Biological Body Fluids Research Articles

Simultaneous metabolomics and lipidomics analysis based on 4in1 online analysis system reveal metabolic signatures in atherosclerotic mice

Developing efficient and comprehensive analysis methods for metabolomics and lipidomics in the biological tissues and body fluids is essential for understanding the disease mechanisms. Although various two-dimensional liquid chromatography-mass spectrometry (2D-LC-MS) methods have been proposed to expand metabolite coverage, achieving higher efficiency in integrated metabolomics and lipidomics studies remains a technical challenge. In this work, a novel 4in1 online analysis system with excellent reproducibility and mass accuracy was constructed for metabolomics and lipidomics study in various biological samples from atherosclerotic mice. This system enabled the simultaneous detection in both positive and negative ion modes with extensive polarity separation in a single analytical run. Using the 4in1 online analysis system, we identified distinct but complementary metabolic signatures associated with atherosclerosis in different biological samples. Specifically, a total of 230 and 170 differential metabolites or lipids were detected in mice plasma samples and aortic tissue samples, respectively, including glycerophospholipids, sphingolipids, fatty acyls, glycerolipids, carboxylic acids, and pyrimidine nucleosides. Additionally, atherosclerosis-related metabolic pathways involved in biosynthesis of unsaturated fatty acids, sphingolipid metabolism, cholesterol metabolism, glycerophospholipid metabolism, and choline metabolism further revealed. These findings demonstrate that the novel 4in1 online analysis system is a faithful, stable and powerful tool for comprehensive metabolomics and lipidomics studies in complex biological matrices.

The Art of Finding the Right Drug Target: Emerging Methods and Strategies.

Drug targets are specific molecules in biological tissues and body fluids that interact with drugs. Drug target discovery is a key component of drug discovery and is essential for the development of new drugs in areas such as cancer therapy and precision medicine. Traditional in vitro or in vivo target discovery methods are time-consuming and labor-intensive, limiting the pace of drug discovery. With the development of modern discovery methods, the discovery and application of various emerging technologies have greatly improved the efficiency of drug discovery, shortened the cycle time, and reduced the cost. This review provides a comprehensive overview of various emerging drug target discovery strategies, including computer-assisted approaches, drug affinity response target stability, multiomics analysis, gene editing, and nonsense-mediated mRNA degradation, and discusses the effectiveness and limitations of the various approaches, as well as their application in real cases. Through the review of the aforementioned contents, a general overview of the development of novel drug targets and disease treatment strategies will be provided, and a theoretical basis will be provided for those who are engaged in pharmaceutical science research. SIGNIFICANCE STATEMENT: Target-based drug discovery has been the main approach to drug discovery in the pharmaceutical industry for the past three decades. Traditional drug target discovery methods based on in vivo or in vitro validation are time-consuming and costly, greatly limiting the development of new drugs. Therefore, the development and selection of new methods in the drug target discovery process is crucial.

Mathematical modeling of ethanol distribution in body fluids

The aim of the study is to build a kinetic model of ethanol distribution in biological body fluids, as well as its implementation in the format of computer programs. Analytical mathematical modeling of ethanol kinetics in body fluids after single and multiple oral alcohol intake was performed. An equation for the distribution of ethanol in internal media and other body fluids has been developed based on a kinetic model of zero-order elimination with absorption and irrevocable absorption deficiency, as well as achieving an equilibrium state with first-order blood plasma. On the basis of this kinetic model, computational algorithms are constructed for estimating the concentration of ethanol in body fluids at a given time, the peak concentration of ethanol and the time of its achievement, the time of complete removal of ethanol, the beginning and end of the false elimination period. Solutions to the problem of determining the amount of ethanol taken by its concentration in the biological fluid and the level of etholemia at this point in time with the possibility of calculating their maximum absolute errors are obtained. The constructed kinetic model is generalized to any number of ethanol receptions separated by time intervals of any duration. The developed computational algorithms are implemented in the format of the computer programs 'Alcohol Calculator V 7.0" and "Alcohol Calculator V 6.0", which are recommended for use in the forensic medical expert assessment of acute alcohol intoxication.

