Early detection of colorectal cancer relapse by infrared spectroscopy in "normal" anastomosis tissue.

Colorectal cancer is one of the most aggressive cancers usually occurring in people above the age of 50 years. In the United States, colorectal cancer is the third most diagnosed cancer. The American Cancer Society has estimated 96,830 new cases of colon cancer and 40,000 new cases of rectal cancer in 2014 in the United States. According to the literature, up to 55% of colorectal cancer patients experience a recurrence within five years from the time of surgery. Relapse of colorectal cancer has a deep influence on the quality of patient life. Infrared (IR) spectroscopy has been widely used in medicine. It is a noninvasive, nondestructive technique that can detect changes in cells and tissues that are caused by different disorders, such as cancer. Abnormalities in the colonic crypts, which are not detectable using standard histopathological methods, could be determined using IR spectroscopic methods. The IR measurements were performed on formalin-fixed, paraffin-embedded colorectal tissues from eight patients (one control, four local recurrences, three distant recurrences). A total of 128 crypts were measured. Our results showed the possibility of differentiating among control, local, and distant recurrence crypts with more than a 92% success rate using spectra measured from the crypts' middle sites.

Contamination of contact lens storage cases of asymptomatic refractive surgery candidates.

PURPOSE: To examine the rates of contamination of contact lens storage cases of refractive surgery candidates and compare the growth yields of the traditional method of culturing and a broth-based method. METHODS: Thirty contact lens storage cases of 16 asymptomatic refractive surgery candidates were studied. Samples from the lens storage case fluid were inoculated into Bactec Peds Plus F broth (Becton Dickinson) and also directly onto blood agar, chocolate agar, and Sabouraud dextrose agar ("traditional method"). Another sample was processed for amoebal contamination. The rate of contamination of cases and the types of pathogens were evaluated for the broth-based and traditional culturing methods. Correlation between right and left storage cases of the same patient was defined as contamination of the two compartments with the same pathogen or pathogens. To avoid statistical bias, only one compartment was included in the study for these patients. RESULTS: Four storage cases were omitted due to growth correlation between right and left storage cases. Of the remaining 26 storage cases, 16 (61.5%) were found to be contaminated using the broth-based method and 10 (38.5%) using the traditional method (P=.011). High contamination rates were observed regardless of the type of disinfecting solution or type of contact lens used. CONCLUSIONS: The broth-based method had significantly greater culture yield than the traditional method. The high rates of contamination of contact lens storage cases of refractive surgery patients may put this cohort at greater risk than non-contact lens wearing candidates for developing postoperative infections.

Prediction potential of IR-micro spectroscopy for colon cancer relapse.

This study aimed to determine the potential of IR-spectroscopy to diagnose abnormality in histologically normal resection margins for predicting relapse in colon cancer patients. The present study evaluates potential abnormal crypt proliferation in histologically normal resection margins. Resection margins of 10 colon cancer patients (adenocarcinoma) (27 biopsies in all), found completely normal by standard histology were re-evaluated. Fourier transform infrared microscopy (FTIR-MSP) was performed on the longitudinal sections of the crypt, and spectral data collected from the base, middle and top portion of crypts. Absorbance in the region 900-1185 cm(-1) arising from carbohydrates and nucleic acids was found to be the most effective variate for such evaluation. In total 225 crypts were classified after assessing the levels of abnormality observed by the above technique. The abnormal biopsies detected using the above optical method was correlated with a relapse in the patient's history. Patients who had a relapse had at least one abnormal biopsy (crypt) based on the present methodology. Among the patients, the only case without a relapse was also the case where no abnormal crypts were found in any biopsies from the resection margins. The agreement between the biopsy status, as determined by the optical methodology, and the relapse of colonic malignancy based on the patients' medical files, establishes the translational nature of FTIR-MSP for medical purposes and hints at future clinical evaluation of the biopsies using this technique to determine more precisely the zone of excision during anastomosis.

Monitoring of viral cancer progression using FTIR microscopy: a comparative study of intact cells and tissues.

Fourier transform infrared microspectroscopy (FTIR-MSP) is an analytical method with a promising potential for detecting the spectral changes due to cancerous changes in cells. The purpose of the present study is monitoring biochemical spectral changes accompanying viral cancer progression in cells and tissues using FTIR-MSP. As a model system, we used cells in culture which were transformed to malignant cells by infection with murine sarcoma virus (MuSV) and cervical tissues at different neoplastic stages. In order to devise a systematic follow-up of the cancer progression, it was essential first to determine and validate consistent and significant spectral biomarkers, which can evidently discriminate between normal and cancerous cells/tissues. Then these biomarkers were used for the characterization and classification of early stages of malignant transformation utilizing discriminant classification function techniques. Our study points out that malignancy progression can be eminently graded for both cell lines and tissues. For example, using the array of four biomarkers: A(2958)A(2852)+A(2923),A(1121)/A(1015),A(1171)/A(1152)and|A(1082)-A(1056)|A(1028), we attained that the classification accuracies of different premalignant stages of cell lines and tissues were varied between 89.5 and 97.4%. These results strongly support the potential of developing FTIR microspectroscopy as a simple, reagent free method for early detection and accurate differentiation of premalignant stages.

