Dilution Ratio Research Articles

Establishment and evaluation of a method for extracting exogenous short DNA fragments of Schistosoma japonicum from urine samples

To establish the method for extracting exogenous short DNA fragments of Schistosoma japonicum from urine samples, and to evaluate the efficiency of this method for extraction from urine samples treated with various methods. The S. japonicum SjG28 gene fragment was selected as a target sequence, and the 81 bp short DNA fragment was amplified on the target sequence using PCR assay. Following characterization using sequencing, the short DNA fragment was added into the urine samples as an exogenous short DNA fragment. Primers and probes were designed with SjG28 as a target gene, to establish the real-time fluorescent quantitative PCR (qPCR) assay. The sensitivity of this qPCR assay was evaluated with exogenous short DNA fragments that were diluted at a 1:10 dilution ratio as the DNA template, and the specificity of the qPCR assay was evaluated with the genomic DNA of S. mansoni, S. haematobium, Babesia, Ancyiostoma duodenaie, Cionorchis sinensis, and Paragonimus westermani as DNA templates. Exogenous short DNA fragments were added into artificial and healthy volunteers' urine samples, followed by pH adjustment, centrifugation and concentration, and the efficiency of extracting exogenous short DNA fragments from urine samples was compared with the QIAmp Viral RNA Mini Kit (Qiagen kit) and BIOG cfDNA easy kit (BIOG kit). An 81 bp small DNA fragment of S. japonicum was successfully prepared, and the lowest detection limit of the established qPCR assay was 100 copies/μL of the 81 bp small DNA fragment of S. japonicum. If the genomic DNA of S. japonicum, S. mansoni, S. haematobium, Babesia, A. duodenaie, C. sinensis, and P. westermani served as DNA templates, the qPCR assay only detected fluorescent signals with S. japonicum genomic DNA as the DNA template. If the pH values of artificial urine samples were adjusted to 5, 6, 7 and 8, the recovery rates were (49.12 ± 2.09)%, (84.52 ± 4.96)%, (89.38 ± 3.32)% and (87.82 ± 3.90)% for extracting the exogenous short DNA fragment of S. japonicum with the Qiagen kit, and were (2.30 ± 0.07)%, (8.11% ± 0.26)%, (13.35 ± 0.61)% and (20.82 ± 0.68)% with the BIOG kit, respectively (t = 38.702, 26.955, 39.042 and 29.571; all P values < 0.01). If the Qiagen kit was used for extracting the exogenous short DNA fragment from artificial urine samples, the lowest recovery rate was seen from urine samples with a pH value of 5 (all P values < 0.05), and there were no significant differences in the recovery rate from urine samples with pH values of 6, 7 and 8 (all P values > 0.05). Following centrifugation of artificial [(64.30 ± 1.00)% vs. (58.87 ± 0.26)%; t = 12.033, P < 0.05] and healthy volunteers' urine samples [(31 165 ± 1 017) copies/μL vs. (28 471 ± 818) copies/μL; t = 23.164, P < 0.05]. In addition, concentration of artificial urine samples with the 10 kDa Centrifugal Filter and concentration of healthy volunteers' urine samples with the 100 kDa Centrifugal Filter were both effective to increase the recovery of the Qiagen kit for extracting the exogenous short DNA fragment of S. japonicum (both P values < 0.01). A method for extracting exogenous short DNA fragments of S. japonicum from urine samples has been successfully established, and the Qiagen kit has a high extraction efficiency. Adjustment of urine pH to 6 to 8 and concentration of healthy volunteers' urine samples with the 100 kDa Centrifugal Filter are both effective to increase the efficiency of extracting exogenous short DNA fragments of S. japonicum.

Comparative studies on microstructure and properties of CoCrFeMnNi high entropy alloy coatings fabricated by high-speed laser cladding and normal laser cladding

