Exogenous ß-cyclocitral treatment primes tomato plants against drought by inducing tolerance traits, independent of abscisic acid.

Drought is the most devastating stress for crops. Intensity and duration of drought determine the magnitude of plant damage; similarly, plant ability to counteract drought determines its tolerance capacity. Recent studies revealed that exogenous apocarotenoid treatment confers abiotic stress tolerance to plants. However, much less is known about the role of ß-cyclocitral (ßCC), the major apocarotenoid, in drought tolerance. Here, we demonstrate ßCC's role in improving plants' tolerance against drought stress. Tomato (Solanum lycopersicum L.) plants were independently treated with water and ßCC and grown under either water-limited or irrigated conditions. The ßCC-treated drought-exposed (BD) and ßCC-treated irrigated (BH) plants were analysed for the major drought tolerance associated traits; water-treated drought-exposed (CD) and water-treated irrigated plants (CH) were used as controls. On exposure to drought, unlike controls, ßCC-treated plants showed no wilting, higher RWC and stomatal conductance, unchanged ABA levels and stomatal closure. The BD plants had increased photosynthesis, chlorophyll content and enhanced root, but not shoot, growth. In addition, ßCC treatment enhanced proline accumulation and activity of SOD in both drought-exposed and well irrigated plants. Taken together, ßCC was identified as a potential candidate that improves tomato osmolyte accumulation and superoxide elimination, independent of ABA, and prepares the plant for upcoming drought stress. Our results suggest that ßCC can be used to prime crops against drought stress.

Peripapillary schisis in open-angle glaucoma.

PurposeTo report clinical features, topographic findings and outcome of 10 eyes with peripapillary schisis in open-angle glaucoma.Patients and methodsA retrospective review of patients with open-angle glaucoma who were noted to have peripapillary schisis on optical coherence tomography (OCT) were included. Serial peripapillary and macula infrared and OCT images, visual acuity, visual fields, and schisis appearance were reviewed.ResultsTen eyes of nine patients with open-angle glaucoma were detected to have the presence of peripapillary schisis. Nerve fibre layer schisis was detected in all eyes and one eye had an associated macular schisis. None of the eyes had an acquired pit of the optic nerve or pathological myopia. The mean intraocular pressures at detection was 18.3±4.3 mm Hg and the schisis resolved in four eyes after a mean follow-up of 21.2±8.8 months. Visual field worsening was noted in 4 of the 10 eyes and the resolution of schisis resulted in significant reduction in the retinal nerve fibre layer (RNFL) thickness.ConclusionsPeripapillary schisis detected during the normal course of open-angle glaucoma can resolve spontaneously and rarely involves the macula. Its resolution leads to reduction in RNFL thickness; therefore, caution is advised while interpreting serial scans.

Hallermann-streiff syndrome.

Hallermann-Streiff syndrome (HSS) is a rare disorder characterized by dyscephalia, with facial and dental abnormalities. We report a 12-year-old female child who presented with abnormal facial features, dental abnormalities and sparse scalp hair.

Prevalence and etiology of subclinical mastitis among buffaloes (Bubalus bubalus) in Namakkal, India.

Milk samples from 206 apparently healthy buffaloes of marginal farmers maintained under the prevailing field conditions were screened for Subclinical Mastitis (SCM) to determine its prevalence and etiology by White Side Test (WST), California Mastitis Test (CMT), Somatic Cell Count (SCC) and Bacteriological examination. 26.20% of animals were positive for SCM in SCC and bacteriological examination. Prevalence in graded Murrah buffaloes were found to be higher compared to graded Surti and Non-descript breeds. The prevalence of SCM was highest in above 6th stage of lactation. Single quarter infection (51.85%) was more common compared to multiple quarter infection. Hind (83.34%) and left side (57.41%) quarters were more prone to SCM. The indirect tests such as WST and CMT were found to be closely agreement with SCC and bacteriological examination. Due to their efficacious, easy to perform and interpret, these tests can effectively be employed to detect SCM under field conditions. Somatic cell count of = 3,00,000/mL of milk was regarded as the upper limit for normal buffaloes. The SCC of = 3,00,000/mL of milk with positive bacterial growth were used to diagnose SCM in the present study. Staphylococcus sp. 25 (46.30%) was the most common pathogens isolated from SCM followed by Streptococcus sp. 11 (20.37%) and E. coli 06 (11.11%) of the 54 bacterial isolates. Monobacterial and mixed bacterial infections were observed in 47 (87.04%) and 7 (12.96%) cases respectively.

