Distinction of malignant melanoma and epidermis using IR micro-spectroscopy and statistical methods.

Infrared spectroscopy is widely perceived as a future technology for cancer detection and grading. Malignant melanoma, an aggressive skin cancer, is accessible to non-invasive IR radiation based surface probes for its identification and grading. The present work examines the differences in the IR spectra of melanoma tissues and the surrounding epidermis in skin biopsies with the objective of identifying diagnostic parameters and suitable computational/statistical methods of analysis. Melanoma could be differentiated from the epidermis in biopsies of 55 patients, using parameters derived from absorbance bands originating from molecular vibrations of nucleic acids and/or their bases. Additionally, absorbances from tyrosine and phosphate that are abnormally elevated in malignant melanoma could be used as markers. Two-dimensional plots of these parameters in tandem with advanced statistical methods successfully demonstrate the potential of IR spectroscopy to distinguish between epidermal and melanoma regions with a high classification success. The work underlines the importance of developing data analysis methods in FTIR based diagnosis using melanoma as a model system.

Assessment of drain water receiving effluent from tanneries and its impact on soil and plants with particular emphasis on bioaccumulation of heavy metals.

In the present study, impact of tannery and other industrial effluents on the physico-chemical characteristics of loamy drain water and their consequent impact on soil and plants irrigated with effluent have been studied. The study reveals most of the parameter pH, BOD5 and COD at sampling station I was higher than station II. Waste water quality at both Stations I and II exceeded prescribed limits (BIS) for safe disposal of effluents into the surface water Samples of soil and vegetables from the land irrigated with loamy drain water has been collected and analyzed for Cu, Zn, Ni, Cr Pb and Cd. The different metals showed different enrichment factor for loamy drain water irrigated soil and are as follows: Cd 30% (max), Pb 26%, Zn 18%, Cr 5%, Cu 5%, Ni 2% (min). For plant samples collected at polluted sites are Ni 46% spinach (whole plant) (max), Zn 42% spinach (whole plant), Cr 39% spinach (whole plant), Cu 33% spinach (whole plant), Pb 20% potato tuber, Cd 20% potato tuber (min). The levels of Zn 145, Cu 5.25, and Ni 39.25 microg/ g in spinach, Pb 29.25, Cr 38. 25 and Cd 3.2 microg/g in potato tuber grown on polluted soil irrigated with contaminated drain water were found more than the reference value, which may create chronic health hazard problem to human and cattle through food chain in long run. Accumulation of toxic heavy metals may be build up in the agriculturally productive land where it is treated with contaminated effluent enrich with metals in turn bio-concentrated in the edible fodder/plants.

Use of FTIR spectroscopy to distinguish between capsular types and capsular quantities in Streptococcus pneumoniae.

Fourier transform infrared (FTIR) spectroscopy has shown remarkable ability in distinguishing between bacterial species and identifying bacterial colony structures, when used in tandem with methods such as cluster analysis, principal component analysis, or linear discriminant analysis. The present work was aimed to evaluate the potential of FTIR-microscopy (FTIR-MSP) to distinguish between different serotypes and capsular quantities of Streptococcus pneumoniae. In general, the results obtained have consistently proven that the spectral information at the region 900-1,185 cm(-1) was sufficient to distinguish between various pneumococcal serotypes. Moreover, the method was able to differentiate between S. pneumoniae phase variants on the basis of their relative carbohydrate content. The unsupervised cluster analysis of the samples showed differences, not only in the carbohydrate content, but also in the region 1,350-1,480 cm(-1), which is dominated by absorptions due to lipids and phospholipids. This approach proved to be useful for the distinction between S. pneumoniae serotypes and between phase variants, which were shown to acquire different pathogenic capacity.

Can Fourier transform infrared spectroscopy at higher wavenumbers (mid IR) shed light on biomarkers for carcinogenesis in tissues?

Fourier transform infrared microspectroscopy (FTIR-MSP) has shown promise as a technique for detection of abnormal cell proliferation and premalignant conditions. In the present study, we investigate the absorbance in the sensitive wavenumber region between 2800 and 3000 cm(-1), which has been known to be due to the antisymmetric and symmetric stretching vibrations of CH2 and CH3 groups of proteins and lipids. We report common biomarkers from this region that distinguish between normal and malignant tissues and cell lines. Based on our findings, we propose that the wavenumber region around 2800 to 3000 cm(-1) in the FTIR spectra of cells and tissues could provide valuable scientific evidence at the onset of premalignancy and may be used for ex vivo and in vitro detection of carcinogenesis. To further examine the utility of these markers in cancer diagnosis and management, they are tested successfully in monitoring the changes occurring in leukemia patients during chemotherapy.

