Peak Height Ratio Research Articles

Exploring the combination of wilting and different types of additives to improve silage quality of highland alfalfa: Fermentation quality, nutritional values, molecular structural features, and ruminal degradability

The study aimed to explore the interaction between wilting pretreatment and different types of ensiling additives on silage quality, nutritional value, molecular structural features, and ruminal degradability of highland alfalfa. A total of 48 bags of alfalfa forage were randomly assigned into one of eight treatments in a 2 × 4 factorial arrangement (6 replication bags/treatment). One kg of either unwilted (72 % moisture) or wilted (60 %) alfalfa forage was inoculated with acidic salt, lactic acid bacteria, organic acids, or left without additive (control). Our results indicated that the wilting process increased lactic acid and crude protein levels, decreased the NH3-N level in silage, and increased its in vitro DM digestibility (all P < 0.05). The wilting treatment increased the amide I/II ratio by 31.7 % and the total carbohydrate II regions by 19.6 % (both P < 0.05). Inoculating pre-wilted alfalfa with lactic acid bacteria further reduced the pH and NH3-N levels and increased the lactic acid level and DM digestibility in silage (all P < 0.05). The addition of organic acids decreased silage pH and increased lactic acid, crude protein, and starch contents, and DM digestibility (all P < 0.05). Correlation analysis indicated that silage with a higher amide I/II peak height ratio and TC2 areas had more digestible nutrients and higher DM digestibility. In conclusion, wilting pretreatment combined with lactic acid bacteria inoculant is an effective approach to improve quality and nutrient availability of highland alfalfa silage. The non-invasion spectrometer has the potential to reveal the molecular spectral characteristics of alfalfa silage and predict nutrient availability.

Synthesis and EPR Studies of Zinc and Copper TPMA Metal Complexes Containing TEMPO Functionalities

A new family of stable radical‐containing polytopic ligands based on tris(2‐pyridylmethyl)amines TPMA architecture and their corresponding metal complexes have been synthesized. These molecular systems offered the possibility to investigate how various spin carriers, such as the metal ion and the organic radical, influence their EPR properties mainly through the empirical ratio of peak heights d1/d. Moreover, it has been possible to analyze how different conformations of TEMPO radical units in the molecular skeleton affect the metallic system.

High-precision analysis of carbon isotopic composition for individual CO2 inclusions via Raman spectroscopy: Addressing issues arising from the laser-heating effects

The carbon isotopic composition of carbon dioxide fluid inclusions trapped within minerals reflects the origin and evolution of the CO2-bearing fluids and melts. Recently, we developed a highly accurate method using Raman spectroscopy to determine the carbon isotopic composition of CO2 fluid inclusions, by simultaneously collecting 13CO2 and hot bands at a controlled temperature, and using the peak height ratio of 13CO2 and one of hot bands near 1410 cm−1 (HR) to determine δ13C value. This microanalytical technique enables precise measurements of the δ13C values of different CO2 fluid inclusions within zoned mineral crystals to reveal the multiple-stages evolution of CO2-bearing fluids and melts. However, when studying fluid inclusions hosted within high laser heating coefficient minerals such as olivine and pyroxene in mantle xenoliths, local temperature near the inclusions will be higher than the temperature controlled by heating-cooling stage. To address this issue, we examined the effects of laser heating on CO2 isotopic standard samples sealed in the silica capillary, and developed a novel procedure to determine the δ13C values of natural fluid inclusions by using two CO2 standard samples with known carbon isotopic compositions. The proposed workflow successfully determines the δ13C values of natural CO2 inclusions, with an average deviation of 0.58 ‰ between the measurements and the model.