Side-viewing fiberoptic catheter for biospectroscopy applications.

Utilization of fiberoptic catheters can turn the Raman and fluorescence spectroscopy systems into powerful bio-medical diagnostic probes. An in vivo bio-chemical diagnosis of some important organs like the esophagus, intestine, lung branches, artery, etc., can be possible by developing fiber-probes with good signal collection capabilities, a good flexibility to scan different spatial regions of the sample and less background signals generated in the probes themselves. An in vivo diagnosis of endoluminal inner walls utilizing front-viewing catheters (FVC) is very difficult because the internal diameter of these organs do not allow (excitation and collection) flexibility to access the different spatial regions of the sample. In this work we have developed, different side-viewing catheter (SVC) probes with a very small distal tip (semi sphere, phi approximately 1.5 mm) and micro mirrors allow beam steering of the excitation and collected radiation at a 90 degree angle. Preliminary results of spectroscopic applications have been presented. Reflectance, fluorescence and Raman scattering measurements have been used to compare the efficiency of SVC with traditional FVC probes. The results demonstrate that the SVC probes not only exhibit more flexibility but also similar spectral characteristics and signal collection efficiencies in comparison with conventional FVC probes.

Intensities of E. coli nucleic acid Raman spectra excited selectively from whole cells with 251-nm light.

Escherichia coli bacteria in the logarithmic growth phase have been investigated by UV resonance Raman spectroscopy. Bacterial whole-cell Raman spectra excited at 251 nm reflect nearly exclusively the nucleic acid composition even though a very large fraction of the bacterial mass is composed of protein. It has been demonstrated that if bacteria are grown under controlled (logarithmic growth) conditions, which give rise to organisms of known average biochemical composition, the intensities of E. coli Raman spectra can be explained quantitatively from the knowledge of component nucleic acid base resonance Raman cross sections.

UV resonance Raman detection and quantitation of domoic acid in phytoplankton.

Cultures of the phytoplankton diatom, Pseudonitzschia multiseries, have been harvested under controlled growth conditions ranging from late logarithmic to late stationary phase (17-58 days). The amount of domoic acid (DA) present in the growth media and in the homogenized cells has been determined by HPLC. Defined samples of media, homogenized cells, whole cells, and whole cells in media have been laser excited at 251 nm for the purpose of selectively exciting intense UV resonance Raman spectra from DA in the samples. Neither media nor cell component spectra from algae seriously interfere with DA spectra. The spectral cross sections for the dominant 1652-cm-1 mode of DA have been determined for 242-, 251-, and 257-nm excitation. Maximum sensitivities are achieved with 251-nm excitation because cross sections for DA are a maximum, and interference from other algal components becomes very small. DA concentrations that have been determined with 251-nm excitation by resonance Raman methods correlate closely with values determined independently with HPLC, especially at higher DA concentrations. The UV resonance Raman analysis of DA in phytoplankton algae is shown to be very sensitive and quantitative as well as rapid and nonintrusive.

Prospects for in vivo Raman spectroscopy.

Raman spectroscopy is a potentially important clinical tool for real-time diagnosis of disease and in situ evaluation of living tissue. The purpose of this article is to review the biological and physical basis of Raman spectroscopy of tissue, to assess the current status of the field and to explore future directions. The principles of Raman spectroscopy and the molecular level information it provides are explained. An overview of the evolution of Raman spectroscopic techniques in biology and medicine, from early investigations using visible laser excitation to present-day technology based on near-infrared laser excitation and charge-coupled device array detection, is presented. State-of-the-art Raman spectrometer systems for research laboratory and clinical settings are described. Modern methods of multivariate spectral analysis for extracting diagnostic, chemical and morphological information are reviewed. Several in-depth applications are presented to illustrate the methods of collecting, processing and analysing data, as well as the range of medical applications under study. Finally, the issues to be addressed in implementing Raman spectroscopy in various clinical applications, as well as some long-term directions for future study, are discussed.

