Adriana Barrinha Fernandes

Correlating the amount of urea, creatinine, and glucose in urine from patients with diabetes mellitus and hypertension with the risk of developing renal lesions by means of Raman spectroscopy and principal component analysis

Abstract. Patients with diabetes mellitus and hypertension (HT) diseases are predisposed to kidney diseases. The objective of this study was to identify potential biomarkers in the urine of diabetic and hypertensive patients through Raman spectroscopy in order to predict the evolution to complications and kidney failure. Urine samples were collected from control subjects (CTR) and patients with diabetes and HT with no complications (lower risk, LR), high degree of complications (higher risk, HR), and doing blood dialysis (DI). Urine samples were stored frozen (−20°C) before spectral analysis. Raman spectra were obtained using a dispersive spectrometer (830-nm, 300-mW power, and 20-s accumulation). Spectra were then submitted to principal component analysis (PCA) followed by discriminant analysis. The first PCA loading vectors revealed spectral features of urea, creatinine, and glucose. It has been found that the amounts of urea and creatinine decreased as disease evoluted from CTR to LR/HR and DI (PC1, p<0.05), and the amount of glucose increased in the urine of LR/HR compared to CTR (PC3, p<0.05). The discriminating model showed better overall classification rate of 70%. These results could lead to diagnostic information of possible complications and a better disease prognosis.

Quantifying creatinine and urea in human urine through Raman spectroscopy aiming at diagnosis of kidney disease

Abstract. Due to their importance in the regulation of metabolites, the kidneys need continuous monitoring to check for correct functioning, mainly by urea and creatinine urinalysis. This study aimed to develop a model to estimate the concentrations of urea and creatinine in urine by means of Raman spectroscopy (RS) that could be used to diagnose kidney disease. Midstream urine samples were obtained from 54 volunteers with no kidney complaints. Samples were subjected to a standard colorimetric assay of urea and creatinine and submitted to spectroscopic analysis by means of a dispersive Raman spectrometer (830 nm, 350 mW, 30 s). The Raman spectra of urine showed peaks related mainly to urea and creatinine. Partial least squares models were developed using selected Raman bands related to urea and creatinine and the biochemical concentrations in urine measured by the colorimetric method, resulting in r=0.90 and 0.91 for urea and creatinine, respectively, with root mean square error of cross-validation (RMSEcv) of 312 and 25.2  mg/dL, respectively. RS may become a technique for rapid urinalysis, with concentration errors suitable for population screening aimed at the prevention of renal diseases.

Detecting alterations of glucose and lipid components in human serum by near-infrared Raman spectroscopy

Introduction Raman spectroscopy may become a tool for the analysis of glucose and triglycerides in human serum in real time. This study aimed to detect spectral differences in lipid and glucose components of human serum, thus evaluating the feasibility of Raman spectroscopy for diagnostic purposes. Methods A total of 44 samples of blood serum were collected from volunteers and submitted for clinical blood biochemical analysis. The concentrations of glucose, cholesterol, triglycerides, and low-density and high-density lipoproteins (LDL and HDL) were obtained using standard biochemical assays. Serum samples were placed in Eppendorf tubes (200 µL), kept cooled (5 °C) and analyzed with near-infrared Raman spectroscopy (830 nm, 250 mW, 50 s accumulation). The mean spectra of serum with normal or altered concentrations of each parameter were compared to determine which Raman bands were related to the differences between these two groups. Results Differences in peak intensities of altered sera compared to normal ones depended on the parameter under analysis: for glucose, peaks were related to glucose; for lipid compounds the main changes occurred in the peaks related to cholesterol, lipids (mainly triolein) and proteins. Principal Components Analysis discriminated altered glucose, cholesterol and triglycerides from the normal serum based on the differences in the concentration of these compounds. Conclusion Differences in the peak intensities of selected Raman bands could be seen in normal and altered blood serum samples, and may be employed as a means of diagnosis in clinical analysis.

Effect of Ozone as Acaricide: Action of the Ozone on the Cuticle and Respiratory Spiracle of Tick Rhipicephalus sanguineus sensu lato

ABSTRACT Parasitoses that affect domestic animals, the Rhipicephalus sanguineus sensu lato deserves attention for being cosmopolitan. Ozone gas has been used as an agent for insect control. The objective of this study was to verify the acaricidic effect of the ozone gas on the R. sanguineus s. l. Ticks were distributed in three groups: two controls and one submitted to ozone action. Images were obtained using a scanning electron microscope. All specimens from the group treated with ozone died after the gas application and presented other lesions on cuticle and respiratory spiracles. Ozone acted as an acaricide to the ticks.

Raman spectroscopy applied to identify metabolites in urine of physically active subjects

Raman spectroscopy is a rapid and non-destructive technique suitable for biological fluids analysis. In this work, dispersive Raman spectroscopy has been employed as a rapid and nondestructive technique to detect the metabolites in urine of physically active subjects before and after vigorous 30min pedaling or running compared to sedentary subjects. For so, urine samples from 9 subjects were obtained before and immediately after physical activities and submitted to Raman spectroscopy (830nm excitation, 250mW laser power, 20s integration time) and compared to urine from 5 sedentary subjects. The Raman spectra of urine from sedentary showed peaks related to urea, creatinine, ketone bodies, phosphate and other nitrogenous compounds. These metabolic biomarkers presented peaks with different intensities in the urine of physically active individuals after exercises compared to before, measured by the intensity of selected peaks the Raman spectra, which means different concentrations after training. These peaks presented different intensity values for each subject before physical activity, also behaving differently compared to the post-training: some subjects presented increase while others decrease the intensity. Raman spectroscopy may allow the development of a rapid and non-destructive test for metabolic evaluation of the physical training in active and trained subjects using urine samples, allowing nutrition adjustment with the sports performance.

