Human Biological Samples Research Articles

Polyphenol Consumption on Human Bile Acids Metabolism: Preliminary Data of Bile Acid Profiles in Human Biological Samples (P06-131-19)

ObjectivesBile acids (BAs) play a critical role in regulating human health through the activation of BAs receptor farnesoid X receptor (FXR) and membrane G protein coupled bile acid receptor-1 (TGR5). We aimed to develop methods to characterize BAs and their metabolites in human biological samples and characterize changes in BAs profile after chronic polyphenol consumption to help guide investigations on the potential health effects of polyphenols via BAs metabolism. MethodsPlasma, fecal and urine samples from two human studies that included berry intake were used for developing qualitative analysis of BAs using ultra high-performance liquid chromatography (UHPLC) coupled with electrospray ionization quadrupole time of flight (QTOF). The compounds were identified based on the exact mass, fragmentation pattern, available reference standards and database search. To investigate the effect of chronic polyphenol consumption on BAs composition, pooled plasma samples (fasting and postprandial 2 h, n = 6) from a chronic (45 and 90 days) strawberry supplementation study (24 g freeze dried/day) with an older population were analyzed. ResultsAmong 106 BAs and their metabolites which were tentatively identified in the samples used for method development, 70, 55, and 47 BAs species were characterized in plasma, feces and urine samples, respectively. The qualitative analysis of BAs in plasma samples from subjects following the strawberry consumption protocol detected 8 primary and 31 secondary BAs. After 90-days strawberry supplementation, two secondary BAs–glycolithocholic acid and 9(11), (5β)-cholenic acid-3α, 12α-diol were decreased to undetectable levels in the pooled fasting sample and the FRX/TGR5 agonists, including chenodeoxycholic acid, deoxycholic acid, cholic acid, glycodeoxycholic acid and taurocholic acid, showed increasing peak areas at 2 h postprandial compared to fasting. ConclusionsThe changes in BAs profiles in fasting and postprandial plasma samples after chronic strawberry feeding suggest that strawberry polyphenols may alter BAs metabolism and the FXR/TGR5 signaling. Funding SourcesThis work was funded by the California Strawberry Commission, USDA Intramural Funds and various donor funds to the Center for Nutrition Research, IIT.

A novel biostructure sorbent based on CysSB/MetSB@MWCNTs for separation of nickel and cobalt in biological samples by ultrasound assisted-dispersive ionic liquid-suspension solid phase micro extraction

An efficient method based on CysSB/MetSB@MWCNTs as a novel bio structure material was used for determination/separation of nickel and cobalt (Ni and Co) in human samples by ultrasound assisted-dispersive ionic liquid-suspension solid phase micro extraction (USA-DIL-SSPME). In this procedure, CysSB/MetSB@MWCNTs suspended in 1-butyl-2,3-dimethylimidazolium hexafluorophosphate ([BDMIM][PF6]) and mixture dispersed to 10 mL of blood samples at optimized pH by injecting. Then, the Co/Ni (II) was extracted with CysSB/MetSB@MWCNTs without any ligands and settled down in conical tube by IL [Ni/Co→:SMWCNTs]. After back extraction of ions from remaining solution, the concentration of Co/Ni was determined by electro thermal atomic absorption spectrometry (ET-AAS). By optimizing, the linear range, detection limit and enrichment factor of CysSB @MWCNTs were obtained (0.1–3.4 μg L−1; 0.08–3.2 μg L−1), (0.028 μg L−1; 0.022 μg L−1) and (50.2; 48.7) for Ni and Co ions in human biological samples, respectively (RSD<5%). The adsorption capacity of CysSB@MWCNTs for Ni and Co ions was 226.7 mg g−1 and 193.3 mg g−1, respectively which was higher than MetSB@MWCNTs. The standard reference materials (NIST, SRM) and ICP-MS were used for validation of methodology.