Analysis of nucleotides and aromatic amino acids in normal and neoplastic colon mucosa by ultraviolet resonance raman spectroscopy.

The objective of this study was to explore the potential of using ultraviolet resonance Raman (UVRR) spectroscopy to analyze normal and neoplastic colon tissue. Ultraviolet light at 251 nm, generated from the third harmonic of a Titanium:Sapphire laser, was used to irradiate the surfaces of surgically resected human colon specimens from six patients, five clinically diagnosed with adenocarcinoma, and one with familial adenomatous polyposis. All grossly neoplastic samples found to contain mucosal dysplasia or invasive adenocarcinoma upon histologic evaluation, were analyzed in parallel with normal tissue obtained from the same specimen and located at least 1 cm away from grossly neoplastic tissue. The colon spectra were modeled as a linear combination of nucleotide, aromatic amino acid, and lipid lineshapes, using chemical standards as a reference. Nucleotide and amino acid contributions to the UVRR spectra were quantified by a least squares minimization method. The least squares minimization spectral model was verified in aqueous solutions, where relative concentrations of free nucleotides and DNA were quantified with < 10% error. Of the 11 neoplastic samples studied from the 6 specimens, 10 showed either a lower amino acid/nucleotide ratio, a lower level of adenyl (A) signal, or both when compared with their normal counterpart. Lower amino acid/nucleotide ratio was present in five of six samples containing only dysplasia, and three of the five samples containing invasive adenocarcinoma. Lower A was present in all five samples containing invasive cancer, and in three of the six samples containing only dysplasia. This lower level of A corroborates previously published biochemistry work showing a lower level of total adenylates in tumor homogenates compared with normal tissue. Our data indicate that surface UVRR may provide unique information about site-to-site changes in cellular metabolites during colon carcinogenesis.

Diffuse reflectance spectroscopy of human adenomatous colon polyps in vivo.

Diffuse reflectance spectra were collected from adenomatous colon polyps (cancer precursors) and normal colonic mucosa of patients undergoing colonoscopy. We analyzed the data by using an analytical light diffusion model, which was tested and validated on a physical tissue model composed of polystyrene beads and hemoglobin. Four parameters were obtained: hemoglobin concentration, hemoglobin oxygen saturation, effective scatterer density, and effective scatterer size. Normal and adenomatous tissue sites exhibited differences in hemoglobin concentration and, on average, in effective scatterer size, which were in general agreement with other studies that employ standard methods. These results suggest that diffuse reflectance can be used to obtain tissue information about tissue structure and composition in vivo.

Spectroscopic characterization of a DNA-binding domain, Z alpha, from the editing enzyme, dsRNA adenosine deaminase: evidence for left-handed Z-DNA in the Z alpha-DNA complex.

Double-stranded RNA adenosine deaminase (ADAR1) is an ubiquitous enzyme in metazoa that edits pre-mRNA changing adenosine to inosine in regions of double-stranded RNA. Zalpha, an N-terminal domain of human ADAR1 encompassing 76 amino acid residues, shows apparent specificity for the left-handed Z-DNA conformation adopted by alternating (dGdC) polymers modified by bromination or methylation, as well as for (dGdC)13 inserts present in supercoiled plasmids. Here, a combination of circular dichroism, fluorescence, and gel-retardation studies is utilized to characterize recombinant Zalpha peptide and to examine its interaction with DNA. Results from laser-Raman spectroscopy experiments provide direct evidence for the existence of Z-DNA in peptide-DNA complexes.

Raman spectroscopy and fluorescence photon migration for breast cancer diagnosis and imaging.

We are developing optical methods based on near infrared Raman spectroscopy and fluorescence photon migration for diagnosis and localization of breast cancer. We demonstrate the ability of Raman spectroscopy to classify accurately normal, benign and malignant breast tissues, an important step in developing Raman spectroscopic needle probes as a tool for improving the accuracy of needle biopsy. We also show that photon migration imaging can be used to localize accurately small fluorescent objects imbedded in a thick turbid medium with realistic optical properties, thus demonstrating the potential of this technique for optical imaging.

