Identification of Mixtures of Two Types of Body Fluids Using the Multiplex Methylation System and Random Forest Models

Objective Body fluid mixtures are complex biological samples that frequently occur in crime scenes,and can provide important clues for criminal case analysis.DNA methylation assay has been applied in the identification of human body fluids,and has exhibited excellent performance in predicting single-source body fluids.The present study aims to develop a methylation SNaPshot multiplex system for body fluid identification,and accurately predict the mixture samples.In addition,the value of DNA methylation in the prediction of body fluid mixtures was further explored.Methods In the present study,420 samples of body fluid mixtures and 250 samples of single body fluids were tested using an optimized multiplex methylation system.Each kind of body fluid sample presented the specific methylation profiles of the 10 markers.Results Significant differences in methylation levels were observed between the mixtures and single body fluids.For all kinds of mixtures,the Spearman’s correlation analysis revealed a significantly strong correlation between the methylation levels and component proportions(1:20,1:10,1:5,1:1,5:1,10:1 and 20:1).Two random forest classification models were trained for the prediction of mixture types and the prediction of the mixture proportion of 2 components,based on the methylation levels of 10 markers.For the mixture prediction,Model-1 presented outstanding prediction accuracy,which reached up to 99.3%in 427 training samples,and had a remarkable accuracy of 100%in 243 independent test samples.For the mixture proportion prediction,Model-2 demonstrated an excellent accuracy of 98.8%in 252 training samples,and 98.2%in 168 independent test samples.The total prediction accuracy reached 99.3%for body fluid mixtures and 98.6%for the mixture proportions.Conclusion These results indicate the excellent capability and powerful value of the multiplex methylation system in the identification of forensic body fluid mixtures.

Critical biomarkers of hepatocellular carcinoma in body fluids and gut microbiota

Hepatocellular carcinoma(HCC)is the most prevalent primary liver cancer and one of the major causes of cancer-related death.The development of specific noninvasive or diagnostic markers from blood,urine and feces may represent a valuable tool for detecting HCC at an early stage.Biomarkers are considered novel potential targets for therapeutic intervention.It helps in the prediction of prognosis or recurrence of HCC,and also assist in the selection of appropriate treatment modality.We summarize the most relevant existing data about various biomarkers that play a key role in the progression of HCC.

Reflections on the Mechanism of Calcium Phosphate Nucleation on Titanium in Simulated Body Fluids

The results and main findings of studies reported in the literature in relation to the deposition of calcium phosphate on Ti in simulated body fluids are summarized. The effects of the surface hydroxyl groups and the sign of surface charge on the nucleation of calcium phosphate are reviewed. One major controversy among the conclusions of different studies is the order of adsorption of the calcium ions and the phosphate ions in the initial stage of immersion. A simple model based on the amphoteric nature of the hydroxyl groups on Ti is proposed in an attempt to delineate the nucleation process for calcium phosphate on Ti in simulated body fluids. HPO4^2- ions interact with the hydroxyl groups via ion exchange and/or electrostatic attraction, and Ca^2＋ ions, via electrostatic attraction only. There is no preferential order of adsorption. Seemingly inconsistent results in different studies possibly arise from different prior treatments of the samples, which affect the adsorption properties.

α-Synuclein pathology from the body to the brain:so many seeds so close to the central soil

α-Synuclein is a protein that mainly exists in the presynaptic terminals.Abnormal folding and accumulation of α-synuclein are found in several neurodegenerative diseases,including Parkinson’s disease.Aggregated and highly phospho rylated a-synuclein constitutes the main component of Lewy bodies in the brain,the pathological hallmark of Parkinson s disease.For decades,much attention has been focused on the accumulation of α-synuclein in the brain parenchyma rather than considering Parkinson s disease as a systemic disease.Recent evidence demonstrates that,at least in some patients,the initial α-synuclein pathology originates in the peripheral organs and spreads to the brain.Injection of α-synuclein preformed fibrils into the gastrointestinal tra ct trigge rs the gutto-brain propagation of α-synuclein pathology.However,whether α-synuclein pathology can occur spontaneously in peripheral organs independent of exogenous α-synuclein preformed fibrils or pathological α-synuclein leakage from the central nervous system remains under investigation.In this review,we aimed to summarize the role of peripheral α-synuclein pathology in the pathogenesis of Parkinson’s disease.We also discuss the pathways by which α-synuclein pathology spreads from the body to the brain.