Immune phenomena involved in the in vivo regression of fibrosarcoma cells expressing cell-associated IL-1alpha.

Constitutive expression of cell-associated, but not secreted, interleukin-1alpha (IL-1alpha) by oncogene-transformed fibrosarcoma cells induced regressing tumors in mice, a phenomenon that was abrogated by the IL-1 inhibitor, the IL-1 receptor antagonist (IL-1Ra). On the contrary, non-IL-1alpha-expressing tumor cells induce progressive tumors in mice. In vivo and ex vivo experiments have shown that regression of IL-1alpha-positive fibrosarcoma cells depends on CD8(+) T cells, which can also be activated in CD4(+) T cell-depleted mice, with some contribution of natural killer cells. In spleens of mice bearing the non-IL-1alpha-expressing fibrosarcoma cells, some early and transient manifestations of antitumor-specific immunity, such as activation of specific proliferating T cells, are evident; however, no development of cytolytic T lymphocytes or other antitumor protective cells could be detected. In spleens of mice bearing the non-IL-1alpha-expressing fibrosarcoma cells, the development of early tumor-mediated suppression was observed, and in spleens of mice injected with IL-1alpha-positive fibrosarcoma cells, protective immunity developed in parallel to tumor regression. Treatment of mice bearing violent fibrosarcoma tumors with syngeneic-inactivated, IL-1alpha-positive fibrosarcoma cells, at a critical interval after injection of the malignant cells (Days 5-12), induced tumor regression, possibly by potentiating and amplifying transient antitumor cell immune responses or by ablation of tumor-mediated suppression. Membrane-associated IL-1alpha may thus serve as an adhesion molecule, which allows efficient cell-to-cell interactions between the malignant and immune effector cells that bear IL-1Rs and function as a focused cytokine with adjuvant activities at nontoxic, low levels of expression. Our results also point to the potential of using antitumor immunotherapeutic approaches using cell-associated IL-1alpha.

Characterization of malignant melanoma using vibrational spectroscopy.

Malignant melanoma, a malignant neoplasm of epidermal melanocytes is the third most common skin cancer. In many cases, melanoma develops from nevus, which is considered as the nonmalignant stage. Fourier transform infrared microspectroscopy (FTIR-MSP), which is based on characteristic molecular vibrational spectra of cells, was used to investigate spectral differences between melanoma, nevus, and the corresponding normal epidermis. In the present work, FTIR-MSP was performed on formalin-fixed biopsies of melanoma and nevi along with the adjoining histologically normal epidermis to understand the biochemical variations from the epidermis and identify suitable parameters for differentiation of nevi from melanoma. The comparative analysis of various parameters calculated from the spectral data of the normal epidermis and the abnormal regions showed that the changes in the nucleic acids was a significant indicator of the abnormal nature of the tissues. The RNA/DNA ratio was decreased in case of both melanoma and nevus compared to the epidermis. The amide II/amide I ratio was greater for nevus and melanoma compared to the epidermis. In contrast to other organs, the analysis of carbohydrates was not found as a suitable indicator in case of malignant melanoma. Shifts in band wave number were found to be a major distinguishing feature between the melanoma and compound nevi. The present study helps in the identification of spectral features suitable for distinction of melanoma from nevus that appear similar even in FTIR spectral features and thus can pave the way for development of in vivo screening systems based on these diagnostic markers.

Inflammatory bowel diseases as an intermediate stage between normal and cancer: a FTIR-microspectroscopy approach.

Elucidation of the evolution of inflammatory bowel disease (IBD) to cancer by clinical symptoms and histopathology of biopsies is important. Fourier transform infrared microspectroscopy (FTIR-MSP) has shown promise as a diagnostic tool for distinction of normal and cancer cells and tissues. In the present work, FTIR-MSP is used to evaluate IBD cases and to study the IR spectral characteristic with respect to cancer and normal tissues from formalin-fixed colonic biopsies from patients. Specific regions of the spectra were analyzed by statistical tools to study variations in metabolites that signified changes between the two pathological conditions: IBD and cancer. IBD tissues can be grouped with cancer or normal tissue using certain parameters such as phosphate content and RNA/DNA ratio as calculated from the spectra and show intermediate levels with regard to these metabolites. Further classification of the spectra by cluster analysis indicated which cases of Crohn's disease (3 of 10 cases) or ulcerative colitis (7 of 10 cases) were more likely to progress to cancer. The study exhibits that FTIR-MSP can detect gross biochemical changes in morphologically identical IBD and cancer tissues and suggest which cases of IBD may require further evaluation for carcinogenesis.

Interleukin-6 and interleukin-10 are expressed in organs of normal young and old mice.