Two kinds of CoCrFeMnNi high entropy alloy (HEA) coatings were prepared by high-speed laser cladding (HS-) and normal laser cladding (N-), respectively. The phase, microstructure, micromechanical properties, and corrosion resistance of both coatings were analyzed. According to the results, both coatings are stable single-phase FCC structures with the dendrites microstructure. However, the dendrites of the HS-CoCrFeMnNi coating are finer. Owing to the low heat input and the quick cooling and heating characteristics of high-speed laser cladding, the width of the heat affected zone and the dilution ratio of the HS-CoCrFeMnNi coating are reduced. In addition, the microhardness of the HS-CoCrFeMnNi coating (∼ 184 HV0.2) is higher than that of the N-CoCrFeMnNi coating (∼ 160 HV0.2). The elastic modulus (Er) of the HS-CoCrFeMnNi coating is significantly increased in order to enhance its resistance to plastic deformation. The plastic storage energy Qp of the HS-CoCrFeMnNi coating is higher, revealing the improved crack resistance. The self-corrosion potential Ecorr of the HS-CoCrFeMnNi coating positively shifted by 0.142 V compared with the N-CoCrFeMnNi coating, and the self-corrosion current density Icorr decreased from 6.3 × 10−6 A/cm2 to 2.9 × 10−7 A/cm2 in 3.5% NaCl solution. Therefore, the HS-CoCrFeMnNi coating possesses better corrosion resistance and comprehensive mechanical properties.

Inactivation Performance of Pseudorabies Virus as African Swine Fever Virus Surrogate by Four Commercialized Disinfectants

This study was based on similar physicochemical characteristics of pseudorabies virus (PRV) and African swine fever virus (ASFV). A cellular model for evaluation of disinfectants was established with PRV as an alternative marker strain. In the present study, we evaluated the disinfection performance of commonly used commercialized disinfectants on PRV to provide a reference for the selection of good ASFV disinfectants. In addition, the disinfection (anti-virus) performances for four disinfectants were investigated based on the minimum effective concentration, onset time, action time, and operating temperature. Our results demonstrated that glutaraldehyde decamethylammonium bromide solution, peracetic acid solution, sodium dichloroisocyanurate, and povidone-iodine solution effectively inactivated PRV at concentrations 0.1, 0.5, 0.5, and 2.5 g/L on different time points 30, 5, 10, and 10 min, respectively. Specifically, peracetic acid exhibits optimized overall performance. Glutaraldehyde decamethylammonium bromide is cost effective but requires a long action time and the disinfectant activity is severely affected by low temperatures. Furthermore, povidone-iodine rapidly inactivates the virus and is not affected by environmental temperature, but its application is limited by a poor dilution ratio such as for local disinfection of the skin. This study provides a reference for the selection of disinfectants for ASFV.

An investigation on graphite behavior and coating properties in the molten pool based on different powder particle sizes

This paper aims to explore the influence mechanism of powder particle sizes on the microstructure and properties of coatings and identify the effect of powder particle size difference on the coating graphite phase. NbC-reinforced Ni-based coatings were in-situ synthesized by laser cladding to investigate the impact of powder particle sizes on the morphology, structure, and properties of coatings. The results indicate that increasing powder particle size enlarges the coating area and decreases the coating width and dilution ratio. Meanwhile, the defect ratio first increases and then decreases. The XRD test suggests that the coating mainly consists of NbC, solid solution (Fe, Ni), B4C, Cr2C, and CrB2. Different powder particle sizes do not change the phase composition of coatings but affect the graphite phase morphology. The morphology transforms from spherical to flocculent as the powder size varies from micrometer to nanometer. The hardness of coatings gradually increases, and the friction and wear properties decrease with the growth of powder particle size. The dispersed graphite phase in the nano coating plays a self-lubricating role in the friction and wear process. This research provides a reference and theoretical basis for selecting powder particle size in laser cladding.

Study on the Process Characteristics Based on Joule Heat of Sliding-Pressure Additive Manufacturing

This study developed an experimental system based on Joule heat of sliding-pressure additive manufacturing (SP-JHAM), and Joule heat was used for the first time to accomplish high-quality single-layer printing. The roller wire substrate is short-circuited, and Joule heat is generated to melt the wire when the current passes through. Through the self-lapping experimental platform, single-factor experiments were designed to study the effects of power supply current, electrode pressure, contact length on the surface morphology and cross-section geometric characteristics of the single-pass printing layer. Through the Taguchi method, the effect of various factors was analyzed, the optimal process parameters were obtained, and the quality was detected. The results show that with the current increase, the aspect ratio and dilution rate of a printing layer increase within a given range of process parameters. In addition, with the increase in pressure and contact length, the aspect ratio and dilution ratio decrease. Pressure has the greatest effect on the aspect ratio and dilution ratio, followed by current and contact length. When a current of 260 A, a pressure of 0.60 N and a contact length of 1.3 mm are applied, a single track with a good appearance, whose surface roughness Ra is 3.896 μm, can be printed. Additionally, the wire and the substrate are completely metallurgically bonded with this condition. There are also no defects such as air holes and cracks. This study verified the feasibility of SP-JHAM as a new additive manufacturing strategy with high quality and low cost, and provided a reference for developing additive manufacturing technology based on Joule heat.