Effects of different simulated fluids on anticorrosion biometallic materials

The corrosion behaviors of SUS316L stainless steel, Co Cr alloy and Ti 6Al 4V alloy in Ringer’s, PBS(-) and Hank’s solutions have been investigated. The results indicate that the corrosion of Ringer’s solution is the strongest, then followed by PBS(-) and Hank’s solution. The presence of HPO 2- 4, H 2PO - 4, SO 2- 4 and glucose in the PBS(-)and Hank’s solution probably reduces the corrosion inhibitor and corrosion current. The decrease of the solution’s pH significantly increases the corrosion rate and susceptibility to localized corrosion of SUS316L SS and Co Cr alloy. However, Ti 6Al 4V alloy exhibits an exceptional stability and has only a slight increase of corrosion rate with decreasing pH.

Raman Spectroscopy for Forensic Identification of Body Fluid Traces: Method Validation for Potential False Negatives Caused by Blood-Affecting Diseases

Two critical issues in forensic science are identifying body fluid traces found at crime scenes and preserving them for DNA analysis. However, the majority of current biochemical tests for body fluid identification, which are applicable at the crime scene, are presumptive and destructive to the sample. Raman Spectroscopy provides a suitable alternative to current methods as a nondestructive, confirmatory, and potentially in field method. Our laboratory has developed a chemometric model for the identification of five main body fluids using Raman spectroscopy. This model was developed using samples obtained from healthy donors. Thus, it is of most importance for the forensic application of the method to validate its performance for donors with diseases that might affect the biochemical composition of body fluids. In this study, the developed method was validated using peripheral blood samples acquired from donors with Celiac Disease, Sickle Cell Anemia, and Type 2 Diabetes. It was shown that the method correctly identified all samples as peripheral blood indicating that no false positives could occur because the blood traces were originated from donors suffering from the diseases.

Understanding the effects of excimer laser treatment on corrosion behavior of biodegradable Mg-1Ca alloy in simulated body fluid

In this study,a KrF excimer laser was used to modify the biodegradable Mg-1Ca alloy and the time-evolution degradation behavior of the alloy before and after laser treatment was investigated in simulated body fluid(SBF)solution using immersion tests and electrochemical impedance spectroscopy(EIS).A 5μm melted layer with a homogeneous microstructure and an MgO film on the surface were achieved by laser radiation.Corrosion observations(hydrogen evolution,morphology and corrosion products)and EIS results revealed an improvement of corrosion resistance after laser treatment for 48 h.It was found a two-layer structure developed after 2 h immersion on both the untreated and laser-treated alloys,but the sequence of forming the two layers was opposite and greatly influenced by the laser-treated layer.The time-evolution corrosion processes on the untreated and laser-treated alloys were discussed,providing a better understanding of corrosion behavior of biodegradable Mg alloys modified by excimer laser.

Corrosion fatigue of the extruded Mg-Zn-Y-Nd alloy in simulated body fluid

Magnesium alloys were considered to be used as biodegradable implants due to their biocompatibility,biodegradability and nontoxicity.However,under the simultaneous action of corrosive environment and mechanical loading in human body,magnesium alloys are easy to be affected by corrosion fatigue and stress corrosion cracking.In this work,the fatigue behavior of the extruded Mg-Zn-Y-Nd alloy used for vascular stents was studied both in air and in simulated body fluid(SBF).It was revealed that the fatigue limit of as-extruded Mg-Zn-Y-Nd alloy in air is about 65 MPa at 10^7 cycles,while there is no limit in SBF and shows a linear relationship between the fatigue life and stress amplitudes.The fatigue crack source in air was formed by the inclusions and defects.However,the stress corrosion and hydrogen embrittlement are the main reasons for the formation of the fatigue initial crack source in SBF.