Collection of Human Biological Samples for the Development and Validation of New Methods of Diagnosis and for the Realization of Expertises Diagnostics in the Field of Infectious Diseases and Tropical

Collection of Human Biological Samples for the Development and Validation of New Methods of Diagnosis and for the Realization of Expertises Diagnostics in the Field of Infectious Diseases and Tropical

Rapid method for quantification of anthocyanidins and anthocyanins in human biological samples

Using simple solvent extraction and enzymatic hydrolysis, a rapid LC-MS/MS method for quantification of free and conjugated forms of anthocyanidins and anthocyanins in plasma and urine samples was developed and validated. A mixed enzymatic treatment containing β-glucuronidase (100 U mL−1) and sulfatase (2.5 U mL−1) for 5 min (37 °C; pH 6) was optimal condition for deconjugation of anthocyanidins and anthocyanins in urine and plasma samples. The LC–MS/MS allowed quantifying thirteen different anthocyanidins and anthocyanins simultaneously. The developed LC–MS/MS method was precise and accurate over multiple days and nominal concentrations. The stability assessment study confirmed that the long-term storage and/or periodic use of plasma and urine samples might have a considerable impact on the stability of some anthocyanidins. The method was successfully applied to measure anthocyanidins and anthocyanins in plasma and urine samples following consumption of acute blueberry test meals.

Review of constructive analytical methods for determining the amount of aluminum in environmental and human biological samples

Aluminum is a toxic metal and causes pollution in soil, water, and air. Afterward, a lot of patients suffer renal failure due to the accumulation of aluminum in the tissues of kidneys. Also, a high concentration of aluminum in plants tissues makes agricultural food toxic. Therefore, measuring aluminum in water, soil, air, human organs, tissues of plants and each food (or agricultural product is so necessary for protecting human health. Also, the effect of some parameters such as pH and temperature on decrease or increase in the amount of aluminum in water and other samples are stated. In this review, the analytical methods such as fluorimetric, ICP-MS, colorimetric, graphite furnace/flame atomic absorption spectrometry, etc. which have been applied for measuring the amount of aluminum (especially ) in environmental and human biological samples are assesse

Broad donor consent for human biobanks in Germany and Europe: a strategy to facilitate cross-border sharing and exchange of human biological materials and related data

Abstract Background Human biobanks are generally recognized as essential resources for effective biomedical research. All over the world biosamples and data from human subjects are collected in large biobanks. The biological material is stored long term for current and future (undetermined) research issues, which often require cross-border exchange of biosamples and related data. Content Commonly, the informed consent for research on human biospecimen is intended to cover only defined, specific research objectives. In June 2016, the biobank Task-Force of the Working Party of the German Medical Ethics Committees (WP-GMEC) updated its template for the broad use of human biological samples and related data. It complies with the current Organisation for Economic Co-operation and Development (OECD) and World Medical Association (WMA) recommendations and furnishes a framework that permits long-term storage and multi-purpose research use of human biological material and related data, including cross-border research. However, both (i) human biobanks storing and (ii) research projects requesting “broad consent” biological samples generally require an ethical approval; in addition, “broad consent” conditions should be reciprocated by making biobank processes transparent and by fostering both donor and public involvement. Outlook The broad consent template of the WP-GMEC clearly states that biological samples and data donated for medical research serve to address current and future research questions. It appears perfectly suited as a template for a Europe-wide harmonized broad consent facilitating biobank-based cross-border research.

Utility of Chimeric Mice with Humanized Liver for Predicting Human Pharmacokinetics in Drug Discovery: Comparison with in Vitro-in Vivo Extrapolation and Allometric Scaling.