Spermatozoa Cryopreservation of Sex-Reversed Rainbow Trout (Oncorhynchus mykiss): The Effect of Dilution Rate and Supplementation of a N-(2-Mercaptopropionyl)-Glycine -Based Extender on Sperm Motility and Fertilizing Capacity

For commercial aquaculture, the obtainment of all-female salmonid populations is important in fish farms. After freezing process, variable spermatozoa maturity, higher sperm concentration, low sperm quality, and reduced fertilization success have been observed in sex-reversed female rainbow trout. For these reasons, the objective of this study was to assess effect of dilution rate and supplementation of a N-(2-Mercaptopropionyl)-Glycine (MPG)-based extender on sperm motility and fertilizing capacity of sex-reversed rainbow trout (Oncorhynchus mykiss). The supplementation of MPG [0 (control, 0), 1 mM; 2 mM; 4 mM] to extenders and dilution ratio (1:9, 1:15 and 1:25) were tested in sex-reversed female rainbow trout spermatozoa during cryopreservation. For thawing, the straws (0.5 ml) were placed in a water bath at 37°C for 30 s. Our results showed that the best concentration of MPG was 2 mM for post-thaw motility duration (120.67±9.07%), fertilization (62.67±3.10%) and hatching rate (54.33±3.10%) at 1:15 dilution rate. Overall, MPG provided improvements during cryopreservation process and could be used as protective agent.

Comparative of Forensic DNA Identification Using Cell Lysis Method and Magnetic Beads Method.

To compare the effects of cell lysis method and magnetic beads method in forensic DNA identification and to explore these two methods in forensic DNA identification. The genome DNA of THP-1 cells in different quantities was extracted by the cell lysis method and magnetic beads method, and the DNA content was quantified by real-time quantitative PCR. The cell lysis method and magnetic beads method were used to type the STR of human blood with different dilution ratios. When the numbers of THP-1 cell were 100, 400 and 800, the DNA content extracted by cell lysis method were (1.219±0.334), (5.081±0.335), (9.332±0.318) ng, respectively; and the DNA content extracted by magnetic beads method were (1.020±0.281), (3.634±0.482), (7.896±0.759) ng, respectively. When the numbers of THP-1 cells were 400 and 800, the DNA content extracted by the cell lysis method was higher than that by the magnetic beads method. The sensitivity of cell lysis method and magnetic beads method was similar in STR typing of human blood at different dilution ratios. Complete STR typing could be obtained at 100, 300 and 500-fold dilutions of blood samples, but could not be detected at 700-fold dilution. STR typing of undiluted human blood could not be detected by cell lysis method. The cell lysis method is easy to operate and can retain template DNA to the maximum extend. It is expected to be suitable for trace blood evidence tests.

Laser cladding of IN713 LC superalloy with Amdry 997 powder: microstructural evolution

In this research, Amdry 997 powder was applied by a laser cladding process to the IN713 Low Carbon (IN713 LC) nickel-based superalloy. To evaluate the microstructural behaviors of the clad zones, different variables (the laser speed, laser frequency, and pulse width) were used in the laser cladding process. The final aim of this study was to achieve a clad zone without obvious cracks and pores with a low dilution ratio. The results showed that the upper region of the cladding includes very small equiaxed dendrites. The middle region of the cladding includes columnar dendrites. Equiaxed and columnar grains can be formed at the interface region. With an increase in the heat input of the laser (increasing the pulse width and laser frequency or decreasing the laser speed), the solidification rate reduces and consequently, a longer solidification time exists. Hence, more elongated grains are formed at the interface region. The microstructure of coatings in the upper and middle regions includes γ-Ni, β-NiAl and γ′-Ni3Al phases and TaC carbide, whereas the microstructure of the coatings at the interface region includes γ and β-NiAl and γ′-Ni3Al phases and M (Mo, Ta)C carbide. The optimum laser parameters that result in a low dilution ratio (14%) and a dense clad zone without obvious crack and porosity are a laser speed of 140 mm min−1,laser frequency of 14 Hz, and pulse width of 9 ms.

