Ultrafast quantitation of six quinolones in water samples by second-order capillary electrophoresis data modeling with multivariate curve resolution–alternating least squares

This paper presents the development of a capillary electrophoresis method with diode array detector coupled to multivariate curve resolution–alternating least squares (MCR-ALS) to conduct the resolution and quantitation of a mixture of six quinolones in the presence of several unexpected components. Overlapping of time profiles between analytes and water matrix interferences were mathematically solved by data modeling with the well-known MCR-ALS algorithm. With the aim of overcoming the drawback originated by two compounds with similar spectra, a special strategy was implemented to model the complete electropherogram instead of dividing the data in the region as usually performed in previous works. The method was first applied to quantitate analytes in standard mixtures which were randomly prepared in ultrapure water. Then, tap water samples spiked with several interferences were analyzed. Recoveries between 76.7 and 125 % and limits of detection between 5 and 18 μg L^?1 were achieved.     

Fourier Transform Infrared Spectroscopy of Skin Cancer Cells and Tissues

Abstract

The aim of this review is to combine all the information related to the characterization of skin tissues and cells, focusing on the identification of the specific biochemical characteristics of skin cancer. We have characterized two types of melanoma by FTIR spectroscopy: a murine melanoma (B16F10 cell line) and a human melanoma (C8161 cell line). The cells were deposited on IR transparent CaF₂ windows, the spectral range used lay between 900-4000 cm^{? 1}, transmission mode with 2 cm^{? 1} resolution, and 32 scans. A biochemical association is presented for all the absorption bands identified in this study. Besides the characterization of both cell lines above, a collection of works done in the area of skin cancer was also carried out, in this review; some interesting results obtained by different authors with respect to the characterization of different samples (melanomas, follicle sheath, basal cell carcinoma, epidermis tissues, dermis tissue, and human stratum corneum) are presented and compared to the chemical and biological associations performed in each case. The identification of biochemical injuries provides important information that, associated with clinical examination, can assist the diagnosis of diseases. Several FTIR techniques can be used in the diagnosis of biochemical changes in biological tissues, by identifying molecular markers associated with malignant and benign changes or variations in the composition of amino acids in tissues and cells. In the near future, a further study to compare histopatological analysis and biochemical characterization by FTIR spectroscopy would be interesting, in order to verify the accuracy and validate the applicability of this technique in the diagnosis of skin lesions from a statistical viewpoint.     

Speckle interferometry, fibre optic sensors and laser induced ultrasounds as solutions to industrial demands

Industrial operators have been taking interest in optical diagnostics through years. Optical methods are mainly well appreciated for their attitude to be used as non-contact and non-invasive techniques. The paper presents three examples of applications carried-out by researchers of Centro Elettrotecnico Sperimentale Italiano (CESI) in collaboration with people working in industrial fields. The first part shows the applications of speckle interferometry to measure residual stresses in association with the traditional blind-hole drilling, while the second part presents the installation of fibre optic sensors in a power plant for monitoring possible overheating to avoid fires and finally the last presentation outlines a particular application in the field of laser generated ultrasounds that is the monitoring of the variation of ultrasonic speed propagation due to residual stresses.     

Dispersive liquid–liquid microextraction of quinolones in porcine blood: Optimization of extraction procedure and CE separation using experimental design

A dispersive liquid–liquid microextraction procedure was developed to extract nine fluoroquinolones in porcine blood, six of which were quantified using a univariate calibration method. Extraction parameters including type and volume of extraction and dispersive solvent and pH, were optimized using a full factorial and a central composite designs. The optimum extraction parameters were a mixture of 250 μL dichloromethane (extract solvent) and 1250 μL ACN (dispersive solvent) in 500 μL of porcine blood reached to pH 6.80. After shaking and centrifugation, the upper phase was transferred in a glass tube and evaporated under N₂ steam. The residue was resuspended into 50 μL of water–ACN (70:30, v/v) and determined by CE method with DAD, under optimum separation conditions. Consequently, a tenfold enrichment factor can potentially be reached with the pretreatment, taking into account the relationship between initial sample volume and final extract volume. Optimum separation conditions were as follows: BGE solution containing equal amounts of sodium borate (Na₂B₄O₇) and di‐sodium hydrogen phosphate (Na₂HPO₄) with a final concentration of 23 mmol/L containing 0.2% of poly (diallyldimethylammonium chloride) and adjusted to pH 7.80. Separation was performed applying a negative potential of 25 kV, the cartridge was maintained at 25.0°C and the electropherograms were recorded at 275 nm during 4 min. The hydrodynamic injection was performed in the cathode by applying a pressure of 50 mbar for 10 s.     