Micro FT-IR spectroscopic measurements of CH4: Experimental calibration with high-pressure and high temperature optical cell (HPOC)

Fourier transform infrared spectroscopy (FTIR) is a prevalent nondestructive in situ analytical technique widely employed for the qualitative characterization of natural fluid inclusions. Presently, the quantitative determination of the temperature, pressure, and composition of natural inclusions stands as a pivotal challenge in the geological application of laser spectroscopy. Through the integration of the capillary high-pressure optical cell (HPOC) technique and Fourier transform infrared spectroscopy, infrared quantitative models for the C–H symmetric stretching band (v3) in the vapor phase of CH4 under varying temperature (40–200 °C) and pressure (20–500 bar) conditions are established for the first time. This accomplishment is achieved through fitting and calculation of infrared spectra. A linear quantitative relationship and the dynamic evolution of infrared spectral parameters (peak area, full width at half maximum, peak height ratio, and peak shifts) concerning temperature, pressure, and density are meticulously investigated. The findings reveal that the peak area and full width at half maximum exhibit an upward trend with increasing pressure and density, and are inversely proportional to temperature. The height ratio (P band/Q band, R band/Q band) experiences an increase as density escalates. Moreover, the CH4v3 band position shows a higher wavenumber shift with increasing temperature, while its position correlates negatively with pressure and density. Thus, this method demonstrates its applicability in deducing the PVT-x properties of CH4-bearing systems, including CH4-rich fluid inclusions, CH4 content within hydrocarbon inclusions, and experimental fluid investigations. In contrast to the Raman quantitative approach, the infrared quantitative formula can be universally adopted in any infrared laboratory owing to the quantitative theory (Beer–Lambert law) governing infrared absorption spectra. By collecting Raman spectra and infrared spectra of pure methane fluid inclusions from the central region of the Alps and subsequently calculating the results using two distinct quantitative methods for fluid inclusions, we have demonstrated the feasibility of applying infrared spectroscopy in natural fluid inclusion studies.

Developmental validation of a novel 8-dye fluorescent typing system with 59 Y-STRs and 3 Y-indels for forensic applications.

An 8-dye fluorescence-labeling forensic Y-chromosomal short tandem repeats (Y-STRs) kit, the 62-plex Y-STR multiplex amplification system, was developed and optimized. The system was validated by testing PCR conditions, stutter ratios (SR) and peak height ratios, sensitivity, mixture samples, precision and accuracy, species-specificity, and inhibition studies according to the Scientific Working Group on DNA Analysis Methods guidelines. PCR-based studies showed that the recommended PCR conditions were optimized for this kit. In the sensitivity study, a full profile was obtained from template DNA with a quantity of u125pg. Consistent profiles were obtained from three different laboratories. The SRs in all loci were less than 15%, and nice balance and suitable average peak height were shown. No peaks were detected in the profiles of common animal species and microorganisms. In the male-male mixture studies, all loci were observed at a ratio of 1:8, and in the male-female mixture study, all alleles could be profiled at a ratio of 1:500 if the male DNA inputs were ≥0.5ng/µL. An inhibitor study demonstrated that the kit had varying degrees of resistance to the presence of common inhibitors. Population study demonstrated the 62-plex Y-STR Kit improved the power of discrimination in unrelated Chinese Han males (n=192). When haplotype diversity was 1, the probability of discrimination power of the 62-plex Y-STR Kit was 0.9948, which is suitable for forensic investigations. The results show that the developed 8-dye fluorescence labeling 62 loci system is sensitive, robust, convenient, and highly informative for forensic applications.

Performance comparison of four qPCR and three autosomal STR commercial kits from degraded skeletal remains

This research evaluates the current DNA quantification (Quantifiler™ Trio, PowerQuant®, Investigator® Quantiplex® Pro and InnoQuant® HY Fast) and autosomal STRs amplification kits (GlobalFiler™, PowerPlex® Fusion 6 C, Investigator® 24Plex QS) using 62 degraded skeletal remains from armed conflicts (petrous bone, femur, tibia, and tooth) with several parameters (autosomal small, large, and male target, degradation index, probability of degradation, number of alleles above analytical threshold, number of alleles above stochastic threshold, RFU, peak height ratio, number of reportable loci). The best qPCR/autosomal STRs amplification tandem was determined by comparing quantification results by a DNA quantity estimation based on sample average RFU. InnoQuant® HY Fast was the most sensitive kit, and no significative differences were observed among amplification kits; however, Investigator® 24 Plex QS was found to be the most sensitive in our samples. That is why InnoQuant™ and Investigator® 24Plex QS were determined to be the best tandem.

Alternative direct-to-amplification cell lysis techniques for forensically relevant non-sperm cells.

