Lung cancer detection by native fluorescence spectra of body fluids--a preliminary study.

Lung cancer takes a heavy toll every year, since the survival rate is not more than 15%. In this paper, we present results of a novel technique based on the autofluorescence of body fluids like blood plasma, acetone extract of cellular components, sputa and urine of lung cancer patients (N=27). A set of ratio parameters based on the fluorescence peaks of tryptophan and elastin, in plasma and sputum; flavin, NADH (reduced nicotinamide adenine dinucleotide) and porphyrin in urine; porphyrin alone in acetone extract of formed elements, were all evaluated. Similar sets of ratios were obtained for age adjusted normal controls (N=27) and all these ratios were given as inputs to multivariate (principle component and discriminant) analyses, which showed that the two groups could be classified with an accuracy of about 90%. Since the instrumentation involved was an ordinary steady state Xe lamp based spectrofluorometer, the technique is of significant advantage in screening and early detection of lung cancer in high risk population such as heavy smokers.

Fluorescence spectroscopy as a novel technique for premarital screening of sickle cell disorders.

Spectral diagnostic screening for sickle cell disease was carried out on volunteer blood samples (N = 100). The samples were subjected to different diagnostic methods including conventional complete blood count (CBC), hemoglobin electrophoresis (HBE) and spectral diagnosis. For the spectral diagnostic method, we discriminated three different characteristic spectral features. In total, 15 samples were sickle cell trait (SCT), 34 samples were sickle cell disease (SCD), and the rest of the samples (N = 51) were normal controls. The spectral discrimination of the three different sets of samples was distinguished on the quantification of fluorescent biomolecules such as tyrosine, tryptophan, NADH, FAD, and porphyrins. The results were compared with the conventional standard CBC and capillary electrophoresis findings. The spectral diagnosis method exhibited a sensitivity and specificity greater than 90 % for the tested samples. This technique requires only 5 mL blood samples, has an analysis time of 20 min, exhibits high accuracy and may be used in small clinics in remote villages.

Fluorescence spectral detection of acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML): A novel photodiagnosis strategy.

Acute lymphoblastic leukemia (ALL) is a cancer of the lymphoid line of blood cell, showing a rapid growth of lymphoblastic immature cells. Acute myeloid leukemia (AML) is a cancer of the myeloid line of blood cells. Early diagnosis is crucial for the effective treatment of these patients. The current standard procedure of diagnosis is extensive blood count analysis, microscopic morphological investigations, bone marrow biopsy and flow-cytometer which are all time- consuming and expensive. Here we demonstrate the advantage a new technique for the diagnosis of ALL and AML based on the fluorescence spectra of blood plasma and RBCs samples from the above patients. Based on the 85 patients analyzed the results reveal that our approach could discriminate the two malignancies unambiguously from the normal with 88 % sensitivity and 80 % specificity. The abnormal decrease in the level of amino acid, tryptophan and coenzyme NADH and abnormal increase in the other amino acid, tyrosine and another coenzyme FAD, (both in comparison to the normal control) act as malignancy indicative biomarkers. Since the contrast parameter between the normal and malignant samples is four- fold, the potential for early detection of leukemia is high. The instrumentation and technique are new and simple; hence may have significant supplementary or complementary value with the existing diagnostic methods.

Detection of hemophilia by fluorescence spectroscopy: A photodiagnosis approach.

Hemophilia (HP), the deficiency of clotting factors such as VIII (FVIII) and IX, is an inherited blood disorder which is due to the lack of clotting protein. Conventional techniques for detecting hemophilia are based on clotting factor tests such as hemostasis assays to determine various types of hemophilia and its severity. In the current study, we propose a new approach involving a spectral technique to discriminate normal controls from hemophilia patients with an accuracy above 80 %. This accuracy is calculated on the normalized relative intensity based on the light measurement of blood components by analyzing a certain set of fluorescent bio molecules. Our results indicated that Red Blood Cells (RBCs) show lower porphyrin content and enzyme deficiencies in hemophilia patients than in normal controls. Therefore, the spectral features in hemophilia patients are different from those of normal controls. Thus, the proposed technique is a good alternative for the detection of hemophilia patients when compared with conventional detection techniques.

