L. G. Marcassa

Detection of mechanical and disease stresses in citrus plants by fluorescence spectroscopy

We have investigated the detection of mechanical and disease stresses in citrus plants (Citrus limonia [L.] Osbeck) using laser-induced fluorescence spectroscopy. Due to its economic importance we have chosen to investigate the citrus canker disease, which is caused by the Xanthomonas axonopodis pv. citri bacteria. Mechanical stress was also studied because it plays an important role in the plants infection by such bacteria. A laser-induced fluorescence spectroscopy system, composed of a spectrometer and a 532 nm 10 mW excitation laser was used to perform fluorescence spectroscopy. The ratio of two chlorophyll fluorescence bands allows us to detect and discriminate between mechanical and disease stresses. This ability to discriminate may have an important application in the field to detect citrus canker infected trees.

Fluorescence spectroscopy applied to orange trees

In this work, we have applied laser-induced fluorescence spectroscopy to investigate biological processes in orange trees (Citrus aurantium L.). We have chosen to investigate water stress and Citrus Canker, which is a disease caused by the Xanthomonas axonopodis pv. citri bacteria. The fluorescence spectroscopy was investigated by using as an excitation source a 442-nm 15-mW HeCd gas multimode discharge laser and a 532-nm 10-mW Nd3+:YAG laser. The stress manifestation was detected by the variation of fluorescence ratios of the leaves at different wavelengths. The fluorescence ratios present a significant variation, showing the possibility to observe water stress by fluorescence spectrum. The Citrus Canker’s contaminated leaves were discriminated from the healthy leaves using a more complex analysis of the fluorescence spectra. However, we were unable to discriminate it from another disease, and new fluorescence experiments are planned for the future.

Detection of citrus canker in citrus plants using laser induced fluorescence spectroscopy.

Citrus canker is a serious disease caused by Xanthomonas citri subsp. citri bacteria, which infects citrus plants (Citrus spp.) leading to a large economic loss in citrus production worldwide. In Brazil citrus canker control is done by an official eradication campaign, therefore early detection of such disease is important to prevent greater economic losses. However, detection is difficult and so far it has been done by visual inspection of each tree. Suspicious leaves from citrus plants in the field are sent to the laboratory to confirm the infection by laboratory analysis, which is a time consuming. Our goal was to develop a new optical technique to detect and diagnose citrus canker in citrus plants with a portable field spectrometer unit. In this paper, we review two experiments on laser induced fluorescence spectroscopy (LIF) applied to detect citrus canker. We also present new data to show that the length of time a leaf has been detached is an important variable in our studies. Our results show that LIF has the potential to be applied to citrus plants.

Characterization of Light Penetration in Rat Tissues

OBJECTIVE The goal of this study is to determine the optical properties of different rat tissues with respect to spatial intensity variation and light distribution. We are interested mainly in the wavelength of 630 nm. Nevertheless, for liver tissue we have used 514 nm and 670 nm as well. BACKGROUND DATA In the past, many articles have been written about the interaction of lasers with rat tissues. However, the technique of imaging the light distribution allows us to obtain the spatial scattering as well as an effective attenuation coefficient for the light intensity. METHODS Slices of different rat tissues were placed between two microscope slide mounts (spaced by 3 mm). A laser beam was irradiated on the sandwiched tissue. A CCD camera placed on the side, orthogonal to the beam path, recorded the intensity distribution of the scattered light. Analysis of this spatial intensity profile allowed determining the variation of the intensity as the light penetrates the tissue. RESULTS We have found that abdominal wall fat presents the lowest exponential decay when compared with liver, muscle, and kidney. The obtained values provided good data about the light distribution in those tissues when irradiated with a nondiffuse laser beam. For all tissues, we observed a spherical light distribution and exponential decay. Cirrhotic liver shows much stronger decay than healthy liver. These results are useful for several applications of laser for biostimulation a phototherapy.

PDT experience in Brazil: A regional profile.