While efforts have been made to reduce the pervasive backlog of sexual assault evidence collection kits, the actual laboratory process remains very time-consuming due to the requirement of a differential lysis step before DNA purification, as well as intricate mixture analysis towards the end of the DNA workflow. Recently, an alternative, direct-to-amplification sperm lysis method (using 1 M NaOH) was identified. However, a direct cell lysis method for non-sperm cells has not been identified yet. Thus, the primary objective of this work was to find an alternative method that is quick, inexpensive, and does not require multiple purification steps for the lysis of non-sperm cells in sexual assault samples. In this study, vaginal swab samples were lysed with the control method, prepGEM™, as well as six alternative reagents: alkaline buffer with 25-200 mM NaOH, high-salt stain extraction buffer, modified radioimmunoprecipitation assay (RIPA) buffer, mammalian protein extraction reagent (M-PER™), digitonin buffer, and urea/thiourea buffer. Quantification using Quantifiler® Trio of vaginal and semen lysates revealed that the alkaline (25 mM NaOH) and M-PER™ methods were efficient for the lysis of vaginal epithelial cells without substantial sperm cell lysis. Following quantification, analysis of STR profiles from vaginal lysates revealed that the M-PER™ method showed promising results across all metrics examined, including the percentage of detected STR alleles, mean peak heights, peak height ratio, and interlocus balance. Thus, this method was recommended as an alternative to the traditional differential lysis method for non-sperm cells given its ability to produce amplification-ready lysates without any DNA purification step.

Application of real-time PCR-based multicolor melting curve with automatic analysis system in pregestational and prenatal thalassemia diagnoses.

To evaluate the value of the real-time PCR-based multicolor melting curve analysis (MMCA) with an automatic analysis system used in a mass thalassemia screening and prenatal diagnosis program. A total of 18,912 peripheral blood samples from 9456 couples and 1150 prenatal samples were detected by MMCA assay. All prenatal samples were also tested by a conventional method. Samples with unknown melting peaks, unusual peak height ratios between a wild allele and a mutant allele, or a discordant phenotype-genotype match were further studied by using multiplex ligation-dependent probe amplification (MLPA) or Sanger sequencing. All MMCA results were automatically analyzed and manually checked. The consistency between MMCA assay and conventional methods among prenatal samples was investigated. Except for initiation codon (T>G) (HBB:c.2T>G), all genotypes of thalassemia inside the scope of conventional methods were detected by MMCA assay. Additionally, 27 carriers with 10 rare HBB variants, 13 with α fusion gene, 1 with a rare deletion in α globin gene, and 1 with rare HBA variant were detected by using MMCA assay. MMCA can be an alternative approach used in routine thalassemia carrier screening and prenatal diagnosis for its high throughput, sufficient stability, low cost, and easy operation.

Multiparametric, Label-Free Analysis of Microfluidic Droplets via a Dynamic Phase Grating Approach.

Droplet-based microfluidic systems possess many fundamental advantages as a platform for the analysis of chemical and biological species. However, whereas on-chip operations have rapidly developed over the past decades, approaches for analyzing target molecules within droplets have largely remained limited to methods requiring bulky and expensive instrumentation. In this work, we describe a droplet analysis approach whereby the droplet train itself is the sensing construct. Specifically, the droplet train is interrogated as a transmission phase grating, allowing high-throughput, label-free, solution-phase, and multi-parametric analysis of droplet contents. Importantly, three distinct properties of generated droplets can be simultaneously extracted using this conceptually simple and experimentally straightforward measurement approach. Under constant droplet generation conditions, measurement of droplet viscosity is achieved by monitoring changes in zero order to first order peak separation in the far-field diffraction pattern, with a sensitivity of 2.28 × 10-4 cSt per μm change in peak separation. In parallel, measurement of droplet refractive index (RI) is achieved by measuring changes in the ratio of the zero order to first order peak intensity, with a sensitivity of 2.14 × 10-4 RI units per unit change in a diffracted peak intensity ratio. Finally, droplet generation frequency is determined from the time-varying oscillation of the peak height ratio, yielding comparable results to an expensive high-speed camera commonly used for droplet imaging. Importantly, the experimental strategy for this approach is straightforward and does not require expensive instrumentation; therefore, it may find utility in affordable and portable analysis approaches applied to diverse droplet microfluidic assays.