Predicting human pharmacokinetics (PK) such as clearance (CL) and volume of distribution (Vd) is a critical component of drug discovery. These predictions are mainly performed by in vitro-in vivo extrapolation (IVIVE) using human biological samples, such as hepatic microsomes and hepatocytes. However, some issues with this process have arisen, such as inconsistencies between in vitro and in vivo findings; the integration of predicted CYP, non-CYP and transporter-mediated human PK; and the difficulty of evaluating very metabolically stable compounds. Various approaches to solving these issues have been reported. Allometric scaling using experimental animals has also often been used. However, this method has also shown many problems due to interspecies differences, albeit that various correction methods have been proposed. Another approach involves the production of chimeric mice with humanized liver via the transplantation of human hepatocytes into mice. The livers of these mice are repopulated mostly with human hepatocytes and express human drug-metabolizing enzymes and drug transporters, suggesting that these mice are useful for solving the issues of IVIVE and allometric scaling, and more reliably predicting human PK. In this review, we summarize human PK prediction methods using IVIVE, allometric scaling and chimeric mice with humanized liver, and discuss the utility of predicting human PK in drug discovery by comparing these chimeric mice with IVIVE and allometric scaling.

Gut microbiota derived mitochondrial inhibitors cross the blood brain barrier and localise white matter

The microbiome-gut-brain (MGB) axis is a bi-directional route of communication that exists between the brain and the microbes that reside in the gut. The MGB axis is becoming of increasing importance as significant alterations in the gut microbiota are now linked to numerous neurological conditions, however, little is currently known about the microbiome derived mediators of communication. Here we used mass spectrometry imaging (MSI), a label free imaging technique, to identify bacterial products that cross the blood brain barrier in specific pathogen free (SPF) mice. We identified two bacterial molecules abundant in white matter regions of the murine that were absent in the brain and gut in germ free (GF) mice. We have identified the primary gut microbial producers of these metabolites to be members of the Lachnospiraceae family. Both molecules were found to be structurally similar to carnitine and localise with carnitine in the SPF mouse brain. Using a primary murine cell culture model of the central nervous system white matter we show that these molecules are capable of significantly impairing mitochondrial basal respiration. Given their systemic presence in the mouse and their presence in human biological samples, these metabolites may have significant implications for diseases associated with mitochondrial dysfunction and an altered gut microbiota. These results are the first to describe a direct molecular inter-kingdom communication between prokaryotes and the mammalian brain that can facilitate functional inhibition in mammalian brain cells.

A selective probe based on 3‑aminophenyl boronic acid assembly on dithiobis(succinimidylpropionate) functionalized gold nanoparticles for sialic acid detection in human serum

Sialic acid (SA) is a notable tumor marker in human. Its increased level beyond the normal concentration in human biological samples is related to different kinds of cancers development. In this contribution, a novel colorimetric assay is reported for SA detection in form of N-acetylneuraminic acid. The sensitive detection strategy is proposed via the specific SA induced aggregation of 3‑aminophenyl boronic acid (3APBA) assembled on dithiobis(succinimidylpropionate) functionalized gold nanoparticles (AuNPs-DSP). DSP was functionalized on AuNPs through a strong AuS interaction and 3APBA was assembled on AuNPs-DSP by the amine coupling reaction between the carboxyl group of DSP on gold nanoparticles surface and the amino group of 3APBA to generate a covalent amide bond. The addition of 3APBA to buffered AuNPs-DSP generated a ruby red colloidal nanosensor. With increasing concentration of SA, a probe color turned rapidly from red to blue-gray via aggregation of AuNPs-DSP-3APBA. At optimal conditions, the absorbance ratio (A700/A520) was linear with SA concentrations ranging from 0.08 to 0.25 mM and 0.30–0.70 mM. A limit of detection of 35.0 μM was achieved which is the lowest amongst reported colorimetric methods for SA determination. The practical efficacy of the assay was demonstrated on human serum, along with no interfering effects and satisfactory accuracy and precision.