Synthesis of poly (L-cysteine)/g-C3N4 modified glassy carbon electrodes for electrochemical detection of methotrexate as a medicine for treatment of breast cancer in pharmaceutical fluid samples

Analyzing the levels of anticancer medications in biological samples and body fluids reveals important details on the course and effects of chemotherapy. p (L-Cys)/graphitic-carbon nitride (g-C3N4)/GCE, a modified glassy carbon electrode, was created for the current study's electrochemical detection of methotrexate (MTX), a drug used to treat breast cancer, in pharmaceutical fluid samples. l-Cysteine was electro-polymerized on the surface of the g-C3N4/GCE after the g-C3N4 was first modified to prepare the p (L-Cys)/g-C3N4/GCE. Analyses of morphology and structure showed that well-crystalline p (L-Cys) on g-C3N4/GCE was successfully electropolymerized. Studying the electrochemical characteristics of p (L-Cys)/g-C3N4/GCE using CV and DPV techniques revealed a synergistic impact between g-C3N4 and l-cysteine that improved the stability and selectivity of the electrochemical oxidation of MTX while enhancing the electrochemical signal. Results showed that 7.5–780 μM was the linear range, and that 0.11841 μA/μM and 6 nM, respectively, were the sensitivity and limit of detection. The applicability of the suggested sensors was assessed using real pharmaceutical preparations, and the results showed that p (L-Cys)/g-C3N4/GCE had a high degree of precision. Five breast cancer patients who volunteered and provided prepared blood serum samples between the ages of 35 and 50 were used to examine the validity and accuracy of the proposed sensor in the current work for the determination of MTX. The results showed good recovery values (greater than 97.20%), appropriate accuracy (RSD less than 5.11%), and good agreement between the ELISA and DPV analysis results. These findings showed that p (L-Cys)/g-C3N4/GCE can be applied as a trustworthy MTX sensor for MTX level monitoring in blood samples and pharmaceutical samples.

Application value of circulating LncRNA in diagnosis, treatment, and prognosis of breast cancer.

Breast cancer is the malignant tumor with the highest incidence in women worldwide. It is highly heterogeneous, has a high incidence of drug resistance, recurrence, and metastasis, and is one of the malignant tumors with the highest mortality rate. The early diagnosis, treatment monitoring, and prognosis assessment of breast cancer are the key factors affecting the survival of patients. However, due to the lack of specific biomarkers, breast cancer is still an essential factor affecting women's quality of life and physical and mental health. Long non-coding RNA can regulate various genes and different signaling pathways and plays an essential role in the occurrence and development of tumors. Recent studies have found that the abnormal expression of circulating long non-coding RNA in serum, saliva, and other biological body fluids plays a significant role in early diagnosis, pathological classification, stage, therapeutic effect monitoring, and prognosis evaluation of breast cancer. This article will review the potential application value of circulating lncRNA in breast cancer.

Combined Use of MALDI-TOF Mass Spectrometry and 31P NMR Spectroscopy for the Analysis of (Phospho)Lipids.

Lipids are important and abundant constituents of all biological tissues and body fluids. In particular, phospholipids (PLs) constitute a major part of the cellular membrane and play a role in signal transduction, and some selected PLs are increasingly considered as potential disease markers. Unfortunately, methods of lipid analysis are less established in comparison to techniques of protein analysis. Mass spectrometry (MS) is an increasingly used technique to analyze lipids, especially in combination with electrospray ionization MS, which is the most commonly used ionization technique in lipidomics. Matrix-assisted laser desorption/ionization coupled to time-of-flight MS (MALDI-TOF MS) has itself proven to represent a useful tool in the field of lipid analysis. 31P nuclear magnetic resonance (NMR) spectroscopy, another powerful method for PL analysis, represents a direct quantitative method and does not suffer from suppression effects.This paper gives an overview of methodological aspects of MALDI-TOF MS and 31P NMR in lipid research and summarizes the specific advantages and drawbacks of both methods. In particular, suppression effects in MS will be highlighted, and possible ways to overcome this problem, e.g., the use of different matrices and separation of the relevant lipid mixture prior to analysis, will be discussed.