Optimization of the Additive Manufacturing of Co-Cr Alloy by using DED Process

In this study, the directed energy deposition (DED) process was applied to additively manufacture cobalt-chromium alloy materials, which are commonly used in molds, dental/orthopedic medical applications, vehicles, and aircraft. The quality of the deposition obtained by using the DED technology is affected by process parameters. Therefore, the aim of this experiment, was to evaluate the microstructure changes caused by varying parameters such as the laser power, laser header moving speed, and powder feeding rate, with the goal of improving and optimizing the stacking quality. The shape, heat-affected zone, and dilution ratio of the microstructure were analyzed using an optical microscope. Furthermore, the global energy density(GED) was calculated because it affects the quality of the 3D-printed product. The optimal process conditions for the DED method determined by calculating the GED under various process conditions.

Impact of different brushing/abrasion protocols on erosive tooth wear for in vitro studies

ObjectiveTo investigate the influence of different toothbrushing (with dentifrice) protocols on the progression of erosive tooth wear for in vitro studies. DesignBovine enamel specimens were randomly distributed into 12 experimental groups (n = 10), according to the study factors: (1) brushing movement (horizontal or circular); (2) slurry diluent (artificial saliva or distilled water); (3) toothpaste dilution ratio (1:2, 1:3 or 1:4). A 5-day erosion-abrasion cycling model was performed, each consisting of 4 erosive challenges (0.3 % citric acid, pH=2.6) followed by 60 min exposure to artificial saliva. Brushing with fluoride toothpaste (15 s, 1400 ppm F-, AmF) was carried out 2x/day. Enamel surface loss (SL) was determined by optical profilometry. Data were statistically analyzed with three-way ANOVA and Tukey tests (α = 0.05). ResultsSL was lower for the horizontal movement than for the circular (p = 0.044). There were no significant differences among the dilution ratios for artificial saliva. For distilled water, the more concentrated slurry (1:2) presented greater surface loss than the less concentrated slurries (1:3 and 1:4, p = 0.049 and p = 0.014, respectively). Dilutions with artificial saliva at ratios 1:3 and 1:4 presented higher surface loss than with distilled water (p = 0.008 and p < 0.001, respectively); however, for 1:2 ratio, there were no significant differences between the diluents. ConclusionsThe in vitro progression of enamel SL was influenced by the brushing movement, as well as the combination of the dilution ratio and the diluent of the toothpaste slurry, and therefore, all these factors must be considered when comparing results from different studies.

Establishment and Application of an Indirect ELISA for the Detection of Antibodies to Porcine Streptococcus suis Based on a Recombinant GMD Protein

S. suis is an important zoonotic pathogen from sick and recessive carrier pigs that poses a serious threat to animal husbandry production and public health. It usually causes horizontal transmission among pigs. The morbidity and mortality of this disease are very high. Human infection is caused through direct or indirect contact with sick pigs. The two large-scale outbreaks in China were due to the outbreak of S. suis on pig farms, which spread to human infection; thus, detecting S. suis in pig herds is crucial. At present, the commercial S. suis ELISA type 2 kits on the market can only detect single serotypes, high probabilities of interaction reactions, and biosafety risks when using inactivated S. suis as an antigen. Phosphate-3-glyceraldehyde dehydrogenase (GAPDH), muramidase-released protein (MRP), and dihydrolipoamide dehydrogenase (DLDH) are important S. suis type 2, S. suis type 7, and S. suis type 9 protective antigens. This study purified the GMD protein (B-cell-dominant epitopes of GAPDH, MRP, and DLDH antigens) and used a diverse combination of dominant epitopes of the multiple different antigens as coated antigens, improving the sensitivity and safety of the indirect ELISA experiments. An indirect ELISA method (GMD-ELISA) was developed for detecting S. suis antibodies. The antigen-antibody response was optimized using checkerboard titration. The results of testing using ELISA for Salmonella enterica (S. enterica), Escherichia coli (E. coli), Staphylococcus aureus (SA), and Streptococcus pyogenes (S. pyogenes) were all negative, indicating that this method had strong specificity. The results were still positive when the dilution ratio of S. suis-positive serum reached 1:6, 400, thus indicating that the method had high sensitivity. The results of the reproducibility assay for indirect ELISA showed that the intra-assay coefficient of variation and the inter-assay coefficient of variation were less than 10%, indicating that the method had good repeatability. We investigated the seroprevalence of S. suis in 167 serum samples collected in East China, and 33.5% of the samples were positive for antibodies against S. suis, indicating that the prevalence of S. suis is high in pig farms in Eastern China. The novel GMD-ELISA is a convenient, sensitive, and specific diagnostic method that provides technical support for rapid diagnosis and epidemiological investigation.