Benzoylfentanyl and parafluorobutyrfentanyl: Some analytical and metabolism data

Summary Today, New Psychoactive Substances (NPS) are increasingly being related to post-mortem cases, and this problem drastically intensifies as a result of the New Synthetic Opioids (NSOs) surge. Among NSOs, fentanyl derivatives (FDs) consist in a myriad of drugs with a high turnover. FDs detection in human biological samples is complicated by low concentrations and usual extensive metabolism. In this challenging context, this manuscript aims to report analytical data about two poorly documented FDs: benzoylfentanyl (BZF) and parafluorobutyrfentanyl (pFBF). In 2018, these FDs were identified in three samples (powders and blotting papers) in France. Nuclear magnetic resonance and liquid chromatography with high-resolution mass spectrometry detection (LC-HRMS) data including observed mass spectrometric fragmentation patterns were obtained. In addition, metabolites were investigated using human liver microsomes incubations and LC-HRMS analysis: norBZF, despropionylfentanyl and a hydroxylated-BZF were identified (and observed in hair) as BZF metabolites; norpFBF, parafluorofentanyl and a hydroxylated-pFBF were identified as pFBF metabolites. In the worrisome context of NSOs detection, these preliminary data can be useful for toxicologists.

Essential trace elemental levels (zinc, iron and copper) in the biological samples of smoker referent and pulmonary tuberculosis patients.

Tuberculosis is one of the major causes of illnesses and deaths throughout world particularly in Asia. Smoking is linked with tuberculosis recurrence and its mortality and may influence bacteriological conversion, clinical symptoms and treatment outcome. The aim of current study was to estimate association among essential trace elements {zinc (Zn), iron (Fe) and copper (Cu)} in human biological samples particularly blood, serum, scalp hair, saliva, sputum, and nasal fluid of smoking and nonsmoking pulmonary tuberculosis patients (n = 165, age ranged 16-35 years) residents of Hyderabad, Pakistan. The biological samples of age matched healthy controls were chosen as referents of both genders (n = 171) for the comparison purpose. The human biological samples were wet digested in microwave oven by 65 % HNO3 and 30 % H2O2 with (2:1) ratio. The concentrations of elements in acid digested samples were determined by atomic absorption spectrometry. The average zinc and iron concentration was lower, while level of copper was higher in the biological samples of pulmonary Tuberculosis patients as compared to referent subjects (p < .001). It was also concluded as a result of Zn and Fe deficiency combined with high contact of copper due to smoking of tobacco can be synergistic with the risk factors related with pulmonary tuberculosis.

USING THE SAMPLES OF HUMAN BIOLOGICAL MATERIALS IN THE CRIMINAL PROCEDURE: THE PRACTICE OF THE EUROPEAN COURT OF HUMAN RIGHTS

The use of modern advances in medicine to investigate crimes has caused a number of problems that require scientific reflection. In particular, today there are quite acute questions: medical intervention without the person’s consent; forced sampling of human biological materials; clinical methods, the use of which in the biological samples taking will not be regarded as violation of international standards of human rights protection; the correlation of the need for the formation of DNA profile databases and the right of the person to non-disclosure of medical information. The aim: The aim of this work is to identify and analyze the key points of the European Court of Human Rights (hereinafter referred to as the ECHR) regarding the peculiarities of retention and use of human biological material samples in the investigation of crimes, and the retention of such materials after the completion of the investigation and trial. In the preparation of the article, scientific works, the provisions of international normative acts regulating the use of human biological materials as well as the practice of the ECHR concerning the use of human biological materials in the investigation of crimes were used (8 decisions were analyzed in which the ECHR concerned the use of biological samples or related issues). In the research process to achieve the goal, a complex of general scientific and special methods of cognition was used, in particular, the comparative legal method, the system and structural method, the method of generalization, the method of analysis and synthesis, etc. The positions of the ECHR concerning the following were distinguished and generalized: a) the criteria for the permissibility of compulsory medical intervention for taking of human biological material within the framework of the crime investigation; b) the possibilities of spreading the right not to incriminate oneself on the compulsory taking of human biological materials samples; c) the retention features of cell samples and DNA information in the context of respect for the right to non-interference in the person’s private life. Obtaining and using the human material for the investigation of crimes are not a violation of the European Convention on Human Rights (hereinafter – the Convention), subject to the requirements stated in the practice of the ECHR.

