Early and rapid detection of potato's fungal infection by Fourier transform infrared microscopy.

Fungi are considered serious pathogens to many plants and can cause severe economic damage. Early detection of these pathogens is very important and might be critical for their control. The available methods for detection of fungi are time consuming and not always very specific. Fourier transform infrared (FT-IR) microscopy has proved to be a reliable and sensitive method for detection of molecular changes in cells. Fungi pathogens display typical infrared spectra that differ from the spectra of substrate material such as potato. In the present study we used FT-IR microscopy for early and rapid detection of the potato fungal pathogen Colletotrichum coccodes on the surface of potato tubers. Infected potatoes with this fungal pathogen and uninfected potatoes were examined and correctly classified as infected or not infected by FT-IR microscopy at very early stages of infection when no morphological signs of infection could be seen. Unique spectral biomarkers were found in naturally infected potatoes compared to disease-free control potatoes.

Continuous monitoring of WBC (biochemistry) in an adult leukemia patient using advanced FTIR-spectroscopy.

Fourier transform infrared (FTIR)-spectroscopy has been found useful for monitoring the effectiveness of drugs during chemotherapy in leukemia patients. In the present work, spectral changes that occurred in the white blood cells (WBC) of an adult acute myeloid leukemia (AML) patient and their possible utilization for monitoring biochemistry of WBC were investigated. The phosphate absorbance from nucleic acids and the lipid-protein ratio in the WBC decreased immediately after treatment and then increased to levels of a control group. Similar observations were recorded in child patients with acute lymphoblastic leukemia (ALL) who were used as test cases. These parameters maybe used as possible markers to indicate successful remission and suggest that FTIR-spectroscopy may provide a rapid optical method for continuous monitoring or evaluation of a WBC population.

Use of Fourier transform infrared microscopy for the evaluation of drug efficiency.

Fourier transform infrared (FTIR) spectroscopy has been used by chemists as a powerful tool to characterize inorganic and organic compounds. In this study, we examine the potential of FTIR microspectroscopy for early evaluation of therapy efficiency. For this purpose, we examine the effect of acyclovir (a known antiherpetic drug) on the development of herpes simplex virus type 1 (HSV-1) infection in cell culture. Also, we examine spectral changes in lymphocytes obtained from leukemia patients after appropriate chemotherapy treatment. Our results show early and significant spectral indicators for successful infection of Vero cells with HSV-1. Treatment of these infected cells with increasing doses of acyclovir reduces clearly the spectral changes caused by the infection in a correlation with inhibiting the development of the cytopathic effect (CPE) induced by this virus. Also significant and consistent spectral differences between lymphocytes from human leukemia patients compared to that from healthy persons are obtained. Treatment of these leukemia patients with appropriate drugs reduces significantly these spectral differences in a correlation with the improvement of the patient's clinical situation. It seems that FTIR spectroscopy can be used as an effective tool for early evaluation of the efficiency of drugs.

FTIR microscopy as a method for identification of bacterial and fungal infections.

Fourier-transform infrared (FTIR) microscopy is considered to be a comprehensive and sensitive method for detection of molecular changes in cells. The advantage of FTIR microspectroscopy over conventional FTIR spectroscopy is that it facilitates inspection of restricted regions of the examined sample. In the present study, we examined the potential of FTIR microscopy as an easy, rapid and reliable technique for discrimination between bacteria and fungi both of which are involved in various human and other animal infections. In many cases, there is no easy and rapid technique for identifying the cause of such infections whether it is bacteria, fungi or both. Knowing such information in a rapid way could be highly important for effective therapy. Our results proved detectable and significant spectral differences between bacterial and fungal samples. Representative peaks of bacteria and fungi appeared clearly in the spectra of a mixture of bacteria and fungi. It seems that this technique could be used for rapid discrimination between bacterial and fungal infections and contaminations.

FTIR microscopy detection of cells infected with viruses.