Evaluation of Bioactive Properties of α and β Wollastonite Bioceramics Soaked in a Simulated Body Fluid

Dense natural wollastonite bioceramics (CaSiO3) were prepared by a sintering method, varying the pressing load and sintering temperature, in order to obtain different phases of wollastonite, and different physical properties in the materials. The products were characterized by TGA-DTA, XRD, FT-IR, SEM-EDS, TEM and XPS techniques. The results indicate the presence of two polymorphic phases of wollastonite, the β-wollastonite and α-wollastonite with a transition temperature of the β phase to α phase at approximately 1250℃. These materials were soaked in a simulated body fluid (SBF) during 1, 2 and 3 weeks, to study their solubility and bioactivity. The effect of different wollastonite phases on the solubility of Ca and Si, as well as the capacity of producing layers of “newly formed apatite” on the surfaces of these materials in SBF solution were analyzed.

Formation of Apatite in Simulated Body Fluid

It is confirmed that the essential condition for glasses and glass-ceramics to bond to living bone is the formation of an apatite layer on their surfaces in the body.It is proposed that a hydrated silica formed on the surfaces of these materials in the body plays an important role in forming the surface apatite layer,which has not been proved yet.It is shown experimentally that a pure hydrated silica gel can induce the apatite formation on its surface in a simulated body fluid when its starting pH is increased from 7.2 to 7.4.

Body Fluid Changes, Cardiovascular Deconditioning and Metabolic Impairment Are Reversed 24 Hours after a 5-Day Dry Immersion

Dry immersion is an effective and useful model for research in physiology and physiopathology. The focus of this study was to provide integrative insight into renal, endocrine, circulatory, autonomic and metabolic effects of dry immersion. We assessed if the principal changes were restored within 24 h of recovery, and determined which changes were mainly associated with immersion-induced orthostatic intolerance. Five-day dry immersion without countermeasures, and with ad libitum water intake, standardized diet and a permitted short daily rise was performed in a relatively large sample for this experiment type (14 healthy young men). Reduction of total body water derived mostly from extracellular compartment, and stabilized rapidly at the new operating point. Decrease in plasma volume was estimated at 20% - 25%. Five-day immersion was sufficient to impair metabolism with a decrease in glucose tolerance and hypercholesterolemia, but was not associated with pronounced autonomic changes. Five-day immersion induced marked cardiovascular impairment. Immediately after immersion, over half of the subjects were unable to accomplish the 20-min 70° tilt;during tilt, heart rate and total peripheral resistance were increased, and stroke volume was decreased. However, 24 hours of normal physical activity appeared sufficient to reverse orthostatic tolerance and all signs of cardiovascular impairment, and to restitute plasma volume and extracellular fluid volume. Similarly, metabolic impairment was restored. In our study, the major factor responsible for orthostatic intolerance appeared to be hypovolemia. The absence of pronounced autonomic dysfunction might be explained by relatively short duration of dry immersion and daily short-time orthostatic stimulation.