Diagnosis of thalassemia using fluorescence spectroscopy, auto-analyzer, and hemoglobin electrophoresis - A prospective study.

BACKGROUND: Hemoglobinopathies (HgP) are prevalent in certain regions of the world. The World Health Organization estimated that 5% of the world's population is a carrier of the potentially pathological hemoglobin (Hb) gene. METHODS: This study aimed to compare the performance of fluorescence spectroscopy, a simple and inexpensive method, with that of conventional techniques for diagnosing thalassemia. The red blood cell (RBC) counts and levels of Hb, HbA, HbA2, and HbS were estimated via conventional methods of complete blood count and Hb electrophoresis to diagnose thalassemia. RESULTS: The RBCs and Hb, particularly the average values of HbA and HbA2, were lower in patients with thalassemia than in the normal controls. These hematologic parameters were also analyzed via fluorescence spectroscopybased on fluorescent biomolecules including tyrosine (275 nm), tryptophan (290 nm), nicotinamide adenine dinucleotide (NADH) (370 nm), flavin adenine dinucleotide (FAD) (450 nm), and porphyrin (585-635 nm). In thalassemia patients, all these parameters were above the normal range, primarily due to abnormal depression of NADH and elevation of FAD. CONCLUSION: Thalassemia canbe diagnosed via a fluorescent spectral method with an accuracy of 85% for blinded groups. This method may be useful for screening patients and reducing the cost of diagnosis in many rural countries.

Facile spectroscopy and atomic force microscopy for the discrimination of α and ß thalassemia traits and diseases: A photodiagnosis approach.

Thalassemia (Thal) is an inherited blood disorder endemic to the Mediterranean and Middle East (e.g., KSA and UAE). This disease is caused by defects in the synthesis of one or more hemoglobin chains in red blood cells (RBCs). Alpha (α) Thal is caused by a reduced or absent alpha globin segment. Similarly, beta (ß) Thal is caused by a defect in the beta globin segment. We divided the diseases into four groups: α Thal trait, α Thal disease, ß Thal trait, and ß Thal disease. The α or ß Thal traits are milder variants of these diseases and do not require treatment; but ß Thal disease (and to a lesser extent, α Thal) causes hemolytic anemia, splenomegaly, and bone deformities and requires repeated lifelong blood transfusions. This paper presents results regarding the identification of Thal variants using fluorescence spectroscopy of blood biomolecules and atomic force microscopy analysis of the morphologic features of red blood cells. The combined results provide new insights into the characteristics of these diseases. Furthermore, this study shows why ß Thal disease subjects are often transfusion-dependent, and α Thal disease subjects are only occasionally transfusion dependent.

Impact of Diabetes Mellitus on Human Erythrocytes: Atomic Force Microscopy and Spectral Investigations.

Diabetes mellitus (DM) is a common metabolic disease indicated by high sugar levels in the blood over a prolonged period. When left untreated, it can lead to long-term complications, such as cardiovascular disease, stroke, and diabetic retinopathy or foot ulcers. Approximately 415 million people (about 8.3% of the world's population) had diabetes worldwide in 2015, with 90% of the cases classified as Type 2 DM, which is caused by insulin resistance that arises mostly from being overweight and from a lack of exercise. DM affects every part of the body, including the erythrocytes. The aim of the present report is to gain insight into the damage done to the erythrocytes of patients classified with pre-diabetes and diabetes (plenty are found in the Kingdom of Saudi Arabia, a country where young people encompass a large segment of the population). The study presents results on the morphological analysis of erythrocytes by atomic force microscopy (AFM) and molecular investigations by fluorescence spectroscopy (FS). Our results indicate significant differences (in the morphology, size, and hemolytic end products) between the erythrocytes of diabetic patients (HbA1C, glycated hemoglobin, levels of 8⁻10%) and normal controls. It is well-known that DM and smoking are two major contributory factors for cardiovascular diseases (CVDs), and our observations presented in this study suggest that diabetes plays a relatively less damaging role than smoking for CVD.