Diagnosis of head and neck precancerous lesions in an animal model using fluorescence spectroscopy.

Laser-induced fluorescence (LIF) of tissues depends on their biochemical and histomorphologic characteristics. LIF spectroscopic properties of 9,10-dimethyl-1,2-benzanthracene (DMBA)-induced precancerous and early cancerous lesions in a hamster buccal pouch mucosa model were studied. Fluorescence spectra from neoplastic lesions showed a characteristic fluorescence peak in the red region of the visible spectrum centered between 630 and 640 nm when excited with 410-nm light. Using this as a diagnostic criterion, 45 of 49 lesions studied were correctly diagnosed, including early dysplastic lesions. Follow-up study of four dysplastic lesions over 2 weeks revealed an increase in red fluorescence intensity. The findings of these experiments suggest that LIF spectroscopy may be a valuable noninvasive technique not only for early diagnosis of head and neck cancer, but also to probe a possible biochemical surrogate biomarker in the follow-up of suspected lesions.

Spectral pathology.

We are investigating the use of optical spectroscopy (fluorescence, reflectance, Raman scattering) for detecting precancerous lesions in the mucosal linings of hollow organs. We present a morphological model for extracting quantitative pathological information from fluorescence spectra, using colonic dysplasia as an example. The potential of this technique in providing histological information in real time without the need for tissue removal is discussed.

Autofluorescence characteristics of oral mucosa.

BACKGROUND: The fluorescence characteristics of tissues depend upon their biochemical composition and histomorphological architecture, both of which undergo a change during malignant transformation. These changes are detectable as an alteration in the fluorescence spectral profile of the tissues. METHODS: Biopsy specimens from clinically suspicious lesions and normal-appearing oral mucosa were obtained from patients. Fluorescence spectroscopic measurements were obtained to study the differences between normal and dysplastic tissues and to determine the most appropriate excitation wavelength(s) for exploiting these differences. RESULTS: Fluorescence spectra from a total of 12 histologically normal (healthy mucosa or benign lesions) and ten abnormal (dysplastic or malignant) tissue samples were compared. Significant spectral differences were seen between the two groups. These differences were most marked at the excitation wavelength of 410 nm. Using this wavelength, fluorescence correctly diagnosed 20 of 22 samples studied. CONCLUSIONS: This technique accurately differentiates normal from abnormal tissues in vitro and has the potential applications for in vivo use as a noninvasive diagnostic tool.

Early diagnosis of upper aerodigestive tract cancer by autofluorescence.

OBJECTIVE: To explore the potential of autofluorescence spectroscopy as a tool for early detection of upper aerodigestive tract cancer. DESIGN: Autofluorescence spectral characteristics of 19 untreated oral and oropharyngeal lesions in 13 patients were studied with excitation wavelengths of 370 and 410 nm generated by a nitrogen pumped dye laser. Ten healthy volunteers were recruited to characterize the fluorescence spectra of normal mucosa at different oral sites and to study individual variations. Fluorescence intensity and line shape of the spectra from lesions were compared with the same parameters from the contralateral control site in the same individual. SETTING: Otolaryngology Research Center, Department of Otolaryngology-Head and Neck Surgery, New England Medical Center, Boston, Mass. RESULTS: The ratio of peak fluorescence intensities of the neoplastic lesions to contralateral normal control mucosa were consistently different compared with these ratios in benign lesions or normal mucosa. These differences were seen in 2 distinct regions of the fluorescence spectrum with both of the excitation wavelengths, but were more obvious with the excitation wavelength of 410 nm. Using these differences, we were able to correctly diagnose 17 of the 19 lesions studied, with 2 false-positive results. CONCLUSIONS: Neoplastic oral mucosa shows consistent differences in autofluorescence spectral intensity and line shape when compared with the normal mucosa in the same individual. These early results show that fluorescence spectroscopy may represent a useful technique for noninvasive early diagnosis of cancer of the upper aerodigestive tract.

Detection of dysplasia at colonoscopy using laser-induced fluorescence: a blinded study.