Comparative analysis of two Rapid DNA technologies for the processing of blood and saliva-based samples

Rapid DNA technologies recently gained significant momentum as a means to generate DNA profiles faster than with standard laboratory workflows. Initially developed for the analysis of buccal reference samples, applications are being considered for other types of forensic samples. In this study, an identical set of 150 blood and saliva-based samples was processed using two different Rapid DNA technologies, the Applied BioSystems™ RapidHIT™ ID System using the RapidINTEL™ sample cartridge and the ANDE™ 6C Rapid DNA Analysis™ System using the I-Chip. A subset of samples were subjected to alternative collection methods or sample pre-treatments to determine the optimal strategy for each instrument. An equivalent sample set was also processed using a conventional DNA analysis workflow. The sensitivity range of the two Rapid DNA technologies was comparable based on blood and saliva dilution series, with both technologies able to generate full profiles from samples typically yielding 5–10 ng of DNA when processed using conventional DNA analysis. The brand of cotton swabs used for Rapid DNA analysis had an impact on the results for both systems. Differences were observed in success rate between the two systems when processing blood (on fabrics, FTA paper or hard surfaces) and saliva-based samples (drink containers, FTA paper, chewing gum, cigarette butt filter paper) and depended on the sample type. Importantly, deviating from the manufacturer’s instructions for sample collection and pre-treatment was more detrimental to the ANDE 6C results. The quality of DNA profiles, as assessed using heterozygote peak height ratios, interloci balance and artifact presence, confirmed the results to be reliable and acceptable for single source samples. Profiling results were obtained when samples were reprocessed using the same Rapid DNA technology or conventional DNA analysis. Secondary analysis using a substitute software (GeneMapper ID-X v1.5) to recover additional genetic information was shown to be feasible. Finally, a comparison between the Applied Biosystems™ RapidHIT™ ID System Software v1.3.1 and v1.3.2 was also performed. Findings of this study could assist those interested in using Rapid DNA technology for blood or saliva-based samples, in various settings and for different applications.

Development of a Quantitative Multiplex PCR to Detect Three Common Alpha Thalassemia Deletions

Alpha thalassemia is an autosomal recessive genetic disorder with a high prevalence in the Middle East. The severe form of alpha-thalassemia is incompatible with life and can cause significant obstetric complications in the mother. Therefore, it is important to determine the genotype in parents who have a chance of having a fetus with one of the severe forms of this disease. A total of 112 samples that were previously analyzed for common alpha thalassemia mutations in Iran were used in this study. A new multiplex PCR including quantitative polymerase chain reaction to amplify the homologous regions of the alpha-globin gene cluster and fluorescent gap PCR was designed to identify −α3.7, −α4.2, --MED deletions. The ROC curve was used to determine the optimum cutoff points. Statistical analysis showed that there is a significant difference between the peak height ratios for different genotypes. The peak corresponding to the 297 bp fragment resulting from the amplification of the allele with MED-I deletion was detected in all the samples with this deletion. Different cutoffs for a range of sensitivities and specificities were determined by the ROC curve. The suggested method can identify three common large deletions in the alpha-globin gene cluster. A study with a larger sample size can provide more accurate information about the sensitivity and specificity of this test.

Effects of an Acidic Environment on the Strength and Chemical Changes of Resin-based Composites.

The objective of this study was to evaluate the aging effects of long-term exposure to acidic beverages on the flexural strength (FS) and chemical reactions of two resin-based composites (RBCs) and one giomer. The FS of composite specimen bars (2 mm × 2 mm × 25 mm) was measured using a universal testing machine at various levels of thermocycling (TC; 0, 10,000, 50,000, and 100,000 cycles) in two beverages with different pH values (distilled water [DW], pH 7.0; Coca-Cola, pH 2.4-2.8). The FS data were analyzed using three-way analysis of variance with the post hoc Tukey test and t-test at a significance level of a=0.05. In DW, the FS of an RBC and a giomer did not decrease until 10,000 cycles. The other RBC, Z250, decreased rapidly until 50,000 cycles (p<0.05), followed by no additional decrease until 100,000 cycles. In Coca-Cola, the FS of two RBCs and a giomer decreased more rapidly than in DW from 10,000 cycles (t-test, p<0.05). In Coca-Cola, the increased porosity observed in scanning electron microscopy (SEM) images, the changes of the hydroxyl peak at 3340 cm-1 and ester peak at 1730-1700 cm-1 in Fourier-transform infrared spectroscopy in attenuated total reflectance mode (FTIR-ATR) spectra, and the continuous increase of the Si-O/Si-C peak height ratio from 10,000 cycles to 100,000 cycles in X-ray photoelectron spectroscopy (XPS) suggested an increased loss of silane-carbon bond between the matrix and fillers of the Z250 RBC, compared to those in DW. In conclusion, when TC was performed in DW, unreacted monomers and a coupling agent were washed out, which caused porosity and reduced FS. In Coca-Cola, acidic conditions accelerated the removal of the matrix through the hydrolysis reaction at the ester groups, resulting in more porosity and a faster decrease in FS than in DW.