A Novel Technique of Spectral Discrimination of Variants of Sickle Cell Anemia.

Sickle cell anemia (SCA) is an inherited blood disorder with worldwide incidence of 15%; out of this, it is found in up to 20% in countries like Kingdom of Saudi Arabia and Bahrain. The standard conventional method of detection is complete blood count (CBC) followed by hemoglobin electrophoresis or high-performance liquid chromatography (HPLC) or both. In this context, spectral detection of variants of sickle cell anemia (SCA) is an innovative technique, which when made accurate and reliable could be an effective alternative, since the instrumentation is compact (5 kg) and hence portable. This makes mass screening even in remote villages possible. In this paper, we give the essential aspects of fluorescent spectral features of sickle cell trait (SCT), sickle cell disease (SCD), beta (ß) thalassemia trait (BTT) + SCD, and beta (ß) thalassemia disease (BTD) + SCD. All the above four major variants could be discriminated among themselves and also from the normal control blood sample. All these analyses could be carried out with 5 ml of blood, in a time period of 10 minutes. The results of this paper give strong support for an alternative method, a spectral technique, for molecular-level diagnosis of sickle cell anemia and other closely related blood disorders.

Time Evolution of the Excimer State of a Conjugated Polymer Laser.

An excited dimer is an important complex formed in nano- or pico-second time scales in many photophysics and photochemistry applications. The spectral and temporal profile of the excimer state of a laser from a new conjugated polymer, namely, poly (9,9-dioctylfluorenyl-2,7-diyl) (PFO), under several concentrations in benzene were investigated. These solutions were optically pumped by intense pulsed third-harmonic Nd:YAG laser (355-nm) to obtain the amplified spontaneous emission (ASE) spectra of a monomer and an excimer with bandwidths of 6 and 7 nm, respectively. The monomer and excimer ASEs were dependent on the PFO concentration, pump power, and temperature. Employing a sophisticated picosecond spectrometer, the time evolution of the excimer state of this polymer, which is over 400 ps, can be monitored.

An Efficient Violet Amplified Spontaneous Emission (ASE) from a Conjugated Polymer (PFO-co-pX) in Solution.

The optical of conjugated polymer poly[(9,9-dioctylfluorenyl-2,7-diyl)-co-(2,5-p-xylene)] also known as [(PFO-co-pX), ADS145UV], dissolved in a few solvents, has been measured. The absorption, emission spectra, and quantum yield have been investigated by using a spectrophotometer and spectrofluorometer, respectively. The properties of Amplified Spontaneous Emission (ASE) of conjugated PFO-co-pX polymer under different conditions such as solvent type, concentration, and pumping energy have been examined by using the tripled frequency of a Nd:YAG laser as a source of pumping. The relation between output energy and pumping energy for the samples with different concentrations in three solvents has been studied. In addition, efficiency and optical gain from the ASE were measured. Additionally, the stability of this polymer as a laser material was investigated. Among the host of conjugated polymer lasers obtained by optical pumping, this conjugated polymer has exhibited only one ASE band under a wide range of concentrations and pump power values. This is most likely due to the steric hindrance by the copolymer segment pX (2,5-p-xylene). This investigation has provided new insights into the excited state dynamics of conjugated polymer materials and has shown that this new conjugated polymer is quite efficient in the violet region.

Smoking Induced Hemolysis: Spectral and microscopic investigations.