Quality by Design Approach for a Multicomponent Quantification Using HPLC-PDA and HPLC-MS: Application to Dosage Form and Biological Body Fluids

A multicomponent pharmaceutical that contains loratadine, paracetamol, and pseudoephedrine was quantified using HPLC-PDA. The three analytes were well-separated and quantified in the dosage form on a C-18 column using a gradient mobile phase. A quality by design strategy was followed to achieve the challenging separation. Screening and optimization steps were carried out to investigate the effect of many factors on the studied responses with a minimum number of runs. The ANOVA of the factorial model showed that % acetonitrile (factor A), flow rate (factor B), and pH (factor C) were significant. The detection of the analytes’ peaks was carried out using a PDA detector at 248nm for loratadine and paracetamol, and 214 nm for pseudoephedrine. The second method was SPE-HPLC-MS, where the three analytes and desloratadine, the active metabolite of loratadine, were quantified in spiked plasma and urine, using betamethasone valerate as an internal standard. The recovery of the analytes from body fluids was above 96%, and the LOQ was below 0.5 ng/mL. The validation of the developed HPLC-PDA method was achieved as per ICH guidelines, whereas the HPLC-MS method was validated according to FDA guidelines for bioanalytical method validation. The results were compared with the reported method, and no significant differences were found.

Kinetics of human infection with Kyasanur Forest Disease Virus

Background: Kyasanur Forest disease (KFD) was first reported in 1957 and became an emerging tick-borne viral disease of public health importance in India. However, very little is known about the host-virus interaction and pathogenesis of KFD in humans. This study described the presence, duration, and kinetics of KFDV RNA in body fluids in infected human cases. Methodology: We enrolled 76 laboratory-confirmed KFD individuals and followed them up in the study. We obtained serial samples of blood, throat swabs in viral transport medium (VTM), urine, stool, and semen during the acute and convalescent phase of KFD illness. In addition, specimens were inactivated, and nucleic acid was extracted and tested for KFDV real-time reverse transcriptase -PCR. Clinical data was also obtained from the subjects. Result: The study provides evidence of KFD virus RNA in different biological body fluids of humans. The percentage positivity of KFDV RNA in blood was 100% during the first four days of illness. PCR became negative in most cases by 7–8 days; a subset of cases (14%) had prolonged viremia for up to 15 days post-onset of illness. Relatively low copies of KFDV RNA were also detected in throat swabs and urine in the first week of illness. In addition, we detected KFDV RNA in stool samples of cases of those who had diarrhea at an early stage of infection. Conclusion: The study provides evidence of KFDV RNA in different biological body fluids, which will help understand the pathogenesis, transmission pattern and develop diagnostic algorithms of KFDV in humans. In Kyasanur Forest disease infection, the blood has more RNA copies/ml than other body fluids, and viremia may last up to two weeks post-infection.

Exploring the Arginine Methylome by Nuclear Magnetic Resonance Spectroscopy.

Protein-bound arginine is commonly methylated in many proteins and regulates their function by altering the physicochemical properties, their interaction with other molecules, including other proteins or nucleic acids. This work presents an easily implementable protocol for quantifying arginine and its derivatives, including asymmetric and symmetric dimethylarginine (ADMA and SDMA, respectively) and monomethyl arginine (MMA). Following protein isolation from biological body fluids, tissues, or cell lysates, a simple method for homogenization, precipitation of proteins, and protein hydrolysis is described. Since the hydrolysates contain many other components, such as other amino acids, lipids, and nucleic acids, a purification step using solid-phase extraction (SPE) is essential. SPE can either be performed manually using centrifuges or a pipetting robot. The sensitivity for ADMA using the current protocol is about 100 nmol/L. The upper limit of detection for arginine is 3 mmol/L due to SPE saturation. In summary, this protocol describes a robust method, which spans from biological sample preparation to NMR-based detection, providing valuable hints and pitfalls for successful work when studying the arginine methylome.