Construction and Manipulation of Serial Gradient Dilution Array on a Microfluidic Slipchip for Screening and Characterizing Inhibitors against Human Pancreatic Lipase.

Obesity is one of the foremost public health concerns. Human pancreatic lipase (hPL), a crucial digestive enzyme responsible for the digestion of dietary lipids in humans, has been validated as an important therapeutic target for preventing and treating obesity. The serial dilution technique is commonly used to generate solutions with different concentrations and can be easily modified for drug screening. Conventional serial gradient dilution is often performed with tedious multiple manual pipetting steps, where it is difficult to precisely control fluidic volumes at low microliter levels. Herein, we presented a microfluidic SlipChip that enabled formation and manipulation of serial dilution array in an instrument-free manner. With simple slipping steps, the compound solution could be diluted to seven gradients with the dilution ratio of 1:1 and co-incubated with the enzyme (hPL)-substrate system for screening the anti-hPL potentials. To ensure complete mixing of solution and diluent during continuous dilution, we established a numerical simulation model and conducted an ink mixing experiment to determine the mixing time. Furthermore, we also demonstrated the serial dilution ability of the proposed SlipChip using standard fluorescent dye. As a proof of concept, we tested this microfluidic SlipChip using one marketed anti-obesity drug (Orlistat) and two natural products (1,2,3,4,6-penta-O-galloyl-β-D-glucopyranose (PGG) and sciadopitysin) with anti-hPL potentials. The IC50 values of these agents were calculated as 11.69 nM, 8.22 nM and 0.80 μM, for Orlistat, PGG and sciadopitysin, respectively, which were consistent with the results obtained by conventional biochemical assay.

Distribution of environmentally persistent free radicals in size-segregated PMs emitted from residential biomass fuel combustion

Environmentally persistent free radicals (EPFRs) are considered as an emerging pollutant due to their potential environmental risks, but the distribution characteristics of particulate matters (PMs)-EPFRs from residential combustion source are poorly understood. In this study, biomass (corn straw, rice straw, pine and jujube wood) combustion was studied in lab-controlled experiments. More than 80% of PM-EPFRs were distributed in PMs with aerodynamic diameter (dae) ≤ 2.1 µm, and their concentration in fine PMs was about 10 times that in coarse PM (2.1 µm ≤ dae ≤ 10 µm). The detected EPFRs were carbon-centered free radicals adjacent to oxygen atoms or a mixture of oxygen- and carbon-centered radicals. The concentrations of EPFRs in coarse and fine PMs were positively correlated with char-EC, but the EPFRs in fine PMs exhibited a negative correlation with soot-EC (p < 0.05). The increase of PM-EPFRs signals with the increased dilution ratio during pine wood combustion was more significant than that from rice straw, which may be resulted from the interactions between the condensable volatiles and the transition metals. Our study provides useful information for better understanding the formation of combustion-derived PM-EPFRs, and will be instructive for its purposeful emissions control.

Effect of pretreatment approach on the ELISA-based detection of cyanotoxins in water: Analysis and application

Common cyanotoxins, such as microcystins and nodularins, are produced by frequently occurring harmful cyanobacterial algal blooms in freshwater systems. The required routine monitoring of microcystins and nodularins in drinking water and ambient water demands cost-efficient and reliable enzyme-linked immunosorbent assay kits. We validated the performance of a self-produced broad-spectrum enzyme-linked immunosorbent assay kit and investigated two different methods of mitigating the matrix effects to elucidate the effect of the respective pretreatment approaches recommended by China and the United States on the quantitative detection of cyanotoxins in surface water. We found that the enzyme-linked immunosorbent assay kit achieved a detection limit of 0.15 μg/L with a linear detection range from 0.27 μg/L to 1.87 μg/L for microcystin-LR (the most studied and widely distributed cyanotoxin). The matrix effects could be mitigated both by dilution of water samples with an optimal dilution ratio and dilution of antibody with the buffer containing phosphate buffer solution (10×), bovine serum albumin (1 %) and ethylene diamine tetraacetic acid (0.5 %). In terms of the surface water samples being tested, the concentrations of microcystins and nodularins measured based on pretreatment approach recommended by the United States were 1– 5 times that measured based on pretreatment approach recommended by China, indicating that the pretreatment approach of China overlooks cyanotoxins. In addition, all the measured total microcystins and nodularins of the surface water samples were below the health advisory limit (1.6 μg/L) for microcystins in drinking water proposed by the United States Environmental Protection Agency for school-age children and adults. Our research could provide significant information for outbreak warnings and risk management of harmful cyanobacterial algal blooms.