BACKGROUND: Laser-induced fluorescence spectroscopy has the potential to detect colonic dysplasia in vivo. However, previous studies have limited their analyses to multivariate regression techniques and unblinded retrospective evaluation. The purpose of this study was to develop a probability-based algorithm to detect colonic dysplasia using laser-induced fluorescence spectroscopy and to evaluate it in a blinded manner. METHODS: Fluorescence spectra were collected from normal mucosa and colonic polyps during colonoscopy using 370 nm excitation. Tissue was classified as normal, hyperplastic, or adenomatous by histologic examination. Preliminary data was used to devise an algorithm to differentiate tissue type based on probability distributions of the fluorescence intensity at 460 nm and the ratio of the intensity at 680 nm to that at 600 nm. The algorithm was then tested in a blinded fashion. RESULTS: The algorithm correctly determined the tissue type in 88% of cases, equal to the agreement of independent pathologists. Sensitivity, specificity, and positive predictive value for the detection of dysplasia was 90%, 95%, and 90%, respectively. CONCLUSIONS: Dysplasia was detected in vivo using fluorescence spectroscopy and a probability-based algorithm. This method may form the basis for a new surveillance technique for patients with increased risk for dysplastic transformation.

Morphological model of human colon tissue fluorescence.

Fluorescence spectroscopy of tissue is a promising technique for early detection of precancerous changes in the human body. Investigation of the microscopic origin of the clinically observed tissue fluorescence can provide valuable information about the tissue's histology. The objective of this study was the development of a morphological model of colon tissue fluorescence which connects the clinically observed spectra with their underlying microscopic origins. Clinical colon tissue fluorescence which connects the clinically observed spectra with their underlying microscopic origins. Clinical colon tissue fluorescence spectra were modeled by measuring the intrinsic fluorescence properties of colon tissue on a microscopic level and by simulating light propagation in tissue using the Monte-Carlo method. The computed spectra were in good agreement with the clinical spectra acquired during colonoscopy, and exhibited the characteristic spectral features of the in vivo collected spectra. Our analysis quantitated these spectral features in terms of the intrinsic fluorescence properties of tissue and its general histological characteristics. The fluorescence intensity difference between normal and adenoma observed in vivo was found to be due to the increased hemoglobin absorption, the reduced mucosal fluorescence intensity, and the absence of submucosal fluorescence in adenomatous polyps. The increased red fluorescence in adenoma was found to be associated with the dysplastic crypt cell fluorescence.

Rapid multiexcitation fluorescence spectroscopy system for in vivo tissue diagnosis.

We have designed, fabricated, and tested a compact, transportable, excitation-emission spectrofluorimeter with optical-fiber light delivery and collection for use in rapid analysis of tissues in a clinical setting. This system provides up to eleven different excitation wavelengths, permitting collection of all the corresponding emission spectra in approximately 600 ms. It uses a N(2) laser that pumps a sequence of dyes placed in cuvettes on a rotating wheel. A white-light excitation source permits acquisition of the tissue's diffuse reflectance spectrum on each cycle. Return fluorescence and reflected light are dispersed by a small spectrograph and detected by a photodiode-array detector. The system can collect a single-shot spectrum from biological tissue with a signal-to-noise ratio in excess of 50:1.

Espectrofluorímetro de multi-excitaçäo portátil para a rápida deteçäo de displasias / Spectrofluorimeter of portable multi-excitation for rapid detection of dysplasia

Neste trabalho é apresentado o projeto e desenvolvimento de um espectrofluorímetro para a obtenção de espectros de fluorescência e refletância difusa de tecidos biológicos em um tempo inferior a 1s. Para acessar a região da diagnose, o sistema utiliza um catéter a fibra óptica para excitação do tecido e captação da fluorescência emitida. O sistema é desenvolvido para aplicação em procedimentos clínicos in vivo, onde o tempo de realização do experimento é objetivo de extrema importância

Abstract- This work presents the project and development of a spectrofluorimeter in order to obtain the fluorescence spectra and diffuse reflectance from biological tissues with period of time below 1 s. The diagnose region is reached with an optical fiber catheter for tissue excitation and collection of the emitted fluorescence. This system is developed for in vivo clinicai applications, where duration of the experimental procedure is a very important parameter