Influence of ultrasonic waves in the reduction of nitrate to nitrite by hydrazine-Cu(II)

Colorimetric methods are still important for determining nitrate and nitrite. A critical step in the use of these methods to determine nitrate in low concentrations is the reaction time required to totally reduce nitrate to nitrite, i.e., 24h in the dark. This work involved a study of the influence of ultrasonic irradiation on the nitrate reduction reaction by hydrazine. Our findings indicated that ultrasonic irradiation, associated with copper(II) ion as a catalyst, increased the redox reaction rate, decreasing the reaction time to about 10min when the power of the ultrasonic irradiation was set in 14.0357W. The strong influence of the ultrasonic irradiation in the reduction reaction rates can be sustained by an excellent linear correlation (R(2)=0.9993) between the kinetic constants and ultrasonic powers. Nitrate conversion also increased from 68% to 98% at the latter conditions. It thus become clear that high intensity ultrasound is very beneficial for this reduction reaction to proceed in good yield and in short reaction time in comparison to its silent reaction.     

Hyperbranched rolling circle amplification as a rapid and sensitive method for species identification within the Cryptococcus species complex

The Cryptococcus species complex contains two closely related basidiomycetous yeasts: Cryptococcus neoformans and C. gattii, which cause cryptococcosis in humans and other animals. The species and varieties are characterized, by different clinical, epidemiological, biochemical and molecular features. The currently used identification methods are either time-consuming or not anymore commercially available. However, a rapid, sensitive and robust assay for the detection of these pathogens is vital for early diagnosis and appropriate treatment decisions. To overcome those limitations, four padlock probes targeting species-specific single nucleotide polymorphisms at the internal transcribed spacers (ITSs) of the RNA gene locus were developed and applied during isothermal hyperbranched rolling circle amplification (HRCA). The probes were tested against 99 samples, including 94 clinical cryptococcal cultures, three closely related Cryptococcus species, and two clinical specimens. The use of the padlock probes and the combination of probe signal amplification by HRCA provided a quick and sensitive assay for the accurate identification of C. neoformans var. grubii, C. neoformans var. neoformans and C. gattii. HRCA was also useful to detect hybrids, when they were heterozygous at the ITS locus. The HRCA results were in agreement with previous genotyping data based on PCR fingerprinting, amplified fragment length polymorphism and ITS sequencing.     

Selection and Optimization of Bacillus atrophaeus Inoculum Medium and its Effect on Spore Yield and Thermal Resistance

Bacillus atrophaeus's spores are used as biological indicators to monitor sterilization processes and as a Bacillus anthracis surrogate in the development and validation of biosafety methods. The regular use of biological indicators to evaluate the efficiency of sterilization processes is a legal requirement for health services. However, its high cost hinders its widespread use. Aiming at developing a cost-effective inoculum medium, soybean molasses and nutrient-supplemented vinasse were evaluated for their effectiveness in solid-state fermentation (SSF). In biomass production, the results demonstrated that all tested compositions favor growth by providing the nutritional demands of the microorganism. Optimum casein peptone and soybean molasses concentration (1.0%, 2.5%, or 4.0%) was determined by a 2((2-0)) factorial experimental design. The results have showed a positive influence of peptone on biomass production. In order to define peptone final concentration (4.0% or 6.0%), a 2(2) factorial experimental design was used. An optimized medium containing 4.0% soybean molasses and 4.0% casein peptone was similar in performance to a synthetic control medium (tryptone soy broth) in dry-heat thermal-resistant spore production by SSF. An experiment performed under optimum SSF conditions resulted in 1.9 x 10(10) CFU g(-1) dry matter with D (160 degrees C) = 5.2 +/- 0.2 min.     

Synthesis and structural study of a new series of 2‐methylsulfanyl‐tetrahydropyrimidines from β‐alkoxyvinyl trihalomethyl ketones

This work describes the synthesis of a novel series of 2‐methylsulfanyl‐tetrahydropyrimidines, from the cyclocondensation reaction of β‐alkoxyvinyl trihalomethyl ketones with 2‐methyl‐2‐thiopseudourea sulfate, in good yields. A detailed ¹H‐ and ¹³C‐NMR study was performed on the 2‐methylsulfanyl‐tetrahydropyrimidines obtained and 3D structures were proposed based on AM1 calculations supported by ¹H NMR coupling constants and NOESY experiments.