Legal aspects of biobanking HBS for scientific purposes in Poland

Legal issues related to the biobanking of human biological samples are one of the extremely important areas of European law. Biobanks created in Poland as well as the Polish Biobank Network created under the auspices of the Ministry of Science and Higher Education have become a catalyst for the search for solutions and the basis of rights for the functioning of biobanks in Poland as well as the protection of donor rights. Undoubtedly, the lack of legal regulation of biobanks and biomedical research on human biological samples could become a significant problem limiting the development of biobanking and conducting scientific research in Poland. The research attempts to show how representatives of the doctrine of law, bioethics and sociologists have interpreted the principles and standards of biobank operation in Poland from basic human rights, constitutional norms and personal rights.

Electro-Catalytic Behavior of an Antiarrhythmic Drug, Procainamide and its Electro-Analytical Applications

The electrocatalytic oxidative behavior of an antiarrhythmic drug, procainamide hydrochloride (PAH) at the gold electrode surface has been examined using different voltammetric methods like cyclic, linear-sweep and differential pulse voltammetry. Voltammograms obtained in this study reveal that the electrode exhibit excellent electrocatalytic activity towards oxidation of the drug. The parameters that can affect the peak current at different pH, scan rate and concentration were evaluated. The number of electrons transferred was calculated. The current displayed a wide linear response ranging from 0.5 to 30.0 μM with a limit of detection of 56.4 nM. The impact of potential interfering agents was also studied. The electrode displayed wide advantages such as simple sample preparation, appreciable repeatability, reproducibility and also high sensitivity. Furthermore, the feasibility of the proposed method was successfully demonstrated by determining PAH in the spiked human biological sample. Keywords: Procainamide, Gold electrode, Electro-oxidation, Voltammetry, Drug determination

THE FORENSIC APPLICATION OF PROTEOMICS FOR THE STUDY OF THE TIME OF DEATH: AN OPERATIVE EXPERIMENTAL MODEL FOR PMI ESTIMATION

Post Mortem Interval (PMI) estimation is one of the most important questions the forensic pathologist must answer. To date, it is not possible to establish exactly the hour of death, but only to calculate a period (PMI), during which death most likely occurred. In the forensic field, several laboratory methods can be used to perform this calculation more accurately. However, there is still no biomarker that is universally validated and accepted by the forensic community. In the literature, researches about the application of proteomics for forensic purposes are on the increase. Proteomics is a branch of molecular biology that allows the systematic identification of the proteome from a quantitative and qualitative point of view. Below, we propose the operating model of an experimental study currently underway at the Department of Legal Medicine of the University of Catanzaro. The model is based on taking of peripheral blood samples from patients who died at the Intensive Care Unit (AO “Mater Domini” of Catanzaro). The proposed operating model has several advantages including the evaluation, for the first time, of human biological samples from the exact moment of death. The analysis would allow to identify new potential biomarkers expressed in peripheral blood and validate the forensic application of markers already known in the literature. The knowledge of the exact moment of death (time 0) would allow us to evaluate the proteomic profile more accurately on the human model, overstepping the limits of some extrinsic variables evidenced in the literature. Normal 0 14 false false false IT X-NONE X-NONE

Profiling of bisphenol A and eight of its analogues on transcriptional activity via human nuclear receptors