Comprehensive influence of uncertainty propagation of chemical kinetic parameters on laminar flame speed prediction: a case study of dimethyl ether

The uncertainties existing in the parameters of chemical kinetic models have a non-negligible influence on the model predictions. It is necessary to conduct a quantitative uncertainty analysis to explore the influence of each parameter on chemical mechanism predictions. To comprehensively consider the effect of the uncertainties of reaction rate parameters, thermodynamic parameters, and transport parameters on model predictions, local sensitivity analysis, local-sensitivity-based uncertainty analysis (LSUA), and random-sampling high dimensional model representation (RS-HDMR) method were coupled to investigate the uncertainty propagation of the chemical kinetic parameters to the calculated laminar flame speed of dimethyl ether under a wide range of conditions using a detailed mechanism. First, the uncertainty analysis was conducted using the local sensitivity analysis and the LSUA method under a wide range of operating conditions to identify the important operating conditions and chemical kinetic parameters. It is found that the prediction uncertainty of laminar flame speed is more obvious under the conditions of high dilution ratio, high pressure, and large equivalence ratio than that under other conditions. According to the results of LSUA, the prediction uncertainty is mainly from the reaction rate coefficients and thermodynamic data. Then, the uncertainty propagation from the significant parameters to the calculated laminar flame speed under important conditions was analysed using the RS-HDMR method. To reduce the huge computational cost of the RS-HDMR method, the backpropagation artificial neural network was employed. The RS-HDMR results indicate that the reaction H + O2 = O + OH has the highest sensitivity coefficient under the whole investigated conditions, which is different from the results using the LSUA method. The non-linear relationship between the rate coefficient and the predicted laminar flame speed is responsible for the discrepancy. Furthermore, it is found that the sensitivity coefficient of the input parameters strongly depends on the operating conditions.

Antibody avidity to pertussis toxin after acellular pertussis vaccination and infection

ABSTRACT Pertussis toxin (PT) is a unique virulence factor of Bordetella pertussis, and therefore a key component of acellular pertussis vaccines. Although immunity after infection seems to persist longer than after vaccination, the exact mechanisms are not fully known. In this study the overall binding strength (avidity) of anti-PT IgG antibodies was compared after acellular booster vaccination and infection, as a parameter to evaluate long-lasting protection. Danish and Finnish serum samples from a total of 134 serologically confirmed patients and 112 children who received acellular booster vaccines were included in this study. The concentration of anti-PT IgG was first determined by ELISA, followed by two separate ELISAs to evaluate antibody avidity: either with a dilution series of urea as a bond-breaking agent of antibody and antigen binding and a constant anti-PT IgG concentration between the samples or with a constant dilution ratio of sera and detergent. In addition to urea, the use of diethylamine and ammonium thiocyanate as disruptive agents were first compared between each other. A strong Spearman correlation (R > 0.801) was noted between avidity and concentration of anti-PT IgG antibodies if a constant serum dilution method was used, and avidity was noted to be higher in patients in comparison to vaccinees in Denmark, but not in Finland. However, no correlation between antibody concentration and avidity was found if a constant anti-PT IgG concentration was used (R = −0.157). With this method, avidity after vaccination was significantly higher in comparison to that after infection in both Danish and Finnish subjects (p < 0.01). A shorter time since the latest booster vaccination was found to affect avidity positively on the next PT-antigen exposure with either vaccination or infection.

Bioethanol production from banana peels

Banana varieties have a significant effect on the reducing sugar content in the peel. In recent years, potential efforts have been directed towards the utilization of cheap renewable agricultural resources, such as banana peel waste as alternative substrate for ethanol production. Various conversion pathways are compared from technical, economic, and environmental points of view. This study also deals mainly with the yield of ethanol from molasses with respect to the dilution ratio and the amount of yeast used for fermentation keeping the temperature and fermentation duration constant. This fuel energy is also a safer substitute to methyl tertiary-butyl ether (MTBE), a common additive used in gasoline for clean combustion.