Exploring the Arginine Methylome by Nuclear Magnetic Resonance Spectroscopy

Protein-bound arginine is commonly methylated in many proteins and regulates their function by altering the physicochemical properties, their interaction with other molecules, including other proteins or nucleic acids. This work presents an easily implementable protocol for quantifying arginine and its derivatives, including asymmetric and symmetric dimethylarginine (ADMA and SDMA, respectively) and monomethyl arginine (MMA). Following protein isolation from biological body fluids, tissues, or cell lysates, a simple method for homogenization, precipitation of proteins, and protein hydrolysis is described. Since the hydrolysates contain many other components, such as other amino acids, lipids, and nucleic acids, a purification step using solid-phase extraction (SPE) is essential. SPE can either be performed manually using centrifuges or a pipetting robot. The sensitivity for ADMA using the current protocol is about 100 nmol/L. The upper limit of detection for arginine is 3 mmol/L due to SPE saturation. In summary, this protocol describes a robust method, which spans from biological sample preparation to NMR-based detection, providing valuable hints and pitfalls for successful work when studying the arginine methylome.

Diagnostic value of cortisol identification in biological body fluid in case of infectious diseases (literature review)

Disorders of control mechanisms caused by glucocorticoid hormones of adrenal cortex have a significant role in the pathogenesis of infectious diseases, first of all, due to cortisol, one of the key hormones with anti-inflammatory activity. Currently the conception about the mechanisms of cortisol influence, its functional abilities, connection with immune and nerve cells, involvement in cytokine regulation, features of free-radical oxidation has been extended. There has been identified the dependence of cortisol influence upon the isoform, amount and affinity of its receptors on target cells. The present review describes the study results concerning cortisol level in case of the most often occurring infectious diseases in children acute respiratory and intestinal infections, infectious diseases of the central nervous system. There has been noticed a considerable data variability about cortisol level in normal state and in pathological one, however, the majority of articles have detected its connection with clinical manifestations and outcomes of the diseases. The study of cortisol level in cerebrospinal fluid is of a special interest in case of neuroinfections, specifying its direct connection with the disease severity and aetiology that gives new possibilities to develop effective diagnostic criteria. In general, the literature data specifies the advanced study of disorders of hypothalamus-hypophysial-adrenal gland functioning, receptor apparatus of target cells, as well as interrelations of cortisol with immune system in case of infectious diseases to reveal new criteria for diagnostics, course prediction and disease outcome, therapy correction.

Novel enzymatic graphene oxide based biosensor for the detection of glutathione in biological body fluids

In this work, we report a novel enzymatic biosensor based on glutathione peroxidase (GSH-Px), graphene oxide (GO) and nafion for the electrochemical sensing of glutathione (GSH) in body fluids. GSH-Px was immobilized covalently via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) onto modified glassy carbon electrode (GCE) decorated with GO and nafion and successfully used for sensing of GSH in the presence of H2O2 as catalyst with Michaelis-Menten constant about 0.131 mmol/L. The active surface are of GCE improve from 0.183 cm2 to 0.225 cm2 after modification with GO. The introduced biosensor (GSH-Px/GO/nafion/GCE) was used for monitoring of GSH over the range 0.003–370.0 μM, with a detection limit of 1.5 nM using differential pulse voltammetric (DPV) method. The GSH-Px/GO/nafion/GCE was successfully applied to the determination of GSH in real samples.

Synthesis of methoxy poly(ethylene glycol)-poly(ε-caprolactone) diblock copolymers hybridized with DDAB cationic lipid as the efficient nanocarriers for in vitro delivery of lycopene into MCF-7 breast cancer cells

Given the high rate of cancer mortality in the world, trying to find appropriate therapeutic strategies is a very crucial issue. Since insulin-like growth factor −1(IGF-1) and its receptor (IGF-1R) play a significant role in tumor formation and progression, targeting them can have a vital effect on cancer treatment. Lycopene is a natural antioxidant that exerts its anti-tumor effect by regulating IGF-I signaling and cell cycle arrest, making it a suitable therapeutic agent for reducing tumor induction. However, the utilization of lycopene is limited due to its high lipophilicity and very low solubility in biological body fluids. To overcome this problem, polymer lipid hybrid nanocarriers that have a core-shell structure have received much attention in recent years. In this study, to produce lipid-hybrid nanoparticles (NPs), methoxypolyethylene glycol-polycaprolactone (mPEG-PCL) copolymers were applied for lycopene loading due to biocompatibility, biodegradability, non-toxic nature, small size, amphiphilic properties, and stability of these NPs in the circulatory system. Furthermore, the use of the cationic lipid of Dimethyldioctadecylammonium bromide (DDAB) increased the cellular uptake of nanocarriers. This study proposed mPEG-PCL-DDAB as a potent nanocarrier for drug delivery to MCF-7 breast cancer cell lines, in vitro, and their applicability for the IGF-1R silencing, as a promising anti-cancer therapeutic approach, for the first time.