The success of PDT and its establishment into the existent hall of therapeutic modalities depends on the collection of reported experiences from around the world. In that sense, it is important to report approaches taken by different countries and what their views are on the future of PDT. Following this idea, we present our clinical experience in photodynamic therapy (PDT) in Brazil, as well as the experimental advances coming up in parallel with clinical implementation. This report is a consequence of pioneering work in a collaborative program involving the Physics Institute in São Carlos, São Paulo State (SP), Brazil, the Medical School of the University of São Paulo, Ribeirão Preto, SP, Brazil and the Cancer Hospital Amaral Carvalho, Jaú, SP, Brazil. This collaborative program, begun in 1997, with the first patient treated in 1999, has treated over 400 patients by late 2004. About 80% of lesions were located in the head and neck or skin, but experience is being built in esophagus, bladder, gynecology, and cutaneous recurrence of breast cancer, among others. The overall results have shown to be compatible with previously reported data. Modifications, whose goal is to improve patient benefit and optimize results, are being implemented as we gain experience. In parallel with the clinical development, several laboratories have started studying experimental whose purpose is to analyze the clinical results and to contribute to the worldwide effort to bring PDT to the forefront of therapies offered to patients. We present the overall results of our 5 years experience as well as the whole implementation process.

A Comparative Study on Application of Computer Vision and Fluorescence Imaging Spectroscopy for Detection of Huanglongbing Citrus Disease in the USA and Brazil

The overall objective of this work was to develop and evaluate computer vision and machine learning technique for classification of Huanglongbing-(HLB)-infected and healthy leaves using fluorescence imaging spectroscopy. The fluorescence images were segmented using normalized graph cut, and texture features were extracted from the segmented images using cooccurrence matrix. The extracted features were used as an input into the classifier, support vector machine (SVM). The classification results were evaluated based on classification accuracies and number of false positives and false negatives. The results indicated that the SVM could classify HLB-infected leaf fluorescence intensities with up to 90% classification accuracy. Though the fluorescence intensities from leaves collected in Brazil and the USA were different, the method shows potential for detecting HLB.

Enhancement of liver regeneration by the association of Hyptis pectinata with laser therapy.

Since new molecules that normally would accelerate regeneration can also be potentialized by light, the use of new substances combined with laser therapy seems to be a natural type of experiment. Therefore, the purpose of this study was to assess the effects of Hyptis pectinata leaves on liver regeneration after partial hepatectomy (PH) associated with laser therapy. Twenty-four rats were divided into four groups—PH(control), PHL (laser therapy), PH200 (200 mg/kg of Hyptis pectinata), and PHL200 (200 mg/kg of the plant and laser)—which were submitted to 67% hepatectomy. Laser treatment consisted of focusing the light on the remaining liver after hepatectomy. The data analyzed were serum levels of aminotransferases, liver regeneration, and mitochondrial function. Group PH200 showed a statistically significant decrease in AST levels, and PHL200 disclosed an augmentation in ALT levels. The liver regeneration index was significantly increased in group PHL200. Concerning liver mitochondrial respiratory assay, groups PH200 and PHL200 showed lower state 3 levels than groups PH and PHL. Group PHL showed an increase in state 4 levels and a reduction in membrane potential and RCR. The present study shows that the association of the aqueous extract of Hyptis pectinata leaves at 200 mg/kg with intraoperative laser therapy can stimulate liver regeneration and cause a reduction in liver mitochondrial respiratory function without altering its phosphorylative activity.

Detection of citrus canker and Huanglongbing using fluorescence imaging spectroscopy and support vector machine technique

Citrus canker and Huanglongbing (HLB) are citrus diseases that represent a serious threat to the citrus production worldwide and may cause large economic losses. In this work, we combined fluorescence imaging spectroscopy (FIS) and a machine learning technique to discriminate between these diseases and other ordinary citrus conditions that may be present at citrus orchards, such as citrus scab and zinc deficiency. Our classification results are highly accurate when discriminating citrus canker from citrus scab (97.8%), and HLB from zinc deficiency (95%). These results show that it is possible to accurately identify citrus diseases that present similar symptoms.

Effects of electric fields on ultracold Rydberg atom interactions

The behaviour of interacting ultracold Rydberg atoms in both constant electric fields and laser fields is important for designing experiments and constructing realistic models of them. In this paper, we briefly review our prior work and present new results on how electric fields affect interacting ultracold Rydberg atoms. Specifically, we address the topics of constant background electric fields on Rydberg atom pair excitation and laser-induced Stark shifts on pair excitation.

Magneto-optical trap for sodium atoms operating on the D1 line

Abstract We report on the observation of magneto-optical trapping of sodium atoms in the 3 S 1 2 → 3 P 1 2 transition (D 1 line). There are four combinations of atomic transitions within the D 1 line which produce trapping: two of them are achieved with the usual orthogonal circular polarizations used in the trap operating on the D 2 line, while the other two are just observed with reversed polarizations. The need of reversing the polarizations is attributed to the negative Zeeman shift of the lower hyperfine levels of the D 1 transition in alkalis. We have confined 10 5 to 10 7 atoms, depending on the atomic transitions involved.