Fourier-transform infrared (FTIR) microscopy is considered a comprehensive and sensitive method for detection of molecular changes in cells. The advantage of FTIR microspectroscopy over conventional FTIR spectroscopy is that it facilitates inspection of restricted regions of a cell culture or a tissue. We have shown that it is possible to apply FTIR microscopy as a sensitive and effective assay for the detection of cells infected with various members of the herpes family of viruses and retroviruses. Detectable and significant spectral differences between normal and infected cells were evident at early stages of the infection. Impressive changes in several spectroscopic parameters were seen in infected compared with uninfected cells. It seems that the change in spectral behavior is specific to the infecting virus, because cells infected with herpesviruses showed different spectral changes compared with cells infected with retroviruses.

The use of FTIR microscopy for evaluation of herpes viruses infection development kinetics.

The kinetics of Herpes simplex infection development was studied using an FTIR microscopy (FTIR-M) method. The family of herpes viruses includes several members like H. simplex types I and II (HSV I, II), Varicella zoster (VZV) viruses which are involved in various human and animal infections of different parts of the body. In our previous study, we found significant spectral differences between normal uninfected cells in cultures and cells infected with herpes viruses at early stages of the infection. In the present study, cells in cultures were infected with either HSV-I or VZV and at various times post-infection they were examined either by optical microscopy or by advanced FTIR-M. Spectroscopic measurements show a consistent decrease in the intensity of the carbohydrate peak in correlation with the viral infection development, observed by optical microscopy. This decrease in cellular carbohydrate level was used as indicator for herpes viruses infection kinetics. This parameter could be used as a basis for applying a spectroscopic method for the evaluation of herpes virus infection development. Our results show also that the development kinetics of viral infection has an exponential character for these viruses.

Studies on acute human infections using FTIR microspectroscopy and cluster analysis.

A novel methodology for the diagnosis of acute infections using FTIR microspectroscopy (FTIR-MSP) data on blood components and cluster analysis is presented. Blood samples were collected from 11 patients suffering from various infections and 16 age-matched healthy human controls. Blood components such as white blood cells, red blood cells, and plasma were isolated using standard procedures and FTIR-MSP of these components was utilized. A cluster analysis of the FTIR spectra was performed. The spectra obtained from the three blood components of patients were different from those of controls. The FTIR spectra of white blood cells from patients suffering infections were significantly different from the controls. Cluster analyses of averaged FTIR-MSP spectra of white blood cells provided 100% classification between patients and healthy controls.

Spectroscopic evaluation of the effect of a red microalgal polysaccharide on herpes-infected Vero cells.

The sulfated polysaccharide obtained from a species of red microalga has proved to be a potent antiviral agent against various members of the herpes family. In the present study, we used microscopic Fourier transform infrared spectroscopy (FT-IR) to investigate differences between normal cells, those infected with herpes viruses, and infected cells treated with red microalgal polysaccharide. FT-IR enables the characterization of cell or tissue pathology based on characteristic molecular vibrational spectra of the cells. The advantage of microscopic FT-IR spectroscopy over conventional FT-IR spectroscopy is that it facilitates inspection of restricted regions of cell cultures or tissue. Our results showed significant spectral differences at early stages of infection between infected and noninfected cells, and between infected cells treated with the polysaccharide and those not treated. In infected cells, there was an impressive decrease in sugar content and a considerable increase in phosphate levels in conjunction with the infection progress. Our results also proved that sugars penetrated and accumulated inside cells treated with the red microalgal polysaccharide. These could have been sugar fragments of low molecular weight present in the polysaccharide solution, despite purification by dialysis. Such sugar accumulation might be responsible for a breakdown in the internal steps of the viral replication cycle.

FTIR microspectroscopy of malignant fibroblasts transformed by mouse sarcoma virus.