“Ion Pool” Structural Ion Storage Device: A New Strategy to Collect Ions by Nanoconfinement Effects

Ion storage structure widely exists in organisms, which is used to harvesting energy in environment and converting it into ion concentration gradient to maintain complex life activities. The construction of ion storage structures relies on isolating the biological body fluids by biofilm systems, which can also be regarded as local ions confinement. Mimicking this ions storage process, an "ion pool" structural ion storage device is proposed in this research by artificial ion nanochannels, which can transform the electric power into ion concentration gradient. It is consisted of micrometer-sized ions reservoir and nanosized ions filters. Ions can be isolated within the "pool" and performed ultrahigh ions enrichment or depletion behavior deviated from bulk. Through numerical simulation by Poisson-Nernst-Planck equations, "ion pool" structural device achieves nearly 20000 rectification ratio with low surface charge. An "ion pool" structural ions storage device is also constructed with block copolymer and polyethylene terephthalate composite membranes, a super high rectification ratio of 3184.0 is achieved from the experiment, which is the highest reported so far. The ion storage efficiency of the device reaches 14.90%, which is an order of magnitude better than non-"ion pool" structural nanofluid devices.

ЗНАЧЕНИЕ СОДЕРЖАНИЯ ВЕЩЕСТВ НИЗКОЙ И СРЕДНЕЙ МОЛЕКУЛЯРНОЙ МАССЫ И ОЛИГОПЕПТИДОВ В ПАТОГЕНЕЗЕ ВЕТРЯНОЙ ОСПЫ

Endogenous intoxication syndrome is an important link in the pathogenesis of many infectious diseases. The role of low and medium molecular weight substances and oligopeptides acting as indicators of endogenous intoxication syndrome in the pathogenesis of varicella was studied, depending on the period of the disease, the severity of the course and the presence of complications. Based on this goal, 125 patients with varicella aged 18 to 49 years were examined. Determination of the content of substances of low and medium molecular weight in biological body fluids (blood plasma, red blood cells and urine) was carried out by the method of M. Ya.Malakhova (1995) on a spectrophotometer SF-46. The Lowru (1951) method was used to determine the level of oligopeptides in body fluids. The maximum level of substances of low and medium molecular weight was observed in patients with a complicated course of varicella during the height of the disease, as well as in patients with a severe course of the disease during the height. When determining the oligopeptides, the highest values were also obtained at the height of the disease in the severe course, as well as in the complicated course at admission. During the examination, the dependence of the studied indicators on the period of the disease, the severity, as well as the presence of complications was revealed. Thus, the most pronounced changes in the level of low and medium molecular weight substances and oligopeptides were observed in severe, and the least – in the mild course of varicella in all the studied media. In patients with developed complications, the disease was more severe, while the changes in the studied parameters were more pronounced.

Clinical Literature Evaluation Supports Correlation Between Oral Health And Systemic Diseases