Predicting TERT promoter mutation status using 1H-MR spectroscopy and stretched-exponential model of diffusion-weighted imaging in IDH-wildtype diffuse astrocytic glioma without intense enhancement.

Isocitrate dehydrogenase (IDH)-wildtype diffuse astrocytic glioma with telomerase reverse transcriptase (TERT) promoter mutation is defined as glioblastoma by the WHO 2021 criteria, revealing that TERT promotor mutation is highly associated with tumor aggressiveness. The aim of this study was to identify features from MR spectroscopy (MRS) and multi-exponential models of DWI distinguishing wild-type TERT (TERTw) from TERT promoter mutation (TERTm) in IDH-wildtype diffuse astrocytic glioma. Participants comprised 25 adult patients with IDH-wildtype diffuse astrocytic glioma. Participants were classified into TERTw and TERTm groups. Point-resolved spectroscopy sequences were used for MRS data acquisition. DWI was performed with 13 different b-factors. Peak height ratios of NAA/Cr and Cho/Cr were calculated from MRS data. Mean apparent diffusion coefficient (ADC), perfusion fraction (f), diffusion coefficient (D), pseudo-diffusion coefficient (D*), distributed diffusion coefficient (DDC), and heterogeneity index (α) were obtained using multi-exponential models from DWI data. Each parameter was compared between TERTw and TERTm using the Mann-Whitney U test. Correlations between parameters derived from MRS and DWI were also evaluated. NAA/Cr and Cho/Cr were both higher for TERTw than for TERTm. The α of TERTw was smaller than that of TERTm, while the f of TERTw was higher than that of TERTm. NAA/Cr correlated negatively with α, but not with other DWI parameters. Cho/Cr did not show significant correlations with any DWI parameters. The combination of NAA/Cr and α may have merit in clinical situation to predict the TERT mutation status of IDH-wildtype diffuse astrocytic glioma without intense enhancement.

Rapid detection of the biomarker for cystinuria by a metal-organic framework fluorescent sensor

Arginine (Arg) is considered a valuable biomarker for various diseases, including cystinuria, and its concentration level holds significant implications for human health. To achieve the purposes of food evaluation and clinical diagnosis, it is imperative to develop a rapid and facile method for selective and sensitive determination of Arg. In this work, a novel fluorescent material (Ag/Eu/CDs@UiO-66) was synthesized by encapsulating carbon dots (CDs), Eu3+ and Ag + into UiO-66. This material can serve as a ratiometric fluorescent probe for detecting Arg. It exhibits a high sensitivity with a detection limit of 0.74 μM and a relatively broad linear range from 0–300 μM. After dispersing the composite Ag/Eu/CDs@UiO-66 in an Arg solution, the red emission of Eu3+ center at 613 nm was significantly enhanced, while the characteristic peak of CDs center at 440 nm remained unchanged. Therefore, a ratio fluorescence probe could be constructed based on the peak height ratio of the two emission peaks to achieve selective detection of Arg. In addition, the remarkable ratiometric luminescence response induced by Arg results in a significant color transition from blue to red under UV-lamp for Ag/Eu/CDs@UiO-66, which was convenient for visual analysis.