Effects of geosorbent and solution properties on sorption and desorption of PAHs

Characteristics of phenanthrene and pyrene’s sorption and desorption on two local soils in solutions of simulated groundwater,simulated lung fluid,and simulated saliva were studied with batch equilibrium experiments to understand the fate of PAHs in the karst region of southwestern China and to assess the environmental exposure and the health risk of PAHs.The results showed that the sorption and desorption isotherms of phenanthrene and pyrene on two target soils in the three solution systems could be adequately described by the Freundlich model,while the fitted isotherm parameters for the simulated groundwater solution distinguished notably from those for the simulated body fluid solutions.For the sorption experiments,in the simulated groundwater,the n values were 0.722 and 0.672 for phenanthrene and were 0.724 and0.663 for pyrene,respectively,on the yellow soil and the limestone soil;The log KF values were 3.118 and 3.323 for phenanthrene and were 3.648 and 3.846 for pyrene,respectively,on the yellow soil and the limestone soil.In the simulated body fluids,the n values for phenanthrene and pyrene ranged from 0.622 to 0.836 and from 0.590 to0.865,respectively,and the log KF values of phenanthrene and pyrene ranged from 2.845 to 3.327 and from 3.344 to3.779,respectively.For the desorption experiments,in the simulated groundwater,the n values were 0.662 and 0.744 for phenanthrene and were 0.702 and 0.647 for pyrene,respectively,on the yellow soil and the limestone soil.The log KF values were 3.666 and 3.686 for phenanthrene and were 4.128 and 4.225 for pyrene,respectively,on the yellow soil and the limestone soil.In the simulated body fluids,the n values for phenanthrene and pyrene ranged from 0.612 to 0.668 and from 0.631 to 0.819,respectively,and the log KF values of phenanthrene and pyrene ranged from 3.134 to 3.407 and from 3.533 to 3.839,respectively.The limestone soil had relatively higher log KF values but lower KOC values compared to those of the yellow soil,indicated that the nature of sorbent soils played the dominant role in sorption and desorption behaviors of PAHs.The experimental results showed a remarkable differences in sorption and desorption behaviors of PAHs in simulated body fluids and groundwater.The nonlinearities of measured isotherms and the measured sorption capacities of soils in simulated body fluids were significantly lower than corresponding those in the simulated groundwater,and HI values for simulated body fluids systems were significantly smaller than corresponding those for the simulated groundwater systems.The results underscore cautions in assessing environmental exposure and health risks of PAHs based on their sorption-desorption data in simulated groundwater as this is traditionally done.

Manifestation of Pathological States of Numerous Diseases in the Largest Organ of the Human Body: (II) From Pancreatitis to Pancreatic Cancer Invasion, Formation of Stroma around the Primary Tumor in the Fascia, to Early Detection of Non-Coding microRNAs in Body Fluids and Development of Drugs to Treat Different Stages of Pancreatic Cancer

Patients suffering from pancreatic ductal adenocarcinoma (PDAC) have an average survival time of 4 - 6 months after confirmed diagnosis. The primary tumor is surrounded by a thick interstitial fluid with high pressure and dense distribution of collagen, forming a huge stroma, rendering the tumor resistant to chemo- and radiotherapy. From the genetic point of view, pancreatic carcinogenesis is driven by mutations, resulting in common activation of the oncogene KRAS, and/or inactivation of one or more of the tumor suppressor genes CDKN2A, TP53, SMAD4 [1]. The pancreas is a mixed exocrine and autocrine organ, with different cell types building up the organ. The pathogenesis involves more than 13 signaling pathways at different stages. Off-balance of the function of the proteins in these pathways due to the stated 4 plus other mutations could readily lead to carcinogenesis. We first present the basic mechanism of these 13 relevant pathways. We then provide a detailed analysis of the progression of this disease, from pancreatitis to tumor formation and metastasis, with special attention on the roles played by the newly discover calcium channel Piezo, stellate cells, stem-cell-like cells, and the concept invadopodium. Thirty potential drugs, based on in vitro and xenograft experiments from different groups, are discussed, including vitamins A, Tocotrienols-E, and D, chemical compounds, non-coding micro RNAs, circular RNA, piwi-interacting RNAs. The recent detection of exosomes enclosing many of these RNAs in body fluids gives us hope of developing early detection methodology because these RNAs carry messages for cell-cell communication at a distance. Delivery of potent drugs by nanoparticles gives us chance to send drugs through the stroma to target the tumor. Since body fluids form a circulating system, together with the connective tissues (where the tumor is associated) form the largest organ—the fascia, we conclude that manifestation of successive pathological states of pancreatic carcinogenesis can be found in compartments of the fascia. We present 17 figures, hoping to ease off the complexity of the pathogenesis of this most lethal cancer disease.

Super-hydrophobic stearic acid layer formed on anodized high purified magnesium for improving corrosion resistance of bioabsorbable implants

Magnesium and its alloys are ideal candidates for bioabsorbable implants.However,they can dissolve too rapidly in the human body for most applications.In this research,high purified magnesium(HP-Mg)was coated with stearic acid(SA)to slow the corrosion rate of magnesium in simulated body fluid at 37±1°C.HP-Mg was anodized(AC and DC voltages)to form an oxide/hydroxide layer,and then it was immersed in a SA solution.The SA coated layer surface,anodized layer,and the thickness of the oxide/hydroxide layer were investigated with Scanning Electron Microscopy(SEM).Electrochemical impedance spectroscopy(EIS)and potentiodynamic polarization(PDP)were used to estimate the corrosion rate of HP-Mg specimens.The results confirm that the hydrophobic coating can decrease the corrosion rate of HP-Mg by more than 1000x.The protectiveness of coated layer for anodized specimens with AC voltage continue for 2 and 3 weeks.For the HP-Mg coated anodized with DC voltage,the coated layer could improve the corrosion resistance for only a few days.