Identification and quantification of β-caryophyllene in copaiba oil using Raman spectroscopy

ABSTRACT Copaiba oleoresin presents several compounds with known biologic activity and physiologic effects, including analgesic and insecticide properties. Among them are the terpenoids (mainly diterpenes and sesquiterpenes) with β-caryophyllene, the main representative of the terpenoids and considered to be a chemical marker. This study employed Raman spectroscopy and principal component analysis (PCA) techniques to identify and quantify the β-caryophyllene marker in copaiba oil samples purchased from popular markets in Brazil. A dispersive Raman spectrometer (830 nm, 250 mW, 2 cm−1 spectral resolution) was used. Results showed the identification of the main Raman peaks from the β-caryophyllene in copaiba oil samples (main peaks at 507, 771, 1442, 1638, and 1673 cm−1). The loading vector 2 (PC2) extracted the spectral information from β-caryophyllene in the samples and the eigenvalue 2 (score 2) allowed the estimation of the concentration of this marker in commercial samples, with the concentrations from 15 to 34%. Raman spectroscopy combined with PCA may be considered to be a potential analytical tool for the quality control of Copaifera oil samples by quantifying β-caryophyllene using its unique spectral information.

Determining the amounts of urea and glucose in urine of patients with renal complications from diabetes mellitus and hypertension by near-infrared Raman spectroscopy

Diabetes mellitus and hypertension diseases are frequently found in the same patient, which if untreated predispose to atherosclerotic and kidney diseases. The objective of this study was to identify potential biomarkers in the urine of diabetic and hypertensive patients through dispersive near-infrared Raman spectroscopy. Urine samples were collected from patients with diabetes and hypertension but no complications (LG), high degree of complications (HG), and control ones: one fraction was submitted to biochemical tests and another one was stored frozen (-20°C) until spectral analysis. Samples were warmed up and placed in an aluminum sample holder for Raman spectra collection using a dispersive spectrometer (830 nm wavelength, 300 mW laser power and 20 s exposure time). Spectra were then submitted to Principal Components Analysis. The PCA loading vectors 1 and 3 revealed spectral features of urea/creatinine and glucose, respectively; the PCA scores showed that patients with diabetes/hypertension (LG and HG) had higher amount of glucose in the urine compared to the normal group (p < 0.05), which can bring serious consequences to patients. Also, the PCA scores showed that the amount of urea decreased in the groups with diabetes/hypertension (p < 0.05), which generates the same concern as it is a marker that has a strong importance in the metabolic changes induced by such diseases. These results, applied to the analysis of urine of patients with diabetes/hypertension, can lead to early diagnostic information of complications and a possible disease prognosis in the patients where no complications from diabetes and hypertension were found.

Use of photodynamic therapy in the treatment of bovine subclinical mastitis

BACKGROUND Bovine mastitis is a disease that causes a severe drawback in dairy production. Conventional treatments with antibiotic could leave antibiotic residues in the milk. The aim of this study was to evaluate the effect of photodynamic therapy in the treatment of bovine subclinical mastitis to develop an in vivo therapeutic protocol that could be used in routine farm practice, favoring the early return to production. METHODS Forty cows with subclinical mastitis (n = 40) were divided into 4 groups (control, photodynamic therapy - PDT, light irradiation - LED, and photosensitizer - PS). Control group received no treatment, PDT group received application of 1.0 mL of 2.5% toluidine blue photosensitizer followed by LED irradiation at λ = 635 nm, the LED group was treated with LED irradiation alone, and the PS group received only 2.5% toluidine blue dye. LED irradiation was applied to the mammary gland by means of an acrylic light guide coupled to the LED equipment. The PDT and LED groups were irradiated with 200 J/cm2 at three different positions inside the mammary gland. Milk samples were collected at 0 h, 12 h, 24 h after treatment for microbial identification and total bacterial count. RESULTS The treatment of the PDT group showed significant difference p < 0.05, characterizing the efficiency of this technique with the reduction of the microorganisms Streptococcus dysgalactiae and coagulase-negative Staphylococcus. CONCLUSION Photodynamic therapy was effective when applied in vivo for subclinical bovine mastitis. There was no need to separate the animal from production.

Effect of Ozone on Engorged Rhipicephalus microplus (Acari: Ixodidae) Females During the Pre-Laying Period

ABSTRACT The Rhipicephalus microplus is an important ectoparasite of livestock as it results in loss to the producer. The present study analyzed the effects of ozone gas, a fluid with acaricidal potential, on this species of cattle tick. Engorged females of R. microplus were divided into two groups: control, oxygen and ozone groups. The results indicated that ticks in the control and oxygen groups maintained the oviposition process intact. Females exposed to the ozone gas died before eggs were deposited and damage on the cuticle was observed, proving that the ozone gas had an acaricidal effect on the ticks.

Analysis of Damage on the Streptococcus mutans Immersed in Ozonated Water: Preliminary Study for Application as Mouth Rinse

ABSTRACT Ozonated water has been demonstrated to induce significant results in terms of the elimination of microorganisms. The present study assessed the damage to Streptococcus mutans after exposure to ozonated water; the ozone generator was adjusted to provide an outlet concentration of 60 mg/L, the samples were submitted to different ozonation times 1, 2, 4, 6, and 10 mi. Scanning electron microscopy and atomic force images were obtained to identify damage to the bacteria, followed by reactive oxygen species (ROS) evaluation and microbial viability. The results showed a significant reduction in viability and the images evidenced the generation of gaps on the microbial wall and surface layer alterations. Ozone can induce significant damage to S. mutans, thus suggesting that the use of ozonated water to prevent carious lesion formation is extremely promising.