Determination of Nicotinic Acid in Food and Pharmaceuticals by a Simple and Rapid Fluorescence Polarization Immunoassay (FPIA)

A simple and rapid fluorescence polarization immunoassay (FPIA) based on polyclonal antibodies is reported for the first time for the determination of nicotinic acid. First, six nicotinic acid analogues were coupled with fluorescein isothiocyanate isomer I (FITC) to obtain tracers. The influences of tracer structure on the performance of FPIA as well as the structure-activity relationships were explored. Subsequently, the tracer concentration and antibody dilution ratio on the performance of FPIA were investigated. For the optimal conditions, the detection range of FPIA was from 1.1 to 145.6 μg mL−1 and its IC20 value as the limit of detection (LOD) was 1.1 μg mL−1. The cross-reactivity of antibodies with common vitamins was evaluated. The intra-assay recovery was from 92.0% to 101.2% in food and tablets, with inter-assay recoveries between 92.0% and 103.8%. The developed FPIA provided good sensitivity and reproducibility and therefore provided efficient and economic screening for nicotinic acid in food and tablets.

Critical Size and Formation Mechanism of Secondary Nuclei in Melt-Crystallized Polylactide Stereocomplex Crystals

Revealing the molecular mechanism and determining the length scale of the rate-limiting ordering process have been a challenge in the field of polymer crystallization. Previously, we proposed a theory to determine the size of critical nuclei in single-component crystals from the variation of nucleation kinetics with different dilution ratios. Here, we further extend this theory to polylactide (PLA) stereocomplex crystals (SCs) consisting of poly(l-lactide) and poly(d-lactide). By diluting only l-lactide units or both l-lactide and d-lactide units via random copolymerization, the number of l-lactide units and that of two types of units in a critical secondary nucleus were obtained. The results show that a critical secondary nucleus of an SC bulk crystal consists of 22–27 l-lactide units and the same number of d-lactide units when crystallized from melt at temperatures ranging from 140 to 170 °C. These repeating units form two poly(l-lactide) stems and two poly(d-lactide) stems. The four stems in a secondary nucleus of SC are chosen from approximately three to four chains. According to the number of chains, at most one-third of PLA chains has once adjacent re-entry folding in the secondary nuclei of SC crystallized from melt. These results provide rich information on the ordering process during the melt crystallization of SC crystals and are beneficial for understanding the crystallization mechanism.

Baseline and usual cardiorespiratory fitness and the risk of chronic kidney disease: A prospective study and meta-analysis of published observational cohort studies

Chronic kidney disease (CKD) is a global public health burden but is largely preventable. Few studies have reported an association between high levels of cardiorespiratory fitness (CRF) and decreased risk of CKD. However, some relevant aspects of the association, such as the dose-response relationship and the long-term relevance of CRF levels to CKD, have yet to be addressed. We, therefore, aimed to quantify the nature and magnitude of the association between CRF and CKD in a cohort of 2099 men aged 42-61 years with normal kidney function at baseline. CRF was directly measured using a respiratory gas exchange analyzer during cardiopulmonary exercise testing. Hazard ratios (HRs) (95% CIs) were estimated for the incidence of CKD. Correction for within-person variability was made using data from repeat measurements of CRF taken 11 years after baseline. A total of 197 CKD events were recorded during a median follow-up of 25.8 years. The age-adjusted regression dilution ratio for CRF was 0.59 (95% CI: 0.53-0.65). The risk of CKD decreased in a graded fashion with increasing CRF. Comparing extreme tertiles of CRF, the HR (95% CI) for CKD was 0.67 (0.46-0.97) following adjustment for several established and emerging risk factors. The corresponding adjusted HR (95% CI) was 0.51 (0.27-0.96) following correction for within-person variability. In a meta-analysis of five population-based cohort studies (including the current study) comprising 32,447 participants and 4,043 CKD cases, the fully-adjusted risk ratios (95% CIs) for CKD comparing extreme tertiles of baseline and long-term CRF values were 0.58 (0.46-0.73) and 0.40 (0.27-0.59), respectively. Findings from a new prospective study and pooled analysis of previous studies plus the new study indicate that high CRF levels are strongly and independently associated with a reduced risk of CKD and consistent with a linear dose-response relationship. Using single baseline measurements of CRF to investigate the association between CRF and CKD risk could considerably underestimate the true association.