RP-HPLC Method Developed for the Determination of Metformin in Human Saliva

Several extraction steps involved while analyzing metformin in plasma necessitates the need to develop a simple less tedious RP-HPLC method for metformin analysis in human saliva.Blank saliva (2 mL) was spiked with 2 mL solution (12.5 µg mL-1) of metformin and 1 mL solution (0.5 µg mL-1) of caffeine as internal standard (IS). The mixture was vortex mixed and centrifuged at 3000 rpm for 10 minutes. A portion (0.5 mL) of the resultant solution was injected into the HPLC machine (Agilent 1260 infinity). The optimized conditions included a mobile phase of methanol:water (80:20 v/v) containing 0.1 % orthophosphoric acid, isocratic elusion mode, an injection volume of 10 µL, flow rate of 1 mLmin-1, at 35°C and detection wavelength of 232nm. Calibration curve(1.25to25.0 µg mL-1) was prepared by plotting the peak height ratios of metformin Vs IS against their corresponding concentrations. The method was validated according to ICH guidelines. Metformin and caffeine eluted at 1.6 and 2.6 minutes respectively. The method was precise (&lt;1 % RSD), accurate (% Er of 1.00 and % recovery of 99.98 %) with linear calibration curve (r= 0.9987). The developed method can be used for determination of metformin in human saliva. Keywords: Metformin, Saliva, Isocratic elution, RP-HPLC

Ratiometric fluorescent sensor based on metal–organic framework for selective and sensitive detection of CO32–

Carbonate ion (CO32–) is an anion essential for the maintenance of life activities and is of great importance to human health. Here, a novel ratiometric fluorescent probe Eu/CDs@UiO-66-(COOH)2 (ECU) was prepared by introducing europium ions (Eu3+) and carbon dots (CDs) into the UiO-66-(COOH)2 parent framework under the guidance of a post-synthetic modification strategy and used for the detection of CO32– ion in the aqueous environment. Interestingly, when CO32– ions were added to the ECU suspension, the characteristic emission of carbon dots at 439 nm was significantly enhanced, while the characteristic emission of Eu3+ ions at 613 nm was reduced. Therefore, the detection of CO32– ions can be realized through the peak height ratio of the two emissions. The probe had a low detection limit (about 1.08 μM) and a wide linear range (0–350 μM) for the detection of carbonate. In addition, the presence of CO32– ions can cause a significant ratiometric luminescence response and resulted obvious red-to-blue color shift of the ECU under UV light, which will facilitate visual analysis by the naked eye.

Effects of selected solvents on PMMA after prolonged exposure: unilateral NMR and ATR-FTIR investigations

Exposure of heritage plastics to solvents can cause several changes to their surface layers and bulk. The amorphous nature of certain plastics, particularly polymethyl methacrylate (PMMA), allows penetration and diffusion of low-molecular liquids, which can lead to swelling, cracking, plasticisation or stiffening, extraction and dissolution of polymer and additive components. Such phenomena compromise the visual characteristics, chemical and physical structure of heritage collections and remain, for the most part, unexplored. Research studies present contradicting evidence concerning the use of solvents and their safe application on PMMA. The main discrepancy is about PMMA being permeable to water or not. Alcohols have also triggered debates about whether they cause leaching and/or depolymerisation. This paper investigates the potential effects that selected free polar and non-polar solvents—employed in different media as cleaning systems—can have on PMMA after prolonged contact. The Hildebrand solubility theory was used to select solvents based on PMMA’s miscibility. The effects of deionised water, ethanol, isopropanol and petroleum ether are examined through a 30-day immersion study of PMMA via microscopic examination, weight change measurements, ATR-FTIR, peak height ratios, and NMR MOUSE. All four solvents extracted soluble components, i.e. unreacted residual monomer, and increased PMMA’s surface sensitivity to abrasions. Water and petroleum ether did not cause any measurable chemical or physical changes. Alcohols caused surface crystallisation and crazing, with isopropanol leading to stiffening and ethanol to plasticisation of the material. These effects, although extreme and absent in most routine cleaning timeframes, demonstrate the cumulative damage these solvents can potentially induce to PMMA.

