAM antibody and free drug in spiked urine samples. A hapten-protein conjugate was synthesized by using acidic derivative of monoacetyl morphine (MAM) coupled to carrier protein BSA and was used as an immunogen for the generation of anti-MAM (monoacetyl morphine) antibody. A high titer of antibody (1:64,0000) was obtained and the relative affinity constant (Kaff) of antibody was 3.1×107 l/mol. Under the optimal conditions, linear range and reactivity for heroin, mono acetyl morphine (MAM), morphine and codeine were 0.08, 0.09, 0.095 and 0.092 ng/mL respectively. The developed chemiluminescence inhibition assay could detect heroin and its metabolites in standard and urine samples up to 0.01 ng/ml.

Keywords: heroin, metabolites, chemiluminescence immunoassay, horse radish peroxidase

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3489 Simplified Modelling of Visco-Elastic Fluids for Use in Recoil Damping Systems

Authors: Prasad Pokkunuri


Visco-elastic materials combine the stress response properties of both solids and fluids and have found use in a variety of damping applications – both vibrational and acoustic. Defense and automotive applications, in particular, are subject to high impact and shock loading – for example: aircraft landing gear, firearms, and shock absorbers. Field responsive fluids – a class of smart materials – are the preferred choice of energy absorbents because of their controllability. These fluids’ stress response can be controlled by the application of a magnetic or electric field, in a closed loop. Their rheological properties – elasticity, plasticity, and viscosity – can be varied all the way from that of a liquid such as water to a hard solid. This work presents a simplified model to study the impulse response behavior of such fluids for use in recoil damping systems. The well-known Burger’s equation, in conjunction with various visco-elastic constitutive models, is used to represent fluid behavior. The Kelvin-Voigt, Upper Convected Maxwell (UCM), and Oldroyd-B constitutive models are implemented in this study. Using these models in a one-dimensional framework eliminates additional complexities due to geometry, pressure, body forces, and other source terms. Using a finite difference formulation to numerically solve the governing equation(s), the response to an initial impulse is studied. The disturbance is confined within the problem domain with no-inflow, no-outflow boundary conditions, and its decay characteristics studied. Visco-elastic fluids typically involve a time-dependent stress relaxation which gives rise to interesting behavior when subjected to an impulsive load. For particular values of viscous damping and elastic modulus, the fluid settles into a stable oscillatory state, absorbing and releasing energy without much decay. The simplified formulation enables a comprehensive study of different modes of system response, by varying relevant parameters. Using the insights gained from this study, extension to a more detailed multi-dimensional model is considered.

Keywords: Burgers Equation, Impulse Response, Recoil Damping Systems, Visco-elastic Fluids

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3488 A Steady State Characteristics of Four-Lobe Journal Bearing Lubricated with a Couple Stress Fluids in Turbulent Flow Regime

Authors: Boualem Chetti, Samir Zahaf


This paper presents the steady-state performance analysis of a four-lobe journal bearing lubricated with a couple stress fluids operating in the turbulent regime, following Constantinescu’s turbulent lubrication theory. The modified Reynolds equation is solved numerically using the finite difference method taking into consideration the effects of the turbulence and the couple stress. In this analysis, the steady-state parameters in terms of the attitude angle, load carrying capacity, side leakage and friction coefficient are determined at various values of eccentricities ratio. The computed results show that the turbulence increases the load carrying capacity, the attitude angle and the friction coefficient for a journal bearing lubricated with a Newtonian or a couple stress fluids. It is found that the turbulence has strongly influence on the steady-state performances of the four-lobe journal bearing lubricated with Newtonian fluids or a couple stress fluids.

Keywords: Four-lobe journal bearings, static characteristics, couple-stress fluids, turbulent flow

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