Smoking is one of the major causes of lifestyle associated mortality and morbidity such as cancer of the oral cavity and lungs, and also cardiovascular diseases. In this study, we have provided evidences for the smoking-induced hemolysis using two methods: spectra of blood components and atomic force microscopic analysis of surface morphology. A total of 62 subjects (control = 31; smoker = 31: 21 male; 10 female in each set) were considered for the study. The findings indicate that smoking leads to potholes on the surface, swelling of shape, rupturing of erythrocytes, removal of hematoporphyrin and flushing into the plasma as metabolites of the erythrocyte. The overall morphology of the erythrocytes of the smoker group appears more like a Mexican hat. The mean surface roughness was 5.5 ± 3 nm for the smoker group, but 1.2 ± 0.2 nm for the control group. Such damages might help the toxins, (CO, peroxidants, aldehydes etc.,) to gain easy access and get strongly absorbed by the hemoglobin, leading to enhanced rates of hemolysis as shown by the spectral features of metabolites. This indicates that the average life span of the smoker's erythrocytes is significantly less than that of the control group.

Relaxation Oscillation with Picosecond Spikes in a Conjugated Polymer Laser.

Optically pumped conjugated polymer lasers are good competitors for dye lasers, often complementing and occasionally replacing them. This new type of laser material has broad bandwidths and high optical gains comparable to conventional laser dyes. Since the Stokes' shift is unusually large, the conjugated polymer has a potential for high power laser action, facilitated by high concentration. This paper reports the results of a new conjugated polymer, the poly[(9,9-dioctyl-2,7-divinylenefluorenylene)-alt-co-{2-methoxy-5-(2-ethylhexyloxy)-1,4-phenylene}](PFO-co-MEH-PPV) material, working in the green region. Also discussed are the spectral and temporal features of the amplified spontaneous emissions (ASE) from the conjugated polymer PFO-co-MEH-PPV in a few solvents. When pumped by the third harmonic of the Nd:YAG laser of 10 ns pulse width, the time-resolved spectra of the ASE show relaxation oscillations and spikes of 600 ps pulses. To the best of our knowledge, this is the first report on relaxation oscillations in conjugated-polymer lasers.

Fluorescence spectral diagnosis of malaria: a preliminary study.

BACKGROUND: Malaria is the most common disease transmitted by the bite by an infected female anopheles mosquito and caused by the plasmodium parasite. It is mostly prevalent in subtropical regions receiving abundant rain and supporting copious mosquito breeding. This disease is generally detected by the microscopic examination of blood films or antigen based rapid diagnostic test. Only occasionally the parasite DNA is detected using polymerase chain reaction in certain advanced, expensive laboratories. METHODS: An innovative spectral detection method based on the fluorescence spectra of a set of blood plasma biomolecules [tyrosine, tryptophan, nicotinamide adenine dinucleotide (NAD), and flavin adenine dinucleotide (FAD)] and red blood cell (RBC)-associated porphyrin is being evolved by our group. RESULTS: The research so far has exhibited sensitivity and specificity values exceeding 90% based on the spectral features of blood components of 14 malaria patients and 20 numbers of age adjusted normal controls. The fluorescent biomolecules go out of proportion when the malarial parasite breaks down the hemoglobin of blood. CONCLUSION: This technique has the potential to be used as an alternative diagnostic procedure for malaria since the instrumentation involved is portable and inexpensive. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_182.

Fluorescence spectral classification of iron deficiency anemia and thalassemia.

Thalassemia (Thal), sickle cell anemia, and iron deficiency anemia (IDA) are the most common blood disorders in many parts of the world, particularly in developing countries like India and Bangladesh. The well-established diagnostic procedure for them is the complete blood count (CBC); however, there is substantial confusion in discrimination between Thal and IDA blood samples based on such CBC. We propose a new spectral technique for reliable classification between the above two anemias. This is based on the identification and quantification of a certain set of fluorescent metabolites found in the blood samples of patients of Thal and IDA.

Cervical cancer detection by time-resolved spectra of blood components.