Fourier transform infrared microspectroscopy (FTIR-MSP), which is based on the characteristic molecular vibrational spectra of cells, was used to investigate spectral differences between normal primary rabbit bone marrow (BM) cells and bone marrow cells transformed (BMT) by murine sarcoma virus (MuSV). Primary cells, rather than cell lines, were used for this research because primary cells are similar to normal tissue cells in most of their characteristics. Our results showed dramatic changes in absorbance between the control cells and MuSV124-transformed cells. Various biological markers, such as the phosphate level and the RNA/DNA obtained, based on the analysis of the FTIR-MSP spectra, also displayed significant differences between the control and transformed cells. Preliminary results suggested that the cluster analysis performed on the FTIR-MSP spectra yielded 100% accuracy in classifying both types of cells.

Spectroscopic characterization of human and mouse primary cells, cell lines and malignant cells.

Fourier transform infrared (FTIR) spectroscopy is currently being developed as a new optical approach to the diagnosis and characterization of cell or tissue pathology. The advantage of FTIR microspectroscopy over conventional FTIR spectroscopy in the diagnosis of malignancies is that it facilitates inspection of restricted regions of the cell culture or tissue. In this study, we set out to evaluate FTIR microspectroscopy as a diagnostic tool for identifying retrovirus-induced malignancies. Our study showed significant and consistent differences between cultures of different types of cells of both mouse and human origin, i.e. primary fibroblast cells (one to two passages in cell culture), fibroblast cell lines and malignant cells transformed by murine sarcoma virus. An impressive decrease in the levels of phosphate and other metabolites was seen in malignant cells compared with primary cells. The levels of these metabolites in the cell lines were significantly lower than in the primary cells but higher than in the malignant cells. In addition, the peak attributed to the PO2- symmetric stretching mode at 1082 cm(-1) in primary cells shifted significantly to 1085 cm(-1) for the cell line and to 1087 cm(-1) for the malignant cells. These differences taken together with differences in the shapes of various bands throughout the spectrum strongly support the possibility of developing FTIR microspectroscopy for the detection and study of malignant--and possibly premalignant--cells.

FTIR spectroscopic method for detection of cells infected with herpes viruses.

Microscopic FTIR spectroscopy was used to investigate the spectral differences between normal cells in culture and cells infected with various members of the herpes family of viruses [Herpes simplex (HSV) and Varicella zoster (VZV)]. The main objective of this study is to evaluate the possibility of developing microscopic FTIR spectroscopy as a sensitive assay for the detection of herpetic infections at their early stages. The advantage of this method over conventional FTIR spectroscopy is that it facilitates inspection of restricted regions of tissue. Our results showed significant and consistent differences between all normal and HSV or VZV infected cells that were tested. Detectable and significant spectral differences between normal and infected cells are seen as early as 24 h postinfection, but the damage of the cells (cytopathic effect), caused by the infecting virus, can be seen by optical microscope observations at only 3 days postinfection. An impressive increase in the levels of vital cellular metabolites was seen in the herpes virus infected cells compared to normal cells. It seems that this spectral behavior is unique for infection with herpes viruses, because when these cells were infected with other viruses from different families like retroviruses, a considerable decrease in the levels of vital cellular metabolites was seen in infected cells compared to normal cells. Cluster analysis performed on FTIR mass chromatography yielded 100% accuracy in classifying control uninfected and VZV or HSV infected cells. Our data strongly support the possibility of developing FTIR microscopy as a diagnostic method for early detection of herpetic infections.

Anti-viral activity of red microalgal polysaccharides against retroviruses.

Red microalgal polysaccharides significantly inhibited the production of retroviruses (murine leukemia virus- MuLV) and cell transformation by murine sarcoma virus(MuSV-124) in cell culture. The most effective inhibitory effect of these polysaccharides against both cell transformation and virus production was obtained when the polysaccharide was added 2 h before or at the time of infection. Although, addition of the polysaccharide post-infection significantly reduced the number of transformed cells, but its effect was less marked than that obtained when the polysaccharide was added before or at the time of infection.The finding that the inhibition of cell transformation by MuSV-124 was reversible after removal of the polysaccharide suggested that microalgal polysaccharides inhibited a late step after provirus integration into the host genome. In conclusion, our findings could support the possibility that the polysaccharide may affect early steps in the virus replication cycle, such as virus absorption into the host cells, in addition to its effect on a late step after provirus integration.