Distinction of cervical cancer biopsies by use of infrared microspectroscopy and probabilistic neural networks.

Fourier-transform infrared spectroscopy has shown alterations of spectral characteristics of cells and tissues as a result of carcinogenesis. The research reported here focuses on the diagnosis of cancer in formalin-fixed biopsied tissue for which immunochemistry is not possible and when PAP-smear results are to be confirmed. The data from two groups of patients (a control group and a group of patients diagnosed with cervical cancer) were analyzed. It was found that the glucose/phosphate ratio decreases (by 23-49%) and the RNA/DNA ratio increases (by 38-150%) in carcinogenic compared with normal tissue. Fourier-transform microspectroscopy was used to examine these tissues. This type of study in larger populations may help to set standards or classes with which to use treated biopsied tissue to predict the possibility of cancer. Probabilistic neural networks and statistical tests as parts of these biopsies predict the possibility of cancer with a high degree of accuracy (> 95%).

Fourier transform infrared spectroscopy in cancer detection.

The rapid developments in the field of infrared spectroscopy in the past decade have demonstrated a potential for disease diagnosis using noninvasive technologies. Several earlier studies have highlighted the advantage of using infrared spectroscopy both in the near- and mid-infrared regions for diagnostic purposes at clinical levels. The areas of focus have been the distinction of premalignant and malignant cells and tissues from their normal state using specific parameters obtained from Fourier transform infrared spectra, making it a rapid and reagent-free method. While it still requires pilot studies and designed clinical trials to ensure the applicability of such systems for cancer diagnosis, substantial progress has been made in incorporating advances in computational methods into the system to increase the sensitivity of the entire setup, making it an objective and sensitive technique suitable for automation to suit the demands of the medical community. The development of fiber-optics systems for infrared spectroscopy have further opened up new and modern avenues in medical diagnosis at various levels of cells, tissues and organs under laboratory and clinical conditions.

Characteristic absorbance of nucleic acids in the Mid-IR region as possible common biomarkers for diagnosis of malignancy.

FTIR spectroscopy has been extensively used to understand the differences between normal and malignant cells and tissues. In the present study, FTIR microspectroscopy was performed on biopsies to evaluate parameters deduced from changes in nucleic acid absorbance monitored at various characteristic wavenumbers in the Mid-IR region. The data showed that there were differences in the spectra of normal and malignant tissues from several organs such as colon, cervix, skin and blood with respect to absorbance due to nucleic acids. Similar results were observed in the case of cell lines that were transformed to induce carcinogenesis. Of the several ratios examined for consistency in differentiating cancer and normal tissues, the I(996 cm(-1))/I(966 cm(-1)) showed promise as a distinguishing parameter and was comparable to the I(1121 cm(-1))/I(1020 cm(-1)) ratio reported in many earlier studies. The absorbance of nucleic acids is presented with an emphasis on the application of FTIR microspectroscopy for diagnosis of malignancy. Our results indicate that usage of nucleic acid absorbance yield statistically significant parameters, which could differentiate normal and cancerous tissues.

Possible common biomarkers from FTIR microspectroscopy of cervical cancer and melanoma.

Detection of malignancy at early stages is crucial in cancer prevention and management. Fourier transform infrared (FTIR) spectroscopy has shown promise as a non-invasive method with diagnostic potential in cancer detection. Studies were conducted with formalin-fixed biopsies of melanoma and cervical cancer by FTIR microspectroscopy (FTIR-MSP) to detect common biomarkers, which occurred in both types of cancer distinguishing them from the respective non-malignant tissues. Both types of cancer are diagnosed on skin surfaces. The spectra were analysed for changes in levels of biomolecules such as RNA, DNA, phosphates and carbohydrate (glycogen). Whereas carbohydrate levels showed a good diagnostic potential for detection of cervical cancer, this was not the case for melanoma. However, variation of the RNA/DNA ratio as measured at I(1121)/I(1020) showed similar trends between non-malignant and malignant tissues in both types of cancer. The ratio was higher for malignant tissues in both types of cancer.

Fourier transform infrared microspectroscopy as a quantitative diagnostic tool for assignment of premalignancy grading in cervical neoplasia.