Interleukin-6 (IL-6) is a pleiotropic inflammatory cytokine, also endowed with inflammation-inhibiting properties. The status of interleukin-10 (IL-10) as an anti-inflammatory cytokine is more solidly established. The roles of IL-6 and IL-10 in the context of organ physiology, and their possible modulation by the aging process, are not satisfactorily understood. The purpose of this work was to characterize organ IL-6 and IL-10 expression in different cellular compartments in mice, under steady-state and stress conditions. The former was evaluated by immunohistochemistry (IHC) and enzyme-linked immunosorbent assay (ELISA) analyses of organ lysates (LYS) (addressing the intracellular compartment), while the latter was assessed by ELISA analyses of organ-conditioned media (CM), obtained after 48 hrs of organ culturing (addressing the potential of cytokine secretion/diffusion). Under steady-state conditions, the overall level of IL-6 and IL-10 expression was relatively low in both age groups (exceptionally, IHC staining demonstrated an enhanced expression of these cytokines in the heart, skeletal muscle and brain of young mice). Much more elevated levels of IL-6 and IL-10 expression were demonstrated in organ CM, possibly emphasizing the role of these cytokines in the context of organ stress. This was most characteristically shown in the highly specialized organs (heart, skeletal muscle and kidney) and liver of old mice, as compared with the other lymphoreticular organs (lungs, spleen, small intestine) tested. Thus, IL-10 was markedly upregulated in the highly specialized organs, while IL-6 was considerably reduced in the lymphoreticular organs. In addition, aging appears to be associated with altered patterns of intracellular expression and secretion/diffusion potentials of IL-6 and IL-10 in the heart and skeletal muscle, as demonstrated by reduced IHC staining on one hand, and an increased detection in organ CM, on the other. These findings may contribute to a better understanding of the unique functions of organ IL-6 and IL-10 in various age groups, and suggest an important role in organ response to stress in old age.

Diagnostic potential of Fourier-transform infrared microspectroscopy and advanced computational methods in colon cancer patients.

Colon cancer is the third leading class of cancer causing increased mortality in developed countries. A polyp is one type of lesion observed in a majority of colon cancer patients. Here, we report a microscopic Fourier transform infrared (FTIR) study of normal, adenomatous polyp and malignant cells from biopsies of 24 patients. The goal of our study was to differentiate an adenomatous polyp from a malignant cell using FTIR microspectroscopy and artificial neural network (ANN) analysis. FTIR spectra and biological markers such as phosphate, RNA/DNA derived from spectra, were useful in identifying normal cells from abnormal ones that consisted of adenomatous polyp and malignant cells. However, the biological markers failed to differentiate between adenomatous polyp and malignant cases. By employing a combination of wavelet features and an ANN based classifier, we were able to classify the different cells as normal, adenomatous polyp and cancerous in a given tissue sample. The percentage of success of classification was 89%, 81%, and 83% for normal, adenomatous polyp, and malignant cells, respectively. A comparison of the method proposed with the pathological method is also discussed.

Different patterns of interleukin-1alpha and interleukin-1beta expression in organs of normal young and old mice.

The different physiological roles of interleukin-1alpha (IL-1alpha) and interleukin-1beta (IL-1beta) are not well understood, especially when considering the apparent overlap and redundancy of the two IL-1 molecules. Characterization of IL-1alpha and IL-1beta expression was performed in this study in organs from young and old mice, using immunohistochemistry and ELISA (enzyme-linked immunosorbent assay). The results indicate that organ IL-1alpha and IL-1beta display different patterns of expression: IL-1alpha is manifested more prominently in lymphoreticular organs (lungs, small intestine, spleen, liver), while IL-1beta is more evident in highly specialized and more vulnerable organs, which do not play a leading role in defense against infections and intoxication (heart, brain, skeletal muscle, kidney). This differential expression is more accentuated in old mice, possibly pointing to the special relevance of these cytokines to organ homeostasis in old age. These findings may shed new light on the physiological functions of IL-1alpha and IL-1beta, and may also lead to the development of improved therapeutic approaches, based on the specific manipulation of these cytokines.

Spectroscopic evidence for site-specific cellular activity in the tubular gland in human intestine.

The intestinal crypts contain mucus-secreting goblet cells in large numbers. In the tubular gland (crypt), the cells are generated at the bottom and end their life cycle at the top. Recently, FTIR microspectroscopy (FTIR-MC) has been applied in biology and medicine. The characterization of various cellular types using FTIR-MC and its subsequent application for the diagnosis of cancer is becoming a reality. In this report, we investigate the differential cellular activity in the normal tubular gland using FTIR-MC. Our results indicate that the absorbance for the cells in the bottom of the crypt is always higher than those in the upper portion. There are spectral pattern changes and frequency shifts for cells at the bottom and top sites of the normal crypt. Also, the comparison of a normal crypt with a malignant one has been made. This is the first spectroscopic evidence in the literature showing the difference in the cellular activity at different sites in the tubular gland. The reasons for our observations and their implications are discussed.