Physical and degradation properties of PLGA scaffolds fabricated by salt fusion technique

Tissue engineering scaffolds require a controlled pore size and interconnected pore structures to support the host tissue growth. In the present study, three dimensional （3D） hybrid scaffolds of poly lactic acid （PLA） and poly glycolic acid （PGA） were fabricated using solvent casting/particulate leaching. In this case, partially fused NaCl particles were used as porogen （200-300μ） to improve the overall porosity （≥90%） and internal texture of scaffolds. Differential scanning calorimeter （DSC） analysis of these porous scaffolds revealed a gradual reduction in glass transition temperature （Tg） （from 48°C to 42.5°C） with increase in hydrophilic PGA content. The potential applications of these scaffolds as implants were further tested for their biocompatibility and biodegradability in four simulated body fluid （SBF） types in vitro. Whereas, simulated body fluid （SBF） Type1 with the optimal amount of HCO 3 ions was found to be more appropriate and sensible for testing the bioactivity of scaffolds. Among three combinations of polymer scaffolds, sample B with a ratio of 75：25 of PLA： PGA showed greater stability in body fluids （pH 7.2） with an optimum degradation rate （9% to 12% approx）. X-ray diffractogram also confirmed a thin layer of hydroxyapatite deposition over sample B with all SBF types in vitro.

Development and assessment of a multifunctional chitosan-based coating applied on AZ31 magnesium alloy: Corrosion resistance and antibacterial performance against Klebsiella Pneumoniae

This work presents a simple method to functionalise the surface of AZ31 magnesium alloy by applying a duplex MgF_(2)/chitosan coating,which improves its corrosion resistance and provides it with some antibacterial performance.First,the effect of three chitosan solutions with different concentrations on the growth of the bacteria Klebsiella pneumoniae in nutritive medium(TSB)was evaluated by absorbance kinetics experiments,where the chitosan solution at 2%(m/V)was selected for the coating preparation.Before coating application,the AZ31 substrate was pretreated with hydrofluoric acid for 48 hours in order to form a MgF_(2)conversion layer.Subsequently,the coating was applied to the pretreated substrate through the dry-casting method.Samples of the alloy in each surface condition(bare,pretreated,and pretreated+coated with chitosan)were exposed to simulated body fluid(SBF)for 21 days at 37°C,with the solution renewed every 24 hours and the wastes stored.The surfaces were characterised by SEM-EDS,and XPS after the immersion tests,whereas the stored solutions were employed to measure the change in the Mg-ions concentration.Electrochemical impedance spectroscopy and potentiodynamic polarisation were performed in each surface condition to compare their corrosion resistance in SBF.The antibacterial activity of the functionalised surfaces was evaluated by the plate counting method and compared with bare samples.All results were correlated and demonstrate that the modified surface of AZ31 achieved a higher corrosion resistance when it was exposed to SBF,as well as a reduction of the bacterial growth during in vitro tests.

Gallium-containing magnesium alloy for potential use as temporary implants in osteosynthesis

In recent years magnesium alloys have been studied intensively with a view to their potential use in bioresorbable medical implants.In the present work the microstructure and the corrosion properties of a new bioresorbable Mg-4 wt%Ga-4 wt%Zn alloy and its variants with low Ca,Nd or Y additions were investigated.These alloys are of interest due to the efficacy of gallium as an element inhibiting bone resorption,osteoporosis,Paget's disease,and other illnesses.A severe plastic deformation technique of equal channel angular pressing(ECAP)was shown to provide the alloys with favorable mechanical properties.In addition,a desirable low rate of degradation in a simulated body fluid(Hanks'solution)was achieved.

