Toxoplasmosis is an important zoonosis in public health because domestic cats are the main agents responsible for the transmission of this disease in Brazil. We investigate a method for diagnosing toxoplasmosis based on Raman spectroscopy. Dispersive near-infrared Raman spectra are used to quantify anti-Toxoplasma gondii (IgG) antibodies in blood sera from domestic cats. An 830-nm laser is used for sample excitation, and a dispersive spectrometer is used to detect the Raman scattering. A serological test is performed in all serum samples by the enzyme-linked immunosorbent assay (ELISA) for validation. Raman spectra are taken from 59 blood serum samples and a quantification model is implemented based on partial least squares (PLS) to quantify the sample's serology by Raman spectra compared to the results provided by the ELISA test. Based on the serological values provided by the Raman/PLS model, diagnostic parameters such as sensitivity, specificity, accuracy, positive prediction values, and negative prediction values are calculated to discriminate negative from positive samples, obtaining 100, 80, 90, 83.3, and 100%, respectively. Raman spectroscopy, associated with the PLS, is promising as a serological assay for toxoplasmosis, enabling fast and sensitive diagnosis.
Biomedical applications of near-infrared Raman spectroscopy have increased their importance at the last ten
years. This technique can determinate the molecular composition of materials, allowing a sensible and fast
biological diagnosis. It has showed to be a promising tool for health diagnosis due to its high sensibility.
Colorectal cancer (CRC) is one of the most common malignant tumors in humans beings. In the last decades
many experimental models have been developed in animals based in the use of chemical composites to
induce the formation and development of these tumors, many of them present similar characteristics to those
of natural occurrence aiming to the attainment of information on genesis, evolution, as well as diagnosis and
more efficient therapies for treating these neoplasias. Amongst the most used chemical composites is the 1,2-
dimetilhydrazine (DMH) because its morphological and histological similarity to those tumors. This study aims
to compare in vivo normal colon tissue and tumoral colon tissue, induced by DMH, in rats by near-infrared
Raman spectroscopy to permit the use in the near future for an efficient diagnosis in real time besides being
useful as an auxiliary method for several therapies, including the photodynamic therapy.
Photodynamic therapy (PDT) has been widely studied in the last decades and it is becoming a promising tool in the treatment of tumors of many kinds. PDT is based on photoactivation of a sensitized drug that is restrained in the tumor cells, producing highly reactive species that can destroy tumoral cells with minimum collateral effect. This study aimed to evaluate the effect of the PDT in induced neoplasias of the colon by 1,2-dimetilhidrazine in rats, using as photosensitizing drug the chloroaluminum phthalocyanine incorporated to the liposomes and to compare the methods of irradiation using continuous or fractionated energy in PDT. Ten Wistar rats were distributed randomly in 3 groups (G1, G2 and C), anaesthetized and submitted to PDT with of fractionated (G1) or continuum (G2) irradiation energy using as a source of excitement an InGaAl laser. After 3 hours of the laser irradiation, 2 animals of the G1 group, 2 animals of the G2 group and 1 animal of C group were sacrificed and samples of tumoral tissue were collected for histological analysis; the same procedure was carried through 24 hours after irradiation. There were no significant differences between the extensions of the induced areas of necrosis for PDT in the groups under fractionated or continuous irradiation for the parameters used in this study. New studies must be carried through, using different parameters and intervals of laser irradiation, aiming to maximize the effect of the PDT for the treatment of colon tumors.
The Transmissible Venereal Tumor (TVT) is a very common neoplasic disease in a free-roaming dogs which affects the extern genital and presenting resistance to conventional drugs that promote high toxicity. Photodynamic Therapy (PDT) is based in tumor cells irradiation after absorption of photosensitizer substance. At present, the protoporphirin IX (PP IX) has been explored in PDT due to be endogen, then it does not present toxicity effect. This substance can be obtained by exogenous way through aminolevulinic acid (5-ALA) administration in patient. The aim of this work was establish the optimal conditions for PDD (Phodynamic Diagnosis) to irradiate the tumor after ALA administration through fluorescence spectroscopy to improve the results with PDT. In this research was studied the 5-ALA 20% absorption in TVT of vaginal and penial mucous of a female and a male dog, respectivaly. This drug was administrated topically and after 30 minutes the fluorescence spectra were collected in intervals of 15 minutes during 120 minutes. The results showed that the maximum peak of PP IX in the tumor was between 60 and 105 minutes after the ALA application. In conclusion, the optimum effect will be achieved irradiating the tumor tissue into this period.