Objective A systematic review of research literature to identify specific pro-inflammatory biomarkers that are elevated and correlate with poor oral health, provide a target for rapid diagnostic testing (RDT) that can help to improve the patient's overall health. Background A wealth of clinical research literature supports the premise that a patient's overall health is related to their oral health. Poor oral hygiene can lead to gum disease, i.e., periodontal disease (PD). There are several systemic and/or chronic diseases (SIDs) that produce elevated levels of inflammatory biomarkers. The hypothesis of this research project is the presence of specific bacterial pathogens and/or elevated biomarkers in the oral cavity are related to one or more SIDs; and early detection via testing of the oral cavity can lead to early detection of such SIDs. Detection of elevated inflammatory biomarkers in body tissues and fluids (e.g., plasma) of patients with PD could lead to development of a clinical point-of-care (POC) testing and earlier preventative treatment. To-date, there has been no systematic evaluation of outcomes from these numerous studies to identify which biomarker(s) should be targeted for a POC testing. Methods Electronic databases (e.g. PubMed, Elsevier, Google scholar) were searched for research articles containing the keywords: periodontal disease, salivary diagnostics, POC testing, systemic inflammatory diseases, and various biomarkers {e.g., IL-6, IL-1, TNF-alpha (TNF-α), and C-reactive protein (CRP)}. The data was collected and analyzed to identify relationships between bacterial pathogens and specific biomarkers present in both PD and SIDs. Results: The positive association between PD and SIDs is evident from bacterial pathogens and biomarkers that are found in biological tissues and body fluids (e.g., saliva, endothelium, brain tissue, bronchoalveolar fluid, and atherosclerotic plaques, and plasma) in inflammatory-related disease states. These diseases include cardiovascular disease, diabetes, rheumatoid arthritis, Alzheimer's disease, and pulmonary disease. The most virulent pathogens present in periodontal disease and various SIDs include: Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythensis, and Treponema denticola. Biomarkers found to be elevated in both PD and SIDs include IL-6, IL-1, TNF-α, and CRP. There have been 86 articles evaluated demonstrating the positive relationships between periodontal disease and the following: systemic inflammatory diseases (37/86), bacterial pathogens (14/86), biomarkers (29/86), and salivary diagnostics (6/86). Conclusion This study provides the foundational information to support targeted clinical studies testing this hypothesis in pharmacies, health-fairs, as well as physicians or dentist offices. There is a need for a chair-side point of care (POC) or rapid diagnostic testing (RDT) methodology to be developed for healthcare providers to take the next step towards providing the patient early intervention. The end goal is identifying the best pathogen and/or biomarker for development of a POC in order to narrow the therapeutic gap for many patients with chronic diseases by early detection and preventative care.

Estimation of Alpha-Synuclein Monomer and Oligomer Levels in the Saliva of the Children With Autism Spectrum Disorder: A Possibility for an Early Diagnosis.

BackgroundIn degenerative brain diseases like Parkinson's disease (PD), alpha-synuclein (a-syn) can be in its monomeric (a-syn-mono) or toxic oligomeric (a-syn-oligo) or as a total (a-syn-total) forms in the biological body fluids including saliva. Past research has observed major a-syn plasma variations in children with autism spectrum disorder (ASD) pointing toward brain degenerative components in their pathophysiology. No prior study has shown a-syn levels in ASD patients' saliva.Objective This study estimates the levels of alpha-synuclein monomer (a-syn-mono) and alpha-synuclein oligomer (a-syn-oligo) in the saliva of ASD affected children so that saliva can be a method for detecting disorder.Materials and methodsThis cross-sectional, multi-center study was conducted in Islamic International Medical College, Autism Resource Centre (ARC), and Step-to-learn Rehabilitation center for the slow learner in Rawalpindi. The research was performed for one year from August 2018 to August 2019. Saliva samples from 80 children (40 ASD affected children, and 40 age- and sex-comparable healthy controls) were collected. Specific anti-alpha-synuclein monomers (anti-a-syn-mono) and anti-alpha-synuclein oligomers (anti-a-syn-oligo) enzyme-linked immunosorbent assay (ELISA) kits analyzed the salivary samples. Mean ± SD were reported for quantitative data. The data between the two groups were compared using an independent t-test. The p-value of ≤ 0.05 was considered statistically significant.ResultsA total of 80 children were included in the study (n=40 ASD affected, n=40 healthy controls). The age of participating children was between four and eight years. The mean alpha-synuclein monomer level in the saliva of ASD children was 92.03 ± 117.09 pg/ml (p≤0.05), and in healthy subjects was 186.78 ± 239.31 ρg/ml. The levels of alpha-synuclein oligomer in the saliva of patients with ASD children were 0.13 ± 0.05 ng/ml (p<0.001), and in the healthy subjects was 0.33 ± 0.26 ng/ml. Both alpha-synuclein monomer and alpha-synuclein oligomer levels were low in the saliva of ASD children.ConclusionChildren with ASD had low levels of alpha-synuclein monomer and oligomer than healthy children which are unique than that of levels found in other degenerative brain diseases.