Several bisphenol A (BPA) analogues have been detected in environmental samples, foodstuffs, and/or human biological samples, and there is concern regarding their potential endocrine-disrupting effects. In this study, we characterized the agonistic and/or antagonistic activities of BPA and eight its analogues against human estrogen receptors (ERα/β), androgen receptor (AR), glucocorticoid receptor (GR), pregnane X receptor (PXR), and constitutive androstane receptor (CAR). All the test compounds, except for bisphenol P (BPP), showed both ERα and ERβ agonistic activities, with bisphenol AF (BPAF) being the most potent. On the other hand, BPAF and BPP showed ERα and ERβ antagonistic activities. Interestingly, their ER activities demonstrated a preference toward ERβ. All the test compounds, except for bisphenol S, showed AR antagonistic activities, with bisphenol E being the most potent. Weak GR antagonistic activities were also found in BPA and five its analogues. PXR agonistic activity was observed in the six compounds, with bisphenol Z being the most potent. Results of the CAR assay revealed that BPA and five its analogues acted as CAR inverse agonists. Taken together, these results suggested that BPA analogues demonstrate multiple effects via human nuclear receptors in a similar manner to BPA, and several analogues might have more potent endocrine-disrupting activity than does BPA.

Nucleic Acid Extraction from Human Biological Samples.

Nucleic acid isolation is often the starting point for all downstream experiments in biomedical research. It is therefore the most crucial step in any molecular technique. DNA and RNA extraction follow protocols with standardized reagents, many of which are available in quality-controlled commercial kits. Irrespective of the protocol, successful extraction of high-quality nucleic acid from biological tissues requires sufficient disruption of the tissue and cellular structures, denaturation of nucleoprotein complexes, inactivation of nucleases, and nucleic acid purification. These steps can be modified based on nucleic acid of interest and biological sample source. This chapter addresses DNA and RNA extraction from a variety of sample and tissue types, including saliva, and formalin-fixed, paraffin-embedded tissues, which are often archived in clinical pathology laboratories. Special considerations and common pitfalls of each protocol will also be discussed, as will nucleic acid quantitation techniques.

Development of Novel Dynamic Drop-to-Drop Microextraction Coupled with GC-MS for the Trace Level Screening of Phenothiazine Drug in Human Biological Samples

Development of Novel Dynamic Drop-to-Drop Microextraction Coupled with GC-MS for the Trace Level Screening of Phenothiazine Drug in Human Biological Samples

Bimetallic Fe-Cu Nanocomposites on Sand Particles for the Inactivation of Clinical Isolates and Point-of-Use Water Filtration.

Bimetallic Fe-Cu nanocomposites with an average size of 26.4 ± 4.7 nm were prepared on the surface of fine sand particles by modified coprecipitation and the chemical reduction method and were applied as an in vitro broad spectrum antimicrobial agent and recyclable hand-held water filter to sieve bacteria and metals. The size of the nanocomposites could be further reduced to 11.8 ± 1.6 nm when prepared after ball milling the sand particles, keeping the antimicrobial property intact. The results showed that the chemical nature and morphology of the nanocomposites had a great effect on both Gram-positive and Gram-negative bacteria with a minimum inhibitory concentration of 10.6 μg/mL and 13.8 μg/mL of copper, whereas the minimum bactericidal concentration was found to be 15.9 μg/mL and 21.2 μg/mL. The nanocomposites exhibited antimicrobial activity against multidrug-resistant bacteria as well as fungus isolated from different human biological samples like blood, urine, pus, and wound swabs. The nanocomposites were also capable of filtering a wide range bacteria like Acinetobacter baumannii, Escherichia coli, Salmonella typhi, Bacteroides fragilis, Salmonella paratyphi, Shigella dysenteriae, and Enterococcus faecalis, which are predominantly responsible for waterborne diseases. Further, the nanocomposites were used for the removal of hazardous metals like nickel, zinc, and lead. Leaching of copper and iron from the nanocomposites was within the permissible limit as per Bureau of Indian Standards (BIS) for Drinking Water (IS-10500-2012, second revision) as well as the International Standards for Drinking Water.

Front Cover: An accurate bacterial DNA quantification assay for HTS library preparation of human biological samples

Front Cover: An accurate bacterial DNA quantification assay for HTS library preparation of human biological samples