Healing is important for the success of the insertion of implants and for treating traumatic or pathologic injuries of the bone. Lasertherapy has been suggested as a mean of improving bone healing. Near infrared Raman spectroscopy was used to assess the amount of both inorganic and organic components of irradiated and control bone around dental implants inserted in to the tibia. Fourteen rabbits received a titanium implant on the tibia; eight of them were irradiated with λ830nm laser (Thera Lase, DMC, Sao Carlos, SP, Brazil, 21.5 J/cm2, 10mW, spot size 0.60mm) and six acted as controls. The animals were sacrificed 15, 30 and 45 days after the surgery, and specimens were prepared for Raman spectroscopy, which was collected at every four points from each three thirds of the bone around the implants. The results showed significant differences in the concentration of inorganic components in irradiated specimens between 15 and 30days (p < 0.05), 15 and 45 (p < 0.01); between irradiated and controls 30 and 45 days after surgery (p <0.01).
Concentration of organic components was also significantly different between irradiated and controls in periods of 30 to 45 days after surgery. It is concluded that LLLT does improve bone healing and Raman Spectroscopy can safely assess this.
Healing is important for the success of the insertion of implants and for treating traumatic or pathologic injuries of the bone. Lasertherapy has been suggested as a mean of improving bone healing. Raman spectroscopy was used to assess the amount of both inorganic and organic components of irradiated and non-irradiated bone around dental implants inserted in to the tibia. Fourteen rabbits received a titanium implant on the tibia, eight of them were irradiated with 830nm laser and six acted as controls. The animals were sacrificed 15, 30 and 45 days after the surgery, and specimens were prepared for Raman spectroscopy, which was collected at every four points from each three thirds of the bone around the implants. The results showed significant differences in the concentration of inorganic components in irradiated specimens between 15 and 30days, 15 and 45; between irradiated and controls 30 and 45 days after surgery. Concentration of organic components was also significantly different between irradiated and controls in periods of 30 to 45 days after surgery. It is concluded that LLLT does improve bone healing and this can be safely assessed by Raman Spectroscopy.
Comparative studies were carried out on the Al- phthalocyanine and Zn-phthalocyanine uptake in iliac artery wall of rabbit, through transadventitial fluorescence measurements. Two 3.5 Kg Norfolk make rabbit were used. The iliac artery was dissected, isolated and clamped at two points, 1.5 cm apart. The experiment comprised three steps, collecting fluorescence spectra at several point inside and outside of the clamped region of the artery. The excitation source was a 488 nm Argon Laser. Firstly, it was measured the pre and post clamping transadventitial fluorescence without the photosensitizer. After that, 50 (mu) L of AlPc or ZnPc solutions in Cremiphor EL, 2.5 (mu) M, were injected locally into the artery clamped region of each rabbit and fluorescence spectra were collected at t equals 0 and 15 min. Finally, elapsed 30 min. Temporal evolution of the drug uptake by the artery wall was studied analyzing the intensities of the fluorescence peaks at 682 and 678 nm for AlPc and ZnPc, respectively. It was found that AlPc is more absorbed and remains for a longer time in the iliac artery wall than the ZnPc.
Near-infrared (NIR) Raman spectroscopy has been studied for the last years for many biomedical applications. It is a powerful tool for biological materials analysis. Toxoplasmosis is an important zoonosis in public health, cats being the principal responsible for the transmission of the disease in Brazil. The objective of this work is to investigate a new method of diagnosis of this disease. NIR Raman spectroscopy was used to detect anti Toxoplasma gondii antibodies in blood sera from domestic cats, without sample preparation. In all, six blood serum samples were used for this study. A previous serological test was done by the Indirect Immunoenzymatic Assay (ELISA) to permit a comparative study between both techniques and it showed that three serum samples were positive and the other three were negative to toxoplasmosis. Raman spectra were taken for all the samples and analyzed by using the principal components analysis (PCA). A diagnosis parameter was defined from the analysis of the second and third principal components of the Raman spectra. It was found that this parameter can detect the infection level of the animal. The results have indicated that NIR Raman spectroscopy, associated to the PCA can be a promising technique for serological analysis, such as toxoplasmosis, allowing a fast and sensitive method of diagnosis.
A growing number of studies have demonstrated that autofluorescence and induced fluorescence spectroscopy can be used to distinguish normal and abnormal tissues in vivo. Through photochemical accumulation in atherosclerotic lesions it is possible to obtain induced fluorescence, this accumulation has been demonstrated in many researches. The aim of this study was to diagnose, by aluminum phthalocyanine transadventitial fluorescence, atherosclerotic plaques in artery wall of rabbits. Five male Norfolk rabbits were submitted to a high level cholesterol diet for 9 weeks. When the blood cholesterol level reached around 1000 mg/dl three of these animals were injected intravenously 50(M/50(l of aluminum phthalocyanine (AlPc). After 24 hours these animals were submitted to a surgery to expose the iliac artery. The transadventitial fluorescence spectra were acquired in several points over the artery. Characteristic peaks of the collagen, the porphyrin and the induced by AlPc were observed.
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