Quantitative Analysis of Silicon Tetrachloride, Carbon Disulfide, and Dichloroethane Concentration by Raman Spectroscopy

Quantitative analysis of silicon tetrachloride, carbon disulfide, and dichloroethane concentrations to obtain vapor-liquid equilibrium data of the SiCl4-CS2 and SiCl4-C2H4Cl2 binary systems was established by Raman spectroscopy. The cheap glass sampling pipe was used as a carrier for Raman spectroscopy measurements. The Raman peak height of the internal standard was used to remove interference factors such as sampling pipe diameter, temperature, laser power, and other effects from the instrument. The peak height ratio between the Raman characteristic peak of the analyte and that of the internal standard was proportional to the analyte concentration. During the measuring process of vapor-liquid equilibrium data for the SiCl4-C2H4Cl2 binary system, the linear equation of y = 0.0068 + 0.75x with R2 of 0.9939 was used for the determination of SiCl4 concentration at the 422 cm−1 band. The linear equation of y = 0.0019 + 0.2266x with R2 of 0.9966 was used for the determination of C2H4Cl2 concentration at the 754 cm−1 band. For the SiCl4-CS2 binary system, the linear equation of y = 0.0494 + 4.7535x with R2 of 0.9962 was used for the determination of SiCl4 concentration at the 422 cm−1 band. The linear equation of y = 0.8139 + 8.7366x with R2 of 0.9973 was used for the determination of CS2 concentration at the 654 cm−1 band. The concentration of standard samples calculated by these standard curves was compared with the actual value to verify the accuracy of this method. The reproducibility is good when determining silicon tetrachloride and dichloroethane concentrations for the SiCl4-C2H4Cl2 binary system, with RSEP values of 2.81% and 2.17%, respectively. Meanwhile, the RSEP values are 3.55% and 4.16%, respectively, when determining silicon tetrachloride and carbon disulfide concentrations for the SiCl4-CS2 binary system.

Target-allele-specific probe single-base extension (TASP-SBE): a novel MALDI-TOF-MS strategy for multi-variants analysis and its application in simultaneous detection of α-/β-thalassemia mutations.

Single-nucleotide variants (SNVs) and copy number variations (CNVs) are the most common genomic variations that cause phenotypic diversity and genetic disorders. MALDI-TOF-MS is a rapid and cost-effective technique for multi-variant genotyping, but it is challenging to efficiently detect CNVs and clustered SNVs, especially to simultaneously detect CNVs and SNVs in one reaction. Herein, a novel strategy termed Target-Allele-Specific Probe Single-Base Extension (TASP-SBE) was devised to efficiently detect CNVs and clustered SNVs with MALDI-TOF-MS. By comprehensive use of traditional SBE and TASP-SBE strategies, a MALDI-TOF-MS assay was also developed to simultaneously detect 28 α-/β-thalassemia mutations in a single reaction system, including 4 α-thalassemia deletions, 3 HBA and 21 HBB SNVs. The results showed that all 28 mutations were sensitively identified, and the CNVs of HBA/HBB genes were also accurately analyzed based on the ratio of peak height (RPH) between the target allele and reference gene. The double-blind evaluation results of 989 thalassemia carrier samples showed a 100% concordance of this assay with other methods. In conclusion,a one-tube MALDI-TOF-MS assay was developed to simultaneously genotype 28 thalassemia mutations. This novel TASP-SBE was also verified a practicable strategy for the detection of CNVs and clustered SNVs, providing a feasible approach for multi-variants analysis with MALDI-TOF-MS technique.

Classification and surgical treatment methods for partial traumatic upper lip deformity.

Establish a classification method for partial traumatic upper lip deformity and verify the classified repair method is superior to the traditional non-classified method. Lip deformities caused by partial tissue defects of less than one-third of total lip length were classified into three types and conducted corresponding surgery based on philtrum ridge and orbicularis oris muscle involvement as well as the extent of roll line discontinuity. In this review, 42 cases were non-classified historical controls before the classification was devised, and 67 cases were classified. Data were collected 12months after surgery. The pre- and post-operative data of all patients were compared. In classified patients, the scar width decreased significantly, from 3.1± 0.6 mm to 1.2± 0.2 mm; the height difference of the groove line was significantly reduced from 3.3± 0.9 mm to 0.9± 0.1 mm; the ratio of the vermilion area of the affected to healthy side decreased significantly from 1.37 ± 0.31 to 1.05 ± 0.17; the ratio of the lip peak height of the affected to healthy side in type III decreased significantly from 1.91 ± 0.32 to 1.07 ± 0.12; patient satisfaction rate was about 98.5 percent. Clinical outcomes showed significant improvement of lip aesthetics with a high patient satisfaction rate in the classified group than the non-classified group.