Sol-gel Derived TiO_2-Bioactive glass-Hydroxyapatite Bioactive Coating on Titanium Alloy Substrate

Coating the hydroxyapatite （HA） on the titanium alloy surface can obtain a bioactive implant with high mechanical properties However, the bonding force between the titanium alloy and the HA was low due to their different coefficient of thermal expansion （CET）. Preparing the multi-layer coating with alleviated thermal stress on titanium alloy substrate is few reported. Fabrication of a TiO2-bioactive glass （BG）-HA bioactive coating was proposed to solve this problem. A particular TiO2 surface was prepared on the titanium alloy substrate by micro-arc oxidation treatment. The BG and HA coating were coated onto the TiO2 surface in turn by using a sol-gel method. The microstructure, surface morphology and phase composition of the coatings were analyzed. The bonding force of coatings was investigated by the nick apparatus. In vitro dissolution was performed by soaking the TiO2-BG-HA coated samples into the simulated body fluid for various periods. Micro-structural observations indicated that no delamination and crack occurred at the interface of HA/BG and BG/TiO2. The bonding between the substrate and coating consists of the mechanical interaction and the chemical bonding. The bonding force could reach about 45 N. The TiO2-BG-HA coating displayed the excellent forming ability of bone-like apatite when it was soaked into the simulated body fluid. This work suggests an innovative way to reduce the internal stress among coatings through varying BG composition to adjust its CTE, so as to enhance the bonding force.

Coupled Aerodynamic and Hydrodynamic Analysis of Floating Offshore Wind Turbine Using CFD Method

To simulate floating offshore wind turbine(FOWT)in coupled wind-wave domain via CFD method,the NREL 5MW wind turbine supported by the OC3-Hywind Spar platform is modeled in the STAR-CCM+ software.Based on the Reynolds-averaged Navier-Stokes(RANS)equations and re-normalisation group(RNG)k-εturbulence model,the rotor aerodynamic simulation for wind turbine is conducted.Numerical results agree well with the NREL data.Taking advantage with the volume of fluid(VOF)method and dynamic fluid body interaction(DFBI)technology,the dynamic responses of the floating system with mooring lines are simulated under the coupled wind-wave sea condition.The free-decay tests for rigid-body degrees of freedom(DOFs)in still water and hydrodynamic tests in a regular wave are performed to validate the numerical model by comparing its result with the results simulated by FAST.Finally,the simulations of the overall FOWT system in the coupled wind-wave flow field are carried out.The relationship between the power output and dynamic motion responses of the platform is investigated.The numerical results show that the dynamic response of wind turbine performance and platform motions all vary in the same frequency as the inlet wave.During platform motion,the power output of wind turbine is more sensitive than the thrust force.This study may provide some reference for further research in the coupled aero-hydro simulation of FOWT.

Effects of tap water, electrolyte solution, and spontaneous and furosemide-stimulated urinary excretion on thirst

AIM: To contrast the effects of various modifications of body fluid volumes on thirst as reported by healthy volunteers. METHODS: Ten male volunteers aged between 19 and 37 years(mean 22 years) underwent four experiments each, which comprised infusion of 400-800 m L of acetated Ringer's solution and intake of 600 m L of tap water. Half of the experiments were preceded by volume depletion(median 1.7 L) with furosemide. A visual analogue scale(0-100 mm) was used to assess perceived thirst during each experiment. RESULTS: Volume depletion(P < 0.001) and tap water(P < 0.03) both affected thirst by 13 mm per L of fluid, whereas spontaneous diuresis and infusion of Ringer's acetate did not significantly change the thirst rating(multiple regressions). More detailed analyses showed that the volume depletion increased the median(25th-75th percentiles) thirst rating from 28 mm(21-43) to 59 mm(46-72, P < 0.001) while no change occurred in those who were only slightly thirsty(< 30 mm) before the volume depletion began. Ringer's solution alleviated thirst in those who were very thirsty, but tended to increase thirst in the volunteers who were not thirsty before the infusion. Similarly, hydration with tap water decreased thirst(by 24 mm, P < 0.04) in those who were thirsty(> 60 mm) while the others reported no change.