Preparation of restricted access monolithic tip via unidirectional freezing and atom transfer radical polymerization for directly extracting magnolol and honokiol from rat plasma followed by liquid chromatography analysis

In the present study, a novel strategy based on unidirectional freezing and atom transfer radical polymerization combined with activator regenerated by electron transfer (ARGET-ATRP) was applied to synthesizing orderly macroporous monolithic column with restricted-access (RA) property in a 1000μL pipette tip. The RA column was composed of hydrophobic inner column (poly(styrene-co-ethylene glycol dimethacrylate) and hydrophilic outer layer (poly-hydroxyethyl methacrylate chain) which was grafted on the hydrophobic surface by means of the second ARGET-ATRP reaction. The as-prepared RA monolithic tip was connected to a 2mL syringe for directly extracting magnolol and honokiol from rat plasma just by manually pushing operation. The surface morphology and chemical composition of the column were characterized by scanning electronic microscope, infrared spectroscopy and X-ray photoelectron spectroscopy respectively. The determined results of evaluation experiments based on the optimized solid phase extraction conditions showed that the RA column possessed good protein exclusion power, extraction recovery and reusability. The constructed RA-SPE-HPLC/UV method for simultaneously analyzing magnolol and honokiol in rat plasma was validated with quality control (QC) samples at four concentration levels. Good precision (RSDs, 3.39~11.16%) and acceptable accuracy (relative recoveries, 89.52%~108.42%) were obtained for intra- and inter-day assays. The determined results of real rat plasma as well as the standard-addition samples demonstrated the developed method with good accuracy and precision. It can be extrapolated from the experimental results that this simple and cost-efficient RA-SPE method is also suitable for directly extracting other hydrophobic constituents in biological body fluid for therapeutic drug monitoring or pharmacokinetic study.

Dichotomy of exosomes in oral squamous cell carcinoma: Prey or play!

Exosomes are nano-sized particles which belong to the family of extracellular vesicles (EVs) that are produced in the endosomal compartment of those cells which mediate intercellular communication. These particles can be found abundantly in the biological body fluids such as urine, blood, saliva, cerebrospinal fluid and breast milk. These vesicles can transfer genetic materials such as the microRNAs, noncoding RNAs, DNA and lipids by means of direct or indirect cell-to-cell interaction. Consequently, there has been lot of growing interest related to cancer exosomes as biomarkers and as potential therapeutics. There are studies done which demonstrate the exosomes in relation to cancer, by targeting specific cells and also promote the tumor progression. The other part of the spectrum has stressed the importance of exosomes stability and its potential role in targeting cancer cells through drug delivery system of anticancer molecules. The dichotomy allied with exosomes and their role in oral squamous cell carcinoma biomarkers or as therapy enhancement will be highlighted.

The effect of a neuroprotective dose of isatin or deprenyl to mice on the profile of brain isatin-binding proteins

Isatin (indol-2,3-dione), an endogenous biofactor found in the brain, peripheral tissues and biological body fluids of humans and animals, exhibits a wide range of biological and pharmacological activities. They are realized via interaction with numerous isatin-binding proteins. Some of these proteins identified during proteomic profiling of the brain are involved in the development of neurodegenerative pathology. In the context of the neuroprotective effect, the effect of isatin is comparable to the effects of deprenyl (selegiline), a pharmacological agent used for treatment of Parkinson's disease. In this study, we have investigated the effect of a single dose administration of isatin (100 mg/kg) and deprenyl (10 mg/kg) to mice on the profile of the brain isatin-binding proteins. Comparative proteomic analysis of brain isatin-binding proteins of mice treated with isatin or deprenyl resulted in identification of a representative group of proteins (n=200) sensitive to the administration of these substances. The change in the profile of isatin-binding proteins may be obviously attributed to accumulation of isatin and deprenyl in the brain and their interaction with target proteins; this prevents protein binding to the affinity sorbent. Thus identified brain isatin-binding proteins of the control animals obviously represent specific targets that interact directly with isatin (and also with deprenyl) in vivo. Isatin or deprenyl administered to animals interact with these proteins and thus inhibit their binding to the affinity sorbent (immobilized isatin analogue).