Fluorescence spectral techniques are very sensitive, and hence they are gaining importance in cancer detection. The biomarkers indicative of cancer could be identified and quantified by spectral or time domain fluorescence spectroscopy. The results of an investigation of time-resolved spectra of cellular components of blood obtained from cervical cancer patients and normal controls are given. The cancer indicative biomarker in this paper is porphyrin; it has a fluorescence decay time of 60% more in samples of cancer patients than those of normal controls. Based on such measurements, a randomized set comprising samples from cancer patients and controls (N=27 in total) could be classified with sensitivity (92%) and specificity (86%).

Fluorescence spectra of blood and urine for cervical cancer detection.

In the current study, the fluorescence emission spectra (FES) and Stokes shift spectra (SSS) of blood and urine samples of cervical cancer patients were obtained and compared to those of normal controls. Both spectra showed that the relative intensity of biomolecules such as porphyrin, collagen, Nicotinamide adenine dinucleotide, and flavin were quite out of proportion in cervical cancer patients. The biochemical mechanism for the elevation of these fluorophores is not yet definitive; nevertheless, these biomolecules could serve as tumor markers for diagnosis, screening, and follow-up of cervical cancers. To the best of our knowledge, this is the first report on FES and SSS of blood and urine of cervical cancer patients to give a sensitivity of 80% and specificity of 78%.

Spectral discrimination of benign and malignant prostate tissues--a preliminary report.

In this preliminary report, benign (n=8) and malignant (n=5) prostate tissues, in vitro, have been taken through autofluorescence spectroscopy. Employing Stokes' shift spectra and fluorescence emission spectra as tools of analysis, we were able to discriminate the two sets of tissues with sensitivity and specificity in excess of 85%. When the excised prostate chips were scanned with a spatial resolution of 1 mm, the epicenter of malignancy also could be delineated.

A new lung cancer biomarker--a preliminary report.

OBJECTIVE: To investigate a unique biomarker from the blood plasma and sputum of lung cancer patients based on native fluorescence analysis of body fluids. BACKGROUND: Conventionally, biomarkers indicative of malignancy are identified by biochemical or biophysical processes. Most of the cancer biomarkers, often useful in monitoring disease progression, have sensitivity and specificity in the range of 60%. METHODS: We employed synchronous fluorescence excitation spectroscopy (SFXS) and fluorescence emission spectroscopy for the native fluorescence analysis of blood plasma of 32 normal controls, 32 patients with lung cancer, and 32 patients with other types of cancer. RESULTS: Based on the native fluorescence analysis of blood plasma and sputum, we found that the structural protein elastin, with an excitation peak at 327 nm and an emission peak at 405 nm, is an exclusive biomarker for lung carcinoma with 77% sensitivity and 83% specificity from plasma alone, 92.3% sensitivity and 100% specificity from plasma acetone extract alone, and 66% sensitivity and 100% specificity from sputum alone. CONCLUSION: In this preliminary report with a limited number of lung cancer patients, we have used SFXS of plasma and sputa as the basis for a new technique identifying elastin as an exclusive lung cancer biomarker. This technique has the potential to become a new protocol for rapid and cost-effective screening and diagnosis of lung cancer.

Cancer detection by native fluorescence of urine.

Because cancer is a dreaded disease, a number of techniques such as biomarker evaluation, mammograms, colposcopy, and computed tomography scan are currently employed for early diagnosis. Many of these are specific to a particular site, invasive, and often expensive. Hence, there is a definite need for a simple, generic, noninvasive protocol for cancer detection, comparable to blood and urine tests for diabetes. Our objective is to show the results of a novel study in the diagnosis of several cancer types from the native or intrinsic fluorescence of urine. We use fluorescence emission spectra (FES) and stokes shift spectra (SSS) to analyze the native fluorescence of the first voided urine samples of healthy controls (N=100) and those of cancer patients (N=50) of different etiology. We show that flavoproteins and porphyrins released into urine can act as generic biomarkers of cancer with a specificity of 92%, a sensitivity of 76%, and an overall accuracy of 86.7%. We employ FES and SSS for rapid and cost-effective quantification of certain intrinsic biomarkers in urine for screening and diagnosis of most common cancer types with an overall accuracy of 86.7%.