The early diagnosis and proper identification of cervical squamous intraepithelial lesions plays an important role in a good prognosis for the patient. However, the present practice of screening based on PAP (Papanicolaou) smear and histopathology makes it tedious and prone to human errors. We assess the validity of FTIR microspectroscopy (FTIR-MSP) of biopsies as a method to properly assign the correct stage of premalignancy in patients with symptoms of cervical intraepithelial neoplasia. For the first time we evaluate the biopsies based on the FTIR spectra for different grades of neoplasia in tandem with probabilistic neural networks (PNNs) and histopathology. The results show that the grading of neoplasia based on FTIR-MSP and a PNN differentiates the normal from premalignant with a high level of accuracy. The false positive identification of the normal as cervical intraepithelial neoplasia 1 (CIN1), CIN2, and CIN3 patients is 9.04, 0.01, and 0.01%, respectively. The false negative identification of CIN2 patients as normal and CIN1 patients is 0.01 and 4.4%, respectively. Similarly, the false negative identification of CIN3 patients as normal, CIN1, and CIN2 is 0.14, 6.99, and 9.61%, respectively. The small errors encountered in the grading are comparable to current methods, encouraging advanced studies for the development of mechanized equipment for the diagnosis and grading of premalignant cervical neoplasia.

Detection of abnormal proliferation in histologically 'normal' colonic biopsies using FTIR-microspectroscopy.

BACKGROUND: Abnormal crypt proliferation and development in the colon has been associated with premalignant stages of colon cancer. Conventionally, molecular markers are used to detect abnormal crypt proliferation. METHODS: In the present work, feasibility studies of FTIR-MSP to distinguish between normal and abnormal crypts from colon biopsies that show normal histopathological features have been undertaken. RESULTS: The results indicate that abnormal crypts show deviations in the pattern of absorbance in the Mid IR region along the crypt height when compared with the normal crypts. The crypts could be empirically classified into three groups such as crypts having a normal absorbance pattern for all biochemical components, crypts with abnormal absorbance pattern for some biochemical components and crypts with completely abnormal absorbance pattern along the height for all or most biochemical components studied by FTIR. The utilization of FTIR-MSP is proposed for diagnosis of abnormal metabolism at the molecular level of histologically completely normal-looking crypts, especially from those biopsies that are taken from sites far away from cancer. CONCLUSIONS: This method could give rise to a reduction in false-negative results during examination of biopsies using the conventional histopathological methods. The present method may be complementary to existing methods for precise demarcation of the zone of colostomy prior to colon cancer surgery.

Virulence of Streptococcus pneumoniae may be determined independently of capsular polysaccharide.

Mice were inoculated intranasally with Streptococcus pneumoniae isolates of serotype 14 with different genetic backgrounds (14R, 14DW) and a capsular switch of 14R, strain 9VR (serotype 9V). Inoculation of the mice with 14R and 9VR resulted in 60% mortality. All the mice survived 14DW inoculation. No differences in lungs' bacterial loads were found 3 h following inoculation. Bacterial clearance of 5 logs was observed 48 h after inoculation with 14DW versus within 1 log 48 h after inoculation with 14R and 9VR. No significant differences in bacterial size or the capsular amount could be found between 14R and 14DW. We conclude that factor(s) in addition to the capsule, contribute to disease outcome.

Assessment of industrial effluent and its impact on soil and plants.

The present study deals with the assessment of industrial water of an electronic component manufacturing unit with electroplating and its subsequent effects on soil and plants receiving the effluent. The physico-chemical parameters of the effluent samples showed higher value than that of ground water. The treated effluent was within the permissible limit. Microtox test was conducted and determined the degree of toxicity of untreated, treated effluents as well as the water sample collected at effluent discharge point of receiving river (confluence point). The physico- mechanical parameters of the soil samples were not changed due to irrigation of the treated effluent, but the concentration of metals were comparatively higher than the control soil. Higher accumulation of metals was found in the plant parts in naturally growing weeds and cultivated crop plant irrigated with treated effluent. Elevated accumulation of metals in Eichhornia crassipes and Marsilea sp. growing along the effluent channel has been identified as a potential source of biomonitoring of metals particularly for Cu and Ca and can be utilised for the removal of heavy metal from wastewater.

Ni(II) induced changes in cell cycle duration and sister-chromatid exchanges in cultured human lymphocytes.

Investigations from our laboratory and others have shown that Ni(II) treatments of cultured human lymphocytes produced a relatively small but significant increase in SCE frequency. Based on the known effects of Ni(II) on DNA replication, we evaluated whether Ni(II) produced a cell cycle delay in lymphocytes. Human lymphocytes of three normal subjects were exposed to 5, 10, and 25 microM of NiSO4 in culture medium and scored for the percent of metaphases in the first (M1), second (M2), and third (M3) cell cycle for harvest times spaced every 4 h from 36 to 72 h after culture initiation. Cell cycle duration was studied using Tice's BISACK method with certain modifications. All three doses of NiSO4 caused a delay of nearly 1.5 h in the initiation of cell division, but only 25 microM NiSO4 caused a lengthening in the cell cycle time of nearly 4 h for completion of the first cycle. Only at the highest dose of Ni(II) was there a significant increase in the SCE frequency compared to the control. When the proliferation rate index (PRI) was examined, the effect of 5 or 10 microM Ni(II) was negligible while the 25 microM concentration caused a suppression in the proliferation rate. The effect of Ni(II) on the cell cycle was much more pronounced than on the PRI. A significant increase in SCE frequency was observed only for the concentration of Ni(II) that caused a pronounced cell cycle delay, a result that is consistent with prior studies showing higher SCE responses for chemical treatments that lengthen the cell cycle.

Effects of nickel sulfate, lead sulfate, and sodium arsenite alone and with UV light on sister chromatid exchanges in cultured human lymphocytes.

Sister chromatid exchanges (SCE) have been examined in human lymphocytes following in vitro treatments with metal salts, nickel sulfate, lead sulfate and sodium arsenite. All of the metal salts produced significant increases in the SCE frequencies over the levels for untreated lymphocytes. The SCE frequencies were also examined for metal treatments combined with ultraviolet light (200 ergs/mm2). For the lead treatments combined with the UV dose selected, an additive SCE response was observed compared to the SCE responses for UV or metal alone. The nickel and arsenite treatments combined with UV produced a less than additive SCE response for most concentrations tested. These results suggest that nickel or arsenite present in complex mixtures may reduce the SCE response to other compounds in the mixture normally capable of producing a much stronger SCE response and therefore lead to an underestimate of the effects of chemical exposure.

Cytogenetic hazards from agricultural chemicals. 4. Sequential screening in the barley progeny test for cytogenetic activity of some systemic fungicides and a metabolite.

Four systemic fungicides and a metabolite: Benomyl, Carbendazim, Thiophanate-methyl, Dexon and dimethyl phenylenediamine were screened in the barley progeny test for cytogenetic activity. All affected germination, seedling growth, mitotic and meiotic activity, pollen fertility and seedset in the M1 generation to different degrees; however, no chlorophyll mutations were induced and the M1 effect were much reduced in the M2 progeny. Though the results compare well with those from studies on mammalian cells, the use of these fungicides may not affect the plant breeder and the user of grain.

Genetic toxicological of some plant flavonoids by the micronucleus test.

4 important flavonoid compounds, namely quercetin, kaempferol, neohesperidin dihydrochalcone and rutin, were administered to male mice for the detection of gross chromosomal anomalies by the micronucleus test. The first 3 compounds were positively clastogenic to different extents, whereas the 4th was negative.

Cytogenetic hazards from agricultural chemicals. I. A preliminary study on the responses of root meristems to exotoxin from Bacillus thuringiensis a constituent of a microbial insecticide, thuricide.

It reported for the first time that the exotoxin, thuringiensin A, from Bacillus thuringiensis, a component of the insecticide thuricide, inhibits spindle and cytokinesis and induces micronuclei, chromocentric nuclei and minor deviations in spindle activity. The binucleate cells also undergo mitosis yielding biprophases and bimetaphases. Spindle seems to have been inhibited even in bimetaphase. Microtubular systems and chromosomes are implicated as the primary targets. Most effects resemble those of caffeine, colchicine, aminopyrin, chloral hydrate and vinblastine to different extents, and are therefore suggestive of the anti-neoplastic and mutagenic potentialties of the exotoxin. The extensive use of thuricide on crop plants, in view of its mutagenic potential, may be hazardous. The results also suggest that the exotoxin may be used as a pre-treating agent in chromosome analysis and as a candidate-tagging tool for synchronization and cell cycle analysis, besides its